Type 2 diabetes mellitus BALB/c mice are more susceptible to granulomatous amoebic encephalitis: Immunohistochemical study.

Granulomatous amoebic encephalitis (GAE) is a chronic, difficult to resolve infection caused by amphizoic amoebae of the genus Acanthamoeba, which in most cases occurs in immunosuppressed persons or with chronic diseases such as diabetes. In this study, we describe the early events of A. culbertsoni infection of GAE in diabetic mice model. Diabetes was induced in male BALB/c mice, with a dose of streptozotocin (130 mg/kg). Healthy and diabetic mice were inoculated via intranasal with 1 × 106 trophozoites of A. culbertsoni. Then were sacrificed and fixed by perfusion at 24, 48, 72 and 96 h post-inoculation, the brains and nasopharyngeal meatus were processed to immunohistochemical analysis. Invasion of trophozoites in diabetic mice was significantly greater with respect to inoculated healthy mice. Trophozoites and scarce cysts were immunolocalized in respiratory epithelial adjacent bone tissue, olfactory nerve packets, Schwann cells and the epineurium base since early 24 h post-inoculation. After 48 h, trophozoites were observed in the respiratory epithelium, white matter of the brain, subcortical central cortex and nasopharyngeal associated lymphoid tissue (NALT). At 72 h, cysts and trophozoites were immunolocalized in the olfactory bulb with the presence of a low inflammatory infiltrate characterized by polymorphonuclear cells. Scarce amoebae were observed in the granular layer of the cerebellum without evidence of inflammation or tissue damage. No amoebas were observed at 96 h after inoculation, suggesting penetration to other tissues at this time. In line with this, no inflammatory infiltrate was observed in the surrounding tissues where the amoebae were immunolocalized, which could contribute to the rapid spread of infection, particularly in diabetic mice. All data suggest that trophozoites invade the tissues by separating the superficial cells, penetrating between the junctions without causing cytolytic effect in the adjacent cells and subsequently reaching the CNS, importantly, diabetes increases the susceptibility to amoebae infection, which could favor the GAE development.

Resurrection of Mycteronastes (Monogenoidea: Monocotylidae), with Description of Mycteronastes caalusi n. sp. from Olfactory Sacs of the Smalltooth Sawfish, Pristis pectinata (Pristiformes: Pristidae), in the Gulf of Mexico off Florida.

Mycteronastes Kearn and Beverley-Burton, 1990 (Monogenoidea: Monocotylidae: Merizocotylinae) was resurrected from subjective synonymy with Merizocotyle Cerfontaine, 1894 , and its diagnosis was emended to include monocotylids with a haptor lacking a central loculus and having 5 peripheral (2 bilateral pairs and an unpaired anteromedial loculus), 1 interhamular, and 17 marginal loculi. The 3 species of Mycteronastes accepted herein are parasitic within the olfactory sacs of rays and sawfishes: Mycteronastes icopae ( Beverley-Burton and Williams, 1989 ) Kearn and Beverley-Burton, 1990 (type species) from the giant shovelnose ray, Glaucostegus typus (Anonymous (Bennett)) (Glaucostegidae), in the southwestern Pacific Ocean; Mycteronastes undulatae Kearn and Beverley-Burton, 1990 from the undulate ray, Raja undulata Lacepède (Rajidae), in the northeastern Atlantic Ocean; and Mycteronastes caalusi n. sp. from the smalltooth sawfish, Pristis pectinata Latham (Pristidae), in the Gulf of Mexico. Mycteronastes caalusi is most easily differentiated from its congeners by the combination of having 2 median cephalic papillae, an oval haptor that is wider than the body proper and lacks a deeply scalloped margin, a comparatively large anteromedial peripheral loculus, an unsclerotized male copulatory organ that is wholly anterior to the vaginal pores, a relatively small distal portion of the uterus (ootype chamber) that is mostly anterior to the vaginae, and a delicate uterus. The present study is the first report of a monocotylid from the olfactory sacs of P. pectinata and the first record of a species of Mycteronastes from the Gulf of Mexico. Notes on the taxonomy and systematics of some species assigned to Calicotyle Diesing, 1850 (Monocotylidae: Calicotylinae) are included.

Spatial Distribution of Gyrodactylus salmonis (Monogenea) on the Body of Captive Fingerling Oncorhynchus mykiss, Including Attachment Within the Olfactory Chamber.

Gyrodactylus salmonis is a common ectoparasite on the fins and body of North American salmonids in fresh water. In this study, the spatial distribution of G. salmonis on 60 captive hatchery-reared rainbow trout, Oncorhynchus mykiss , is reported. The highest parasite densities occurred on 5 × 5-mm(2) sections of the dorsal fin followed by the trunk, other fins, and the olfactory chamber, with the lowest densities on the head. The finding of infections within the olfactory chamber of 93% of the fish was unexpected. One possibility is that such infections represented spillover from high-density infrapopulations that occur on the skin and fins. However, this possibility is unlikely, because worm densities at various sites along the body surface of infected fish did not correlate with densities within the olfactory chamber. The parasite conceivably enters the chamber either via water incurrent or by crawling in from the head and subsequently remaining at this site to feed and reproduce. Results from scanning electron microscopy are consistent with physical modification to the olfactory epithelium associated with the attachment/reattachment of the opisthaptor and epithelial grazing.

