Heme oxygenase-1 modulates brain inflammation and apoptosis in mice with angiostrongyliasis.

The rat nematode lungworm Angiostrongylus cantonensis undergoes obligatory intracerebral migration in its hosts and causes eosinophilic meningitis or meningoencephalitis. Heme oxygenase 1 (HO-1) has several cytoprotective properties such as anti-oxidative, anti-inflammatory, and anti-apoptotic effects. HO-1 in brain tissues was induced in A. cantonensis-infected group and showed positive modulation in cobalt protoporphyrin (CoPP)-treated groups. Assay methods for the therapeutic effect include western blot analysis, enzyme-linked immunosorbent assay, gelatin zymography, blood-brain barrier permeability evaluation and eosinophil count in cerebrospinal fluid. The combination of albendazole (ABZ) and CoPP significantly decreased pro-inflammatory cytokines, tumor necrosis factor-α, interleukin (IL)-1ß, IL-5, and IL-33 but significantly increased anti-inflammatory cytokines IL-10 and transforming growth factor-ß. In addition, worm recovery, matrix metalloproteinase-9, BBB permeability, and eosinophil counts were decreased in the ABZ and CoPP co-treated groups. Induction of HO-1 with CoPP strongly inhibited the protein levels of caspase-3 and increased the induction of annexin-V and B-cell leukemia 2. Thus, co-treatment with ABZ and CoPP prevented A. cantonensis-induced eosinophilic meningoencephalitis and its anti-apoptotic effect by promoting HO-1 signaling prior to BBB dysfunction. HO-1 induction might be a therapeutic modality for eosinophilic meningoencephalitis.

Exosome-Depleted Excretory-Secretory Products of the Fourth-Stage Larval Angiostrongylus cantonensis Promotes Alternative Activation of Macrophages Through Metabolic Reprogramming by the PI3K-Akt Pathway.

Angiostrongylus cantonensis (AC), which parasitizes in the brain of the non-permissive host, such as mouse and human, is an etiologic agent of eosinophilic meningitis. Excretory-secretory (ES) products play an important role in the interaction between parasites and hosts' immune responses. Inflammatory macrophages are responsible for eosinophilic meningitis induced by AC, and the soluble antigens of Angiostrongylus cantonensis fourth stage larva (AC L4), a mimic of dead AC L4, aggravate eosinophilic meningitis in AC-infected mice model via promoting alternative activation of macrophages. In this study, we investigated the key molecules in the ES products of AC L4 on macrophages and observed the relationship between metabolic reprogramming and the PI3K-Akt pathway. First, a co-culture system of macrophage and AC L4 was established to define the role of AC L4 ES products on macrophage polarization. Then, AC L4 exosome and exosome-depleted excretory-secretory products (exofree) were separated from AC L4 ES products using differential centrifugation, and their distinct roles on macrophage polarization were confirmed using qPCR and ELISA experiments. Moreover, AC L4 exofree induced alternative activation of macrophages, which is partially associated with metabolic reprogramming by the PI3K-Akt pathway. Next, lectin blot and deglycosylation assay were done, suggesting the key role of N-linked glycoproteins in exofree. Then, glycoproteomic analysis of exofree and RNA-seq analysis of exofree-treated macrophage were performed. Bi-layer PPI network analysis based on these results identified macrophage-related protein Hexa as a key molecule in inducing alternative activation of macrophages. Our results indicate a great value for research of helminth-derived immunoregulatory molecules, which might contribute to drug development for immune-related diseases.

Co-Therapy of Albendazole and Dexamethasone Reduces Pathological Changes in the Cerebral Parenchyma of Th-1 and Th-2 Dominant Mice Heavily Infected with Angiostrongylus cantonensis: Histopathological and RNA-seq Analyses.

Administration of albendazole alone was not very suitable for the treatment of cerebral angiostrongyliasis. This study was designed to evaluate the effects of the co-therapy of this drug and dexamethasone in Th-1 and Th-2 dominant mice infected with Angiostrongylus cantonensis. Each of BALB/c and C57BL/6 mice infected with 50 A. cantonensis third-stage larvae were administered albendazole (10 mg/kg/day) alone, dexamethasone (0.5 mg/kg/day) alone, or co-therapy of the two drugs from day 7 or 14 post-infection for 7 or 14 days. After sacrifice, coronal slices were prepared from five brain regions and stained with hematoxylin and eosin. Eight pathological changes were employed to determine the therapeutic effectiveness using a scoring system. RNA-seq analysis was performed to confirm the histopathological findings. The infected BALB/c and C57BL/6 mice had similar patterns in the pathological changes. Meningitis, hemorrhage, size of worms, and encephalitis in the cerebral parenchyma were slighter in the mice treated with co-therapy than the remaining groups. Mice treated from day 14 had more severe changes than those from day 7. The histopathological findings were found to be consistent to immune responses determined by RNA-seq analysis. Co-therapy was determined to reduce pathological changes after administration to mice infected with A. cantonensis.

