THE 72-KDA PROTEIN OF NAEGLERIA FOWLERI PLAYS AN IMPORTANT ROLE IN THE ADHESION OF TROPHOZOITES TO BALB/C MICE NASAL EPITHELIUM.

Naegleria fowleri is a protozoan that causes primary amebic meningoencephalitis (PAM). The infection occurs when the trophozoites enter the nasal cavity, adhere to the nasal mucosa, invade the epithelium, and migrate until they reach the olfactory bulb. Like other pathogens, there is evidence that the adhesion of N. fowleri to host cells is an important factor in the process of cytopathogenicity and disease progression. However, the factors involved in the adhesion of the pathogen to the cells of the nasal epithelium have not been characterized. The objective of this study was to identify a protein on the surface of N. fowleri, which could act as adhesin to the mouse nasal epithelium in the PAM model. The interaction between proteins of extracts of N. fowleri and cells of the nasal epithelium of BALB/c mice was analyzed using overlay and Western blot assays. A 72-kDa band of N. fowleri interacted directly with epithelial cell proteins, this polypeptide band was purified and analyzed by mass spectrometry. Analysis revealed that polypeptide bands of 72 kDa contained peptides that matched the membrane protein, actin 1 and 2, and Hsp70. Moreover, the N. fowleri extracts resolved in 2D-SDS-PAGE showed that 72-kDa spot interacted with proteins of mouse epithelial cells, which include characteristics of the theoretical data of molecular weight and pH obtained in the analysis by mass spectrometry. Immunofluorescence tests showed that this protein is located on the surface of trophozoites and plays an important role in the adhesion of amoeba either in vitro or in vivo assays, suggesting that this protein contributes during the N. fowleri invasion and migration to the brain, causing primary amoebic meningoencephalitis.

Leishmania donovani mucosal leishmaniasis in Malta.

We report a case of a 76-year-old British man living in Malta who presented with a 7-month history of recurrent epistaxis and an enlarging right nasal vestibular lesion. Of note, his medical history included rheumatoid arthritis for which he was on long-term methotrexate. Blood results were unremarkable other than a mild lymphopaenia. Despite the use of various antibiotics and intranasal steroids, the lesion failed to resolve. This was eventually biopsied, and the histological picture was that of mucosal leishmaniasis. Leishmania donovani complex was detected by PCR. The patient was treated with liposomal amphotericin B on alternate days for a total of 20 doses. The lesion was found to have healed well at follow-up and the patient denied any further episodes of epistaxis.

Case Report: Mucosal Leishmaniasis Presenting with Nasal Septum Perforation after Almost Thirty Years.

Mucosal leishmaniasis (ML) is associated with progressive tissue destruction and granuloma formation, often after a considerable period of latency from an initial cutaneous infection. We report a case of recurrent epistaxis of 3 years duration and nasopharyngeal obstruction in a woman with treated cutaneous leishmaniasis nearly 30 years before and with no further exposure to Leishmania. Computed tomography revealed nasal septal perforation and histopathology demonstrated chronic inflammation. Microscopy was negative for amastigotes, but molecular testing of nasal mucosa biopsy detected Leishmania (Viannia) braziliensis. The patient underwent 28 days of treatment with IV sodium stibogluconate and her symptoms improved significantly. Sixteen months after treatment, she continues to have episodic epistaxis and detectable parasite load in her nasal lesion. Although ML is known to take years to decades to develop, there are few reported cases in the literature of such a long latency period. This report highlights the importance of considering ML in the differential diagnosis of chronic epistaxis in countries where leishmaniasis is endemic or in immigrants from these countries, even when presentation occurs decades after leaving an endemic region.

Preliminary report of histopathology associated with infection with tongue worms in Australian dogs and cattle.

