Encephalitozoon cuniculi is an important microsporidian pathogen that is considered an emergent, zoonotic, and opportunistic. It infects both domestic and laboratory rabbits, generating severe chronic interstitial and granulomatous nephritis with fibrosis and granulomatous encephalitis. Encephalitozoonosis is diagnosed in paraffin-embedded sections by examining the spores in the host tissues. The spores are difficult to observe when the samples are stained with hematoxylin and eosin (H&E), particularly when there is an inflammatory reaction and tissue damage. The spores are easily mistaken for other microorganisms, such as fungi (yeasts), protozoa, and bacteria. In our study, we used kidney samples from E. cuniculi–positive rabbits and employed 14 recommended histologic stains for detecting microsporidia spores: alcian blue, calcofluor white, Giemsa, Gram, Grocott, H&E, Luna, Luxol fast blue, Masson trichrome, modified trichrome stain (MTS), periodic acid–Schiff reaction (PAS), Van Gieson, Warthin–Starry (WS), and Ziehl–Neelsen (ZN).We concluded that MTS and Gram stain, detected by light microscopy, and calcofluor white stain, detected by ultraviolet light microscopy, are the best stains for detecting spores of E. cuniculi in paraffin-embedded tissues from infected rabbits. These stains were superior to WS, ZN, Giemsa, and PAS for identifying spores without background “noise” or monochromatic interference. Also, they allow individual spores to be discerned in paraffin-embedded tissues. MTS allows observation of the polar tube, polaroplast, and posterior vacuole, the most distinctive parts of the spore.

Helicobacter pyloriis a spiral Gram-negative bacterium associated with inflammation of the gastric mucosa, peptic ulcer, and gastric adenocarcinoma, whose treatment has failed due to antibiotic resistance and side effects. Furthermore, because there are no vaccines effective againstH. pylori, an appropriate vaccine design targeting conserved/essential genes must be identified. In the present study, aH. pylori50–52 kDa immunogen-derived peptide antigen with the sequence Met-Val-Thr-Leu-Ile-Asn-Asn-Glu (MVTLINNE) was used to immunize againstH. pyloriinfection. For this, mice received an intraperitoneal injection of 100 μg ofH. pyloripeptide on the first week, followed by two weekly subcutaneous reinforcements and further 10⁹bacteria administration in the drinking water for 3 weeks. Thymic cells proliferative responses to concanavalin A, serum levels of IL-2, IL-4, IL-6, IL-10, IL-17, IFN-γ, and TNF-αcytokines, and IgG1, IgG2a, IgG2b, IgG3 IgM, and IgA immunoglobulins were evaluated. Significant (p<0.05) increases on lymphoproliferation and spleen weights after immunization were observed. In contrast, infection significantly (p<0.05) decreased lymphoproliferation, which was recovered in immunized mice. In addition, levels of serum TH1 and TH2 cytokines were not altered after immunization, except for the significant increase in IL-6 production in immunized and/or infected animals. Moreover, immunization correlated with plasma secretory IgA and IgG, whereas infection alone only elicited IgM antibodies. Peptide immunization protected 100% of mice against virulentH. pylori. MVTLINNE peptide deserves further research as an approach to the prophylaxis ofH. pyloriinfection.

A female coyote (Canis latrans) was fatally injured by a vehicle on a road in San Luis Potosi, Mexico. Because of deteriorating clinical signs, the animal was euthanized. Postmortem examination of the lungs showed numerous small multifocal white nodules (0.5–1 cm diameter) disseminated throughout. Histopathologic examination revealed multifocal coalescing granulomas with abundant macrophages, numerous neutrophils, fibroblasts, plasma cells, and lymphocytes. Abundant intracellular and extracellular thick-walled, refractile, spherical yeasts (10–15 μm) were observed within the granulomas. The yeasts were intensely PAS-positive, with granular protoplasm. Broad-based single budding yeasts were occasionally present. Based on the microscopic findings of the pulmonary lesions and the morphological features of the organism, a diagnosis of chronic pyogranulomatous pneumonia caused byBlastomyces dermatitidiswas made. To our knowledge, the case described herein is the first report of pulmonary blastomycosis in a wild coyote.