Peroxisome-Proliferator Activator Receptor γ in Mouse Model with Meningoencephalitis Caused by Angiostrongylus cantonensis.

Peroxisome-proliferator activator receptor γ (PPARγ) has an anti-inflammatory role that inhibits the nuclear factor-κB (NF-κB) pathway and regulates the expressions of pro-inflammatory proteins, whereas its role in parasitic meningoencephalitis remains unknown. In this study we investigated the role of PPARγ and related mechanisms in eosinophilic meningoencephalitis caused by the rat lungworm Angiostrongylus cantonensis. We observed increased protein NF-κB expression in mouse brain tissue using GW9662, which is the specific antagonist of PPARγ, in a mouse model of angiostrongyliasis. Then we investigated NF-κB-related downstream proteins, such as COX-2, NOSs, and IL-1ß, with Western blot or enzyme-linked immunosorbent assay and found that the protein expression was upregulated. The results of gelatin zymography also showed that the MMP-9 activities were upregulated. Treatment with GW9662 increased the permeability of the blood-brain barrier and the number of eosinophils in cerebrospinal fluid. These results suggested that in angiostrongyliasis, PPARγ may play an anti-inflammation role in many inflammatory mediators, including NOS-related oxidative stress, cytokines, and matrix metalloproteinase cascade by decreasing the NF-κB action.

Lidocaine Infusion for Refractory Pain from Rat Lungworm Disease - Honolulu, Hawai'i.

Human infection with Angiostrongyloides cantonensis, or rat lungworm disease, manifests most commonly with neurologic symptoms that often include severe diffuse pain. While pain is reported by the majority of patients with rat lungworm disease, there are presently no published guidelines on the approach to pain management for these patients. Here we report a case of rat lungworm disease where severe refractory pain was the most prominent symptom and an intravenous lidocaine infusion was used as a successful treatment modality. Intravenous lidocaine has been shown to be safe and effective in neuropathic pain, refractory cancer pain, and peri-operative pain management. To our knowledge, this is the first case report on the use of lidocaine infusion for the management of refractory pain from rat lungworm disease, and among the first reports of any approach, to pain management for rat lungworm disease. We suggest that a lidocaine infusion protocol be considered when pain from rat lungworm disease fails to respond to first-line analgesics.

Angiostrongylus cantonensis causes cognitive impairments in heavily infected BALB/c and C57BL/6 mice.

BACKGROUND: Parasitic infections may cause significant effects on behavior, learning, and memory of the host. In the brain of mice heavily infected with Angiostrongylus cantonensis, severe damage has been observed in the hippocampus. This component has been considered to have associations with spatial learning and memory in humans and vertebrates. This study was designed to determine the impairments in behavior, learning, and memory in BALB/c and C57BL/6 mice heavily infected with the parasite. METHODS: Each mouse was inoculated with 50 third-stage larvae of A. cantonensis. After infection, daily changes in weight and dietary consumption, worm recoveries and survival rates were determined. The forced swimming test, open field test, and Morris water maze test were employed to evaluate depression- and anxiety-like behavior as well as impairments in spatial learning and memory, respectively. RESULTS: The worm recovery rate in the BALB/c mice was significantly lower than that of C57BL/6 mice from day 14 post-infection. The survival rate in infected BALB/c mice decreased to 0% by day 25 whereas those with swim-training survived three more days. On day 42, the C57BL/6 mice had a survival rate of 85.7% in the swimming group and 70% in the non-swimming group. Significant differences were found in weight between infected and non-infected BALB/c and C57BL/6 mice from day 13 and day 12, respectively with corresponding changes in their dietary consumption. Depression-like behavior was found in the infected BALB/c mice but not in C57BL/6 mice. However, anxiety-like behavior was found to occur only in C57BL/6 mice. Impaired spatial learning and memory were also found in the two strains of mice which occurred from day 14 post-infection. CONCLUSIONS: Results of this study indicate that A. cantonensis causes depression, anxiety, and impairments in spatial learning and memory in heavily infected mice. Moreover, significantly higher severity was observed in the Th-2 dominant BALB/c mice.