Tongue worms utilise herbivorous mammals as intermediate hosts and reside in the nasopharynx of carnivores as their definitive hosts. A recent study in south eastern Australia showed an unexpectedly high infection (67%) of wild dogs with these parasites. The present study aimed at determining the pathogenicity of the parasite in both definitive (dog) and intermediate (cattle) hosts by histopathology. The definitive host showed multifocal haemorrhage of the interstitium of the nasal mucosa, multifocal mucosal erosion, congestion and haemorrhage, with haemosiderin laden macrophages present in those foci and distortion and destruction of the nasal mucosa. Histopathologic examination of lymph nodes from an infected cow showed diffuse eosinophilic granulomatous necrotising lymphadenitis and perinodal panniculitis with intralesional parasitic remnants and comparatively large numbers of eosinophils. A large, ~300-500 µm diameter, area of necrosis was also observed in one lymph node. This is the first time a study has been undertaken in Australia to determine the pathogenicity of tongue worms in both their definitive and intermediate hosts. This is a preliminary study and to properly estimate the health impact of infection with this pathogenic parasites on Australian production and companion animals more studies are necessary.

Successful Treatment of Sinusitis by Acanthamoeba in a Pediatric Patient After Allogeneic Stem Cell Transplantation.

Acanthamoeba infections are rare and mostly occur in immunocompromised patients. Most of the reported cases after stem cell transplantation have been diagnosed postmortem. We present the case of a 3-year-old boy with chronic graft versus host disease post hematopoietic transplantation, who was successfully treated for Acanthamoeba.

Miasis nasal: informe de un caso y revisión del tema / Nasal myiasis: report of a case and literature review / Miíase nasal: informe de um caso e revisão do assunto

La miasis es la infección de los tejidos u órganos de animales y seres humanos por larvas de dípteros; puede afectar a individuos de cualquier grupo etario, pero es más común en pacientes de edad media o avanzada. La miasis nasal, una infección de las cavidades nasales y paranasales por dichas larvas, es común en los países tropicales y en vías de desarrollo. Los casos informados de miasis nasal han sido causados por varias especies diferentes, entre ellas: Lucilia sericata en Corea e Irán, Estro ovis en Argelia y Francia, Lucilia cuprina y Phaenicia sericata en Malasia, Cochliomyia hominivorax en Guayana Francesa, Drosophila melanogaster en Turquía, Eristalis tenax en Irán y Oestrus ovis en Israel. Los principales signos y síntomas están relacionados con la presencia y el movimiento de las larvas, e incluyen sensación de cuerpo extraño, sangrado o secreción nasal mucopurulenta. La prevención se puede hacer con repelentes de insectos. El tratamiento de la miasis nasal se basa en el uso de antiparasitarios y técnicas para la eliminación de las larvas, pero puede incluir el uso de antibióticos profilácticos, sean tópicos o sistémicos, para las posibles infecciones secundarias. Se presenta un caso de miasis nasal y del seno maxilar izquierdo en una mujer de edad avanzada, que evolucionó favorablemente con el tratamiento.

Myiasis is the infection of animal or human tissues or organs by larvae of Diptera. It may affect individuals of any age, but is more common in middle-aged and elderly patients. Nasal myiasis, an infection of the nasal and paranasal cavities by such larvae, is a common disease in tropical and developing countries. Reported cases of nasal myiasis have been caused by several different species, such as Lucilia sericata in Korea and Iran, Estro ovis in Algeria and France, Lucilia cuprina and Phaenicia sericata in Malaysia, Cochliomyia hominivorax in French Guiana, Drosophila melanogaster in Turkey, Eristalis tenax in Iran and Oestrus ovis in Israel. Signs and symptoms are related to the presence and movement of the larvae, and include foreign body sensation, bloody or muco-purulent nasal discharge. Prevention may be done with insect repellent. Treatment is based on antiparasitic drugs and techniques for removal of larvae, but may include the use of prophylactic topical or systemic antibiotics for possible secondary infections. We report a case of nasal and left maxillary sinus myiasis in an elderly woman, whoresponded favorably to treatment.