Gender-associated differential expression of cytokines in specific areas of the brain during helminth infection.

Intraperitoneal infection with Taenia crassiceps cysticerci in mice alters several behaviors, including sexual, aggressive, and cognitive function. Cytokines and their receptors are produced in the central nervous system (CNS) by specific neural cell lineages under physiological and pathological conditions, regulating such processes as neurotransmission. This study is aimed to determine the expression patterns of cytokines in various areas of the brain in normal and T. crassiceps-infected mice in both genders and correlate them with the pathology of the CNS and parasite counts. IL-4, IFN-γ, and TNF-α levels in the hippocampus and olfactory bulb increased significantly in infected male mice, but IL-6 was downregulated in these regions in female mice. IL-1ß expression in the hippocampus was unaffected by infection in either gender. Our novel findings demonstrate a clear gender-associated pattern of cytokine expression in specific areas of the brain in mammals that parasitic infection can alter. Thus, we hypothesize that intraperitoneal infection is sensed by the CNS of the host, wherein cytokines are important messengers in the host-parasite neuroimmunoendocrine network.

Targeting the olfactory bulb during experimental cerebral malaria.

Malaria is responsible for over 500 million clinical cases and over 500000 deaths annually. Fatalities arise from a range of overlapping syndromes, such as cerebral malaria, whose pathogenesis is still incompletely understood. In a new study, Coban and colleagues provide new clues on the involvement of the olfactory bulb during experimental cerebral malaria in mice that open the way to testable hypotheses and potentially earlier intervention in humans.

Olfactory plays a key role in spatiotemporal pathogenesis of cerebral malaria.

Cerebral malaria is a complication of Plasmodium falciparum infection characterized by sudden coma, death, or neurodisability. Studies using a mouse model of experimental cerebral malaria (ECM) have indicated that blood-brain barrier disruption and CD8 T cell recruitment contribute to disease, but the spatiotemporal mechanisms are poorly understood. We show by ultra-high-field MRI and multiphoton microscopy that the olfactory bulb is physically and functionally damaged (loss of smell) by Plasmodium parasites during ECM. The trabecular small capillaries comprising the olfactory bulb show parasite accumulation and cell occlusion followed by microbleeding, events associated with high fever and cytokine storm. Specifically, the olfactory upregulates chemokine CCL21, and loss or functional blockade of its receptors CCR7 and CXCR3 results in decreased CD8 T cell activation and recruitment, respectively, as well as prolonged survival. Thus, early detection of olfaction loss and blockade of pathological cell recruitment may offer potential therapeutic strategies for ECM.

Immunohistochemical characterization of the initial stages of Naegleria fowleri meningoencephalitis in mice.

The initial stages of Naegleria fowleri meningoencephalitis in mice were immunohistochemically characterized following the first 8 h post-intranasal inoculation. The events found after 8 h were: (1) amebas in contact with the mucous layer of the olfactory epithelium, (2) numerous parasites eliminated by extensive shedding of the mucous layer, and (3) many organisms reaching the nasal epithelium. In contrast to other works, we observed that after 24 h, amebas invaded the epithelium, without evidence of the disruption of the nasal mucosa. In addition some trophozoites invading through the respiratory epithelium were observed, suggesting an additional invasion route. The inflammatory response detected was scarce until 30 h post-inoculation. After 96 h, the inflammatory response was severe in the olfactory bulb and brain, and the tissue damage great. Consequently, an inflammatory reaction may enhance tissue damage but apparently does not destroy amebas which seem to proliferate in the olfactory bulb.

The role of blood vessels and lungs in the dissemination of Naegleria fowleri following intranasal inoculation in mice.

Primary amoebic meningoencephalitis (PAM) was induced in mice by intranasal inoculation of Naegleria fowleri (Singh et Das, 1970) to study the role of the blood vessels and lungs in the early and later stages in this disease. Upon culturing blood and lung tissue obtained at 24-, 36-, 48-, 72-, 96-, and 120-hour time periods, it was found that amoebae grew only from blood and lung tissue obtained at the 96 and 120 hour time periods. Paraffin sections of the head revealed small foci of acute inflammation and amoebae within the olfactory bulb of the central nervous system (CNS) at 24 hours. Amoebae were not observed within blood vessels of the CNS until 96 and 120 hours. Also, amoebae were observed within the connective tissue surrounding blood vessels and sutures of the skull, bone marrow, and venous sinusoids between the skull bone tables at 96 and 120 hours. No amoebae or acute inflammatory reactions were observed in the lung sections from any time period and indirect immunofluorescence microscopy was negative for N. fowleri. This study provides evidence that neither blood vessels nor lungs provide routes for N. fowleri to the CNS during the early stages of PAM and that amoebae enter veins of the CNS and bone marrow during later stages of the disease.

