SummaryOrganisms of the Mycobacterium avium complex (MAC) are widely distributed in the environment, form biofilms in water pipes and potable water tanks, and cause chronic lung infections in patients with chronic obstructive pulmonary disease and cystic fibrosis. Pathological studies in patients with pulmonary MAC infection revealed granulomatous inflammation around bronchi and bronchioles. BEAS-2B human bronchial epithelial cell line was used to study MAC invasion. MAC strain A5 entered polarized BEAS-2B cells with an efficiency of 0.1 ± ± ± ± 0.03% in 2 h and 11.3 ± ± ± ± 4.0% in 24 h. In contrast, biofilm-deficient transposon mutants 5G4, 6H9 and 9B5 showed impaired invasion. Bacteria exposed to BEAS-2B cells for 24 h had greater ability to invade BEAS-2B cells compared with bacteria incubated in broth. M. avium had no impact on the monolayer transmembrane resistance. Scanning electron microscopy showed that MAC A5 forms aggregates on the surface of BEAS-2B cell monolayers, and transmission electron microscopy evidenced MAC within vacuoles in BEAS-2B cells. Cells infected with the 5G4 mutant, however, showed significantly fewer bacteria and no aggregates on the cell surface. Mutants had impaired ability to cause infection in mice, as well. The ability to form biofilm appeared to be associated with the invasiveness of MAC A5.

Domestic dogs and cats are commonly infected with a variety of protozoan enteric parasites, including Blastocystis spp. In addition, there is growing interest in Blastocystis as a potential enteric pathogen, and the possible role of domestic and in-contact animals as reservoirs for human infection. Domestic animals in shelter environments are commonly recognized to be at higher risk for carriage of enteropathogens. The purpose of this study was to determine the frequency of infection of shelter-resident and client-owned domestic dogs and cats with Blastocystis spp in the Pacific Northwest region of the USA. Fecal samples were collected from 103 shelter-resident dogs, 105 shelter-resident cats, 51 client-owned dogs and 52 client-owned cats. Blastocystis were detected and subtypes assigned using a nested PCR based on small subunit ribosomal DNA sequences. Shelter-resident animals were significantly more likely to test positive for Blastocystis (P<0.05 for dogs, P = 0.009 for cats). Sequence analysis indicated that shelter-resident animals were carrying a variety of Blastocystis subtypes. No relationship was seen between Blastocystis carriage and the presence of gastrointestinal disease signs in either dogs or cats. These data suggest that, as previously reported for other enteric pathogens, shelter-resident companion animals are a higher risk for carriage of Blastocystis spp. The lack of relationship between Blastocystis carriage and intestinal disease in shelter-resident animals suggests that this organism is unlikely to be a major enteric pathogen in these species.

The ability to infect macrophages is a common characteristic shared among many mycobacterial species. Mycobacterium avium, Mycobacterium tuberculosis, and Mycobacterium kansasii enter macrophages, using the complement receptors CR1, CR3, CR4, and the mannose receptor. To identify M. avium genes and host cell pathways involved in the bacterial uptake by macrophages, we screened a M. avium transposon mutant library for the inability to enter macrophages. Uptake-impaired clones were selected. Sequence of six M. avium clones identified one gene involved in glycopeptidolipid biosynthesis, one gene encoding the conserved membrane protein homologue to the M. avium subsp. paratuberculosis MAP2446c gene and four others belonging to the same region of the chromosome. Analysis of the chromosome region revealed a pathogenicity island inserted between two tRNA sequences with 58% of G؉C content versus 69% in the M. avium genome. The region is unique for M. avium and is not present in M. tuberculosis or M. paratuberculosis. Although the mutants did not differ from the WT bacterium regarding the binding to macrophage cell membrane, analysis of macrophage proteins after 1 h infection revealed a deficiency in the mutant to phosphorylate certain proteins on uptake. To understand M. avium interaction with two evolutionarily distinct hosts, the mutants were evaluated for Acanthamoeba castellanii invasion. The defect in the ability of the mutants to invade both cells was highly similar, suggesting that M. avium might have evolved mechanisms that are used to enter amoebas and human macrophages. uptake M ycobacterium avium complex is an intracellular pathogen that can infect a variety of host cells (1-4). M. avium, like the majority of pathogenic mycobacteria, infects and replicates within macrophages (5, 6), which are the phagocytic cells where M. avium establishes a long-term infection (7).A number of studies in vitro have determined that M. avium, in a manner similar to Mycobacterium tuberculosis, is taken up by macrophages using the complement receptors CR3, CR4, and CR1 (8-10), and/or the mannose receptor (8, 11). M. tuberculosis has been found to bind to the CD11b chain of the ␤-integrin of the CR3 and CR4 receptors, which happened without participation of the serum complement proteins (10). Other studies demonstrated that the mannose capped lipoarabinomannan antigen of virulent M. tuberculosis cell wall is capable to recognize and bind to the mannose receptors on the macrophage membrane (11).It has been hypothesized that virulent mycobacteria bind to many receptors because pathogenic mycobacteria would use several different receptors on the macrophage membrane to be internalized, as a strategy to guarantee phagocytosis, even when a sitespecific macrophage would not contain one or more membrane receptors. In addition, this property would address the possibility that mycobacteria may not express all of the ligands during all phases of infection. Most mycobacteria evolved to survive inside phagocytic cells, although they can enter ...

The centrifugation-flotation technique appeared to offer clear advantages in detecting infection with E macusaniensis, Trichuris spp, Nematodirus spp, and capillarids. The saline McMaster technique appeared to offer an advantage in detecting small coccidia.

BackgroundThe objective was to determine the effects of agility exercise on dogs of different skill levels with respect to urinary eicosanoids, urinary 15F2t-isoprostane (lipid peroxidation marker) and hematological/biochemical changes in plasma. Fifteen adult dogs had blood and urine samples obtained prior to, immediately and 4-hours following an agility exercise.ResultsHematocrit, red blood cells (RBC), albumin, and hemoglobin increased following exercise, with greatest increases correlating to increased skill group (novice, intermediate, masters); at 4-hours post-exercise, hematocrit, RBC, and hemoglobin were decreased. Phosphorus increased following exercise with the greatest increase in novice and intermediates. Plasma lactate increased 3.6-fold in masters, 3.2-fold in intermediates, and 1.2-fold in novice dogs. Urine thromboxane B2 (TXB2) more than tripled 4-hours post-exercise while 6-keto prostaglandin F1α (PGF1α, prostacyclin metabolite), prostaglandin E2 metabolites (13,14-dihydro-15-keto-prostaglandin A2 and 13,14-dihydro-15-keto-prostaglandin E2), and 13,14-dihydro-15-keto prostaglandin F2α were unaffected as determined by a competitive enzyme immunoassay and standardized by division with urine creatinine. Urine 15F2t-isoprostane increased insignificantly.ConclusionsAlterations in the plasma post-exercise were likely due to hemoconcentration from insensible water loss, splenic contraction and sympathetic stimulation while 4-hours later autohemodilution reduced RBC parameters. Elevations in plasma lactate and urinary TXB2 correlated with advanced skill level/speed of the dogs.