A miíase é a infecção dos tecidos ou órgãos de animais e seres humanos por larvas de dípteros; pode afetar a indivíduos de qualquer faixa etária, mas é mais comum em pacientes de meia idade ou avançada. A miíase nasal, uma infecção das cavidades nasais e paranasais por ditas larvas, é comum nos países tropicais e em via de desenvolvimento. Os casos informados de miíase nasal tem sido causados porvarias espécies diferentes, entre elas: Lucilia sericata na Coreia e no Iran, Estro ovis na Argélia e na França, Lucilia cuprina e Phaenicia sericata na Malásia, Cochliomyia hominivorax na Guayana Francesa, Drosophila melanogaster na Turquía, Eristalis tenax na Iran e Oestrus ovis em Israel. Os principais signos e sintomas estão relacionados com a presencia e o movimento das larvas, e incluem sensação de corpo estranho, sangrado ou secreção nasal mucopurulenta. A prevenção se pode fazer com repelentes de insetos. O tratamento da miíase nasal se baseia no uso de antiparasitários e técnicas para a eliminação das larvas, mas pode incluir o uso de antibióticos profilácticos, sejam tópicos ou sistémicos, para as possíveis infecções secundárias. Se apresenta um caso de miíase nasal e do seno maxilar esquerdo numa mulher de idade avançada, que evolucionou favoravelmente com o tratamento.

Correlation between presence of Leishmania RNA virus 1 and clinical characteristics of nasal mucosal leishmaniosis / Correlação entre a presença de Leishmania RNA Vírus 1 e as características clínicas da leishmaniose de mucosa nasal

ABSTRACT INTRODUCTION: Mucosal leishmaniosis (ML) is a severe clinical form of leishmaniosis. Complex factors related to the parasite and the host are attributed to the development of mucosal lesions. Leishmania RNA virus 1 (LRV1) can disrupt immune response, and may be the main determinant of severity of the disease; it should be investigated. OBJECTIVE: To study the existence of clinical differences between patients with ML with endosymbiosis by LRV1 and. those without it. METHODS: A cross-sectional cohort study with clinical evaluation, polymerase chain reaction (PCR) detection of Leishmania, species classification, and search of LRV1 was performed. Only patients with confirmed diagnosis of ML by positive PCR and with nasal mucosa injuries were included in this analysis. RESULTS: Out of 37 patients, 30 (81.1%) were diagnosed with Leishmania braziliensis, five (13.5%) with Leishmania guyanensis, and two (5.4%) with mixed infection of L. braziliensis and L. guyanensis. LVR1 virus was present in 26 (70.3%) of the cases. CONCLUSION: Correlation between clinical phenotype and presence of LRV1 was not observed, although the frequency of the virus is two-fold higher in mucosal lesions than that found in the literature on skin lesions in the same geographical area.

RESUMO Introdução: A leishmaniose de mucosa (LM) é uma forma clínica grave da leishmaniose. Fatores complexos ligados ao parasita e ao hospedeiro são atribuídos ao desenvolvimento das lesões de mucosa. Leishmania RNA Vírus 1 (LRV1) pode subverter a resposta imune, podendo ser o principal determinante da gravidade da doença e deve ser pesquisado. Objetivo: Estudar a existência de diferenças clínicas entre pacientes portadores de LM com endosimbiose por LRV1 e as que não possuem. Métodos: Foi realizado um estudo de coorte histórica com corte transversal com avaliação clínica, detecção da Leishmania por técnica de PCR, classificação da espécie e pesquisa de LRV1. Foram incluídos na análise da pesquisa somente os pacientes com diagnóstico confirmado de LM com PCR positivo, com lesão de mucosa nasal. Resultados: Dos 37 pacientes, 30 (81,1%) foram diagnosticados com L. braziliensis, 5 (13,5%) com L. guyanensis e 2 (5,4%) com infecção mista de L. braziliensis e L. guyanensis. O vírus LVR1 estava presente em 26 casos (70,3%). Conclusão: A correlação entre o fenótipo clínico e a presença do LRV1 não foi constatada, porém a frequência do vírus é duas vezes maior em lesão de mucosa do que encontrado em trabalho, da mesma região, sobre lesão cutânea.