[Studies on chemotactic behaviors of Strongyloides ratti infective larvae in vitro and migration routes of the larvae in rat head].

For the purpose of clarifying host-finding mechanisms, the infective larvae of Strongyloides ratti were run on an agarose plate which had been placed various attractants around the larvae. They were accumulated at the sites of sodium compounds, serum proteins and albumins. At 20, 30, 40 hours after infection, rats were killed and heads were separated. They were then processed for thin section specimens. Larvae were found in brains, olfactory bulbs and its nerves. These larvae seemed to move along olfactory nerves during descending from the cranial cavity. Another migration study was done by whole body autoradiography after 35S-methionine labeling for clarifying the larval pathway how to invade cranial cavity in the rats. They first crawled under subcutaneous region of rat and then accumulated at the tip of the nose. Next, they began to ascend olfactory nerves. They were in cranial cavity up to 20 hours after infection and then descending same route to nasal cavity. I don't know why they aim the cranial cavity but they had necessity to invade the cranial cavity and stay there for appropriate time to be an adult in the intestine.

The influence of inoculum size and time post-infection on the number and position of Toxocara canis larvae recovered from the brains of outbred CD1 mice.

Outbred CD1 mice were administered doses of 1000 and 3000 Toxocara canis eggs and postmortem took place on days 7, 42 and 120 post-infection. Mice were killed by cervical dislocation and brains were sagitally bisected and fixed in 10% neutral buffered formalin prior to histological preparation and examination. The number of T. canis larvae were counted per brain and per section and the number of larvae cited for the first time per section were also recorded. These observations were compared by dose administered and by day of postmortem. The total number of larvae per brain and per section was higher for the 3000 dose compared to the 1000 dose. A different pattern emerged for the number of larvae observed in the brain over the three postmortem days depending upon the dose received. For the 1000 dose larval numbers increase from day 7 to day 120 whereas for the 3000 dose the opposite trend occurs. Larvae were assigned to one of five regions in the brain - the telencephalon, diencephalon, cerebellum, medulla, pons and brain stem and the olfactory bulb. Larvae did not show a random distribution in the brain. The majority of larvae were recorded from the telencephalon and the cerebellum. The percentage of sections with larvae in them is higher for the 3000 dose compared to the 1000 dose for all regions of the brain. For the majority of regions, the percentage of sections with larvae in them increases between day 7 and 42 and then decreases by day 120 and this is most pronounced for the cerebellum. For the telencephalon and diencephalon only, more larvae were detected on the right hand side of the brain compared to the left hand side. Statistical analysis revealed that dose and brain region are significant factors which influence the number of larvae observed in histological sections of the brain but day post-infection is not.

A light microscopy study of the migration of Naegleria fowleri from the nasal submucosa to the central nervous system during the early stage of primary amebic meningoencephalitis in mice.

The migratory pathway of Naegleria fowleri from the nasal submucosa to the central nervous system (CNS) during the early stage of primary amebic meningoencephalitis (PAM) was investigated in mice. Twenty-one-day-old CD-1 mice were inoculated by intranasal instillation of 1 x 10(6) amebas. Animals were divided into 3 groups of 5 and, after being anesthetized, were killed at intervals of 24, 32, and 48 hr postinoculation by transcardial perfusion with formaldehyde, acetic acid, and methanol. The heads were decalcified, divided in the midsagittal plane, and the area of the cribriform plate removed and embedded in paraffin. Serial sections were cut at 8 microm and stained with a combination of celestin blue, Harris' hematoxylin, and acid fuchsin for light microscopy. Focal inflammation and amebas were observed in the submucosal nerve plexus, olfactory nerves penetrating the cribriform plate, and the olfactory bulb of the brain as early as 24 hr postinoculation. The time periods selected assured that the disease process would not obliterate soft tissue structures. Earlier studies used moribund mice in which the inflammation and the number of amebas were overwhelming. The present study provides convincing evidence that amebas gain initial access to the CNS through olfactory nerves within the cribriform plate during the early stages of PAM.

Studies on the pathogenesis of Acanthamoeba-associated meningoencephalitis.

100 indoor swimming pools were examined for the presence of free-living amoebae. Limax amoebae of the genus Acanthamoeba could be isolated from 34 samples. Naegleria spp. were not present. After axenic cultivation the pathogenicity in mice was tested. Histological as well as immunohistological procedures were compared in order to identify the amoebae in brain sections and to clarify the route of infection. The results indicate a direct invasion of the central nervous system by acanthamoebae via the nasal mucosa and the olfactory bulb.