Mucosal relapse of visceral leishmaniasis in a child treated with anti-TNFα.

Visceral leishmaniasis is an enzootic parasitosis present across the Mediterranean Basin. Some consider it an opportunistic parasite. We report the case of a girl treated with anti-tumour necrosis factor alpha (anti-TNFα) for juvenile idiopathic arthritis who had previously presented with visceral leishmaniasis. Two and a half years later, she presented a tumour-like mass in the nasal mucous membrane caused by Leishmania parasites. Leishmania infantum is classically responsible for visceral leishmaniasis, but pure mucocutaneous leishmaniasis has also been described. To our knowledge, this is the first observation of a recurrence of visceral leishmaniasis in the mucocutaneous form. The occurrence of atypical forms and presentations in those on anti-TNF therapy should be considered.

Genotyping of potentially pathogenic Acanthamoeba strains isolated from nasal swabs of healthy individuals in Peru.

Free Living Amoebae (FLA) of Acanthamoeba genus are widely distributed in the environment and can be found in the air, soil and water; and have also been isolated from air-conditioning units. In humans, they are causative agents of a sight-threating infection of the cornea, Acanthamoeba keratitis (AK) and a fatal infection of the central nervous system known as Granulomatous Amoebic Encephalitis (GAE). In this study, a survey was conducted in order to determine the presence and pathogenic potential of free-living amoebae of Acanthamoeba genus in nasal swabs from individuals in two regions of Peru. Identification of isolates was based on cyst morphology and PCR-sequencing of the Diagnostic Fragment 3 to identify strains at the genotype level. The pathogenic potential of the isolates was also assayed using temperature and osmotolerance assays and extracellular proteases zymograms. The obtained results revealed that all isolated strains exhibited pathogenic potential. After sequencing the highly variable DF3 (Diagnostic Fragment 3) region in the 18S rRNA gene as previously described, genotype T4 was found to be the most common one in the samples included in this study but also genotype T15 was identified. To the best of our knowledge, this is the first study on the characterization of Acanthamoeba strains at the genotype level and the first report of genotype T4 and T15 in Peru.

Molecular diagnosis and species identification of mucosal leishmaniasis in Iran and correlation with cytological findings.

OBJECTIVE: Mucosal leishmaniasis (ML) is a rare destructive disease that mainly affects the mucous membranes of the mouth and nose. The etiologic agent(s) of ML are not well known in the Middle East. STUDY DESIGN: Cytologic smears of ML from the mucosal lesions of 7 patients were prepared by scraping. In 2 patients with nasal lesions, exfoliative cytology was made by washing the nasal cavity. The smears were both air dried and fixed in alcohol and stained. Scrapings from the same smears were then tested for leishmanial DNA by nested PCR. RESULTS: This study characterized 9 isolates of ML, with 7 cases identified as Leishmania major and 2 as Leishmania tropica. While 6 patients were found to be positive by the cytology technique, the nested PCR was positive for all of these samples. CONCLUSIONS: Presence of granuloma and multinucleated giant cells in the negative smears of the patients who showed clinical manifestation of ML was an important clue for diagnosis of this disease. The PCR-based method not only appears to be a precise diagnostic approach in the identification of suspected cases of ML but is also efficient in determining the species of the parasite. L. major and L. tropica can lead to ML, but they result in different cytologic features.

Anti-tumor necrosis factor-alpha therapy provokes latent leishmaniasis in a patient with rheumatoid arthritis.

It has been reported that anti-tumor necrosis factor-alpha therapy increases the risk of opportunistic infections including rare case reports of leishmaniasis. Here we report a case of latent cutaneous leishmaniasis, which was provoked by anti-tumor necrosis factor-alpha therapy in a patient with rheumatoid arthritis.

[Primary Leishmania infantum MON-80 endonasal leishmaniasis in Tunisia]. / Leishmaniose endonasale primitive à Leishmania infantum MON-80 en Tunisie.

BACKGROUND: Mucocutaneous leishmaniasis is endemic in Central and South America. It causes massive mutilating and disfiguring lesions and can lead to destruction of facial structures. In Tunisia, leishmaniasis of the mucous membranes is rare, usually developing as a complication of cutaneous leishmaniasis via direct extension. We report the first case in Tunisia of isolated and primary nasal leishmaniasis. CASE REPORT: A 70-year-old man with a history of nephrectomy for renal lithiasis was seen with a painless nodule that had been present for one month. The latter was erythematous, polypoid and firm, with a diameter of 2 cm, and was situated in the right endonasal mucosa. The diagnosis of leishmaniasis was confirmed by histological and direct examinations revealing high numbers of amastigotes of Leishmania. Culture of the offending organism in NNN medium and isoenzymatic characterization resulted in identification of MON-80 Leishmania infantum leishmaniasis. The outcome was good with treatment, and the nodule was deflated after six months. DISCUSSION: There have been few reports of similar cases of primary and isolated mucosal leishmaniasis caused by Leishmania infantum. Our case is also unusual in that zymodeme MON-80 is only rarely a cause of Mediterranean leishmaniasis.

Intranasal immunization with Naegleria fowleri lysates and Cry1Ac induces metaplasia in the olfactory epithelium and increases IgA secretion.

According to previous reports, intranasal administration of the Cry1Ac protein alone or with amoebic lysates increases protection against Naegleria fowleri meningoencephalitis in mice, apparently by eliciting IgA responses in the nasal mucosa. In the current study, we performed an immunohistochemical analysis of IgA in the nasal mucosa of mice immunized intranasally with Cry1Ac, and amoebic lysates or a combination of both. The animals were sacrificed 24 h after the last immunization or after an intranasal lethal challenge with N. fowleri. Our results indicate that all of the intranasal immunizations provoked an increase in areas with metaplasia in the olfactory epithelium, allowing for secretion of IgA. As a result, IgA antibodies were found interacting with trophozoites in the nasal lumen, and there was a marked increase of IgA in the metaplasic epithelium. On the other hand in nonimmunized mice trophozoites were observed invading the nasal mucosa, which was not the case for immunized mice. Our results suggest that intranasal immunization provokes cellular changes in the olfactory epithelium, leading to greater protection against N. fowleri that is probably caused by an increased secretion of IgA. The increased IgA response induced in the nasal mucosa by immunization probably impedes both amoebic adhesion and subsequent invasion of the parasite to the nasal epithelium.

Terminal phase of bird schistosomiasis caused by Trichobilharzia regenti (Schistosomatidae) in ducks (Anas platyrhynchos f. domestica).

The terminal phase of the migration of Trichobilharzia regenti Horák, Kolárová et Dvorák, 1998 in the definitive host (Anas platyrhynchos f. domestica) was studied 12-27 days post infection (p.i.). Brain meninges were the last part of the nervous system where the worms were detected before their occurrence in the nasal cavity. In meninges, the parasites started to feed on red blood cells. Then the worms occurred in the nasal mucosa 14-25 days p.i. and the first immature eggs appeared 15 days p.i. The fully developed miracidia were recorded in the eggs from 17 days p.i. and freely in the nasal mucosa 19 days p.i. Infiltrates of lymphocytes, later also eosinophils and heterophils around the eggs and free miracidia, were observed from 15 and 19 days p.i., respectively. The haemorrhages occurring from 17 days p.i., and the granulomas with lymphocytes, eosinophils and heterophils forming around the eggs from 22 days p.i. were the most apparent pathological changes of nasal tissue.

Leishmaniasis of the nasal cavity: a case report.

Establishing diagnosis of a granulomatous lesion of the nose is often difficult. Here we report a case of granulomatous lesion of the nose caused by Leishmania--an unlikely cause in the UK. The diagnosis and management of the case is discussed here.

Leishmania (Viannia) braziliensis-induced chronic granulomatous cutaneous lesions affecting the nasal mucosa in the rhesus monkey (Macaca mulatta) model.

The present studies on infections with Leishmania (Viannia) braziliensis in rhesus macaques were made to characterize the evolution of different parasite strains and the immune responses they elicited in this experimental host. A standardized inoculum of promastigotes was injected intradermally either above the eyelid or on the forearm of each monkey. Sixteen infected monkeys developed longstanding infections which lasted until the end of the observation period (33 months). The time required for lesion development was very variable, not only for the isolates showing molecular differences but also for individual animals in groups infected with the same parasite strain. The inocula produced lesions of variable severity, ranging from localized cutaneous leishmaniasis (CL) with a tendency to spontaneous healing to non-healing disease. One infected animal developed persistent metastatic skin and mucosal lesions. Anti-Leishmania antibodies and parasite-specific T-cell responses were induced by the experimental infections. As the granulomatous inflammatory response found at the lesions in L. (V.) braziliensis-infected M. mulatta was similar to that in patients with CL, this primate model could be useful for studying the pathophysiology and immunoregulatory events associated with disease evolution, as well as for the evaluation of new drugs or candidate vaccines.

Biological behavior of Leishmania (L.) amazonensis isolated from a human diffuse cutaneous leishmaniasis in inbred strains of mice.

After a subcutaneous injection of 100000 purified amastigotes of an isolate from a diffuse case of cutaneous leishmaniasis caused by the MHOM/BR/76/Ma-5 strain of Leishmania amazonensis, three inbred mouse strains developed a progressive nodular lesion, which evolved to an ulcerated lesion. Based on these data, mice of BALB/c, C57BL/6 or C57BL/10 could be classified as susceptible. The majority of mice developed metastases in the footpads, ear, tail, nose and oral mucosa. Amputation of the members related to the primary lesion was frequent. Experiments using the limiting dilution analysis showed that there was no correlation between lesion and parasite load. It has been demonstrated that these mouse strains could be considered excellent models for mucocutaneous leishmaniasis when infected with L. amazonensis. Metastatic lesions caused destruction of the nasal region with many parasitized macrophages under the epithelial surface of the nasal mucosa. Bone destruction occurred with an extensive inflammatory reaction presenting macrophages heavily parasitized by amastigotes. The parasites also spread to the periodontal ligament and other structures of the oral cavity, which could induce a severe inflammatory process. This study indicates that both nasal and oral lesions in mice infected by L. amazonensis were characterized by an inflammatory reaction with the presence of a high parasite load within macrophages.

[Disseminated microspiridiosis (Encephalitozoon intestinalis) in a patient with HIV infection]. / Disseminierte Mikrosporidieninfektion (Encephalitozoon intestinalis) bei HIV-Infektion.

HISTORY AND CLINICAL FINDINGS: A 39-year-old patient with advanced HIV infection was admitted to our hospital with a 6-month history of diarrhoea, abdominal pain and pansinusitis. INVESTIGATIONS: Ultrasound and endoscopic retrograde cholangiography revealed cholangitis of the larger bile ducts. Stool examinations and coloscopy were unremarkable. No pathogenic organisms were identified by routine investigations. Finally, microsporidia of the genus encephalitozoon were diagnosed by electron microscopy in biopsies from the bile duct and the nasal mucous membrane and in stool samples by polymerase chain reaction (PCR). TREATMENT AND COURSE: Albendazole treatment was successful. The cholestatic liver tests and the ultrasound findings normalized. Control tests of stool, bile and nasal secretions by light microscopy, electron microscopy, and PCR were negative. CONCLUSION: Microsporidia, along with human cytomegalovirus, cryptosporida and mycobacteria other than tuberculosis are increasingly recognized as causing opportunistic infections in immunodeficient patients, especially in AIDS-related cholangitis. Some species can cause systemic infection. Therefore microsporidia infection should be considered in the differential diagnosis of all patients with immunodeficiency.

Systemic leishmaniasis involving the nose.

A case of a patient, presenting with a granulomatous lesion of the anterior nasal septum mucosa spreading to the columella and the nasal floor, whereby leishmaniasis was diagnosed, is presented. The clinical and pathological aspects of this pathology, its diagnosis and treatment are reviewed.

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.