Excretory/secretory products of Angiostrongylus cantonensis fifth-stage larvae induce endoplasmic reticulum stress via the Sonic hedgehog pathway in mouse astrocytes.

BACKGROUND: Angiostrongylus cantonensis is an important food-borne zoonotic parasite. Humans are non-permissive hosts, and this parasite develops into fifth-stage larvae (L5) in the brain and subarachnoid cavity and then induces eosinophilic meningitis and eosinophilic meningoencephalitis. Excretory/secretory products (ESPs) are valuable targets for the investigation of host-parasite interactions. These products contain a wide range of molecules for penetrating defensive barriers and avoiding the immune response of the host. Endoplasmic reticulum (ER) stress has been found to be associated with a wide range of parasitic infections and inflammation. ER stress can increase cell survival via the activation of downstream signalling. However, the mechanisms of ER stress in A. cantonensis infection have not yet been clarified. This study was designed to investigate the molecular mechanisms of ER stress in astrocytes after treatment with the ESPs of A. cantonensis L5. RESULTS: The results demonstrated that A. cantonensis infection activated astrocytes in the mouse hippocampus and induced the expression of ER stress-related molecules. Next, the data showed that the expression of ER stress-related molecules and the Ca2+ concentration were significantly increased in activated astrocytes after treatment with the ESPs of L5 of A. cantonensis. Ultimately, we found that ESPs induced GRP78 expression via the Sonic hedgehog (Shh) signalling pathway. CONCLUSIONS: These findings suggest that in astrocytes, the ESPs of A. cantonensis L5 induce ER stress and that the Shh signalling pathway plays an important role in this process.

Particularidades de la meningoencefalitis eosinofílica en la era del caracol gigante africano / Particular features of eosinophilic meningoencephalitis in the age of the giant African snail

Introducción: la meningoencefalitis eosinofílica es una enfermedad inflamatoria infecciosa reportada en Cuba desde la década de los ochenta del siglo pasado y actualmente extendida al continente americano. Es producida por el parásito Angiostrongylus cantonensis. Objetivo: determinar si existen diferencias entre los pacientes que sufrieron meningoencefalitis eosinofílica antes y después de la introducción del caracol gigante africano. Métodos: se estudiaron un total de 19 muestras de líquido cefalorraquídeo y suero tomadas simultáneamente a cada paciente diagnosticados con meningoencefalitis eosinofílica, perteneciente a la seroraquioteca del Laboratorio Central del Líquido Cefalorraquídeo (LABCEL). Para la determinación de las proteínas albúmina e IgG se empleó como método de laboratorio la inmunodifusión radial. Resultados: el 14 por ciento de los pacientes fueron adultos antes de la aparición de este molusco, en contraste con el momento actual, donde el 50 por ciento son pacientes con edades superiores a 18 años. El porcentaje de síntesis de IgG intratecal media fue mayor en los enfermos actuales, aunque no de manera significativa. Conclusiones: . existen diferencias antes y después de la aparición del caracol gigante africano dado por la respuesta de síntesis intratecal en los pacientes asociados con el molusco que denota una mayor agresividad del parásito. La edad promedio mayor de los enfermos confirma que estos son los que más manipulan y dispersan el molusco(AU)

Introduction: eosinophilic meningoencephalitis is an infectious inflammatory disease reported in Cuba since the 1980s and currently extended to the American continent. This condition is caused by the parasite Angiostrongylus cantonensis. Objective: determine whether there are differences between the patients suffering from eosinophilic meningoencephalitis before and after the introduction of the giant African snail. Methods: a study was conducted of a total 19 cerebrospinal fluid and serum samples taken simultaneously from each of the patients diagnosed with eosinophilic meningoencephalitis and kept at the sample collection of the Central Cerebrospinal Fluid Laboratory (LABCEL). Radial immunodiffusion was the laboratory method used for determination of the proteins albumin and IgG. Results: of the patients studied, 14 percent were adults before the appearance of this mollusc, in contrast with the present moment, when 50 percent are patients aged over 18 years. The percentage of mean intrathecal synthesis of IgG was higher in the current sufferers, though not significantly. Conclusions: there are differences before and after the appearance of the giant African snail, given the intrathecal synthesis response of patients associated to the mollusc, which denotes greater aggressiveness by the parasite. The higher mean age of sufferers confirms that these are the ones who most often handle and disperse the mollusc(AU)

El riesgo de parasitismo por Angiostrongylus cantonensis: una problemática reemergente en Cuba / The risk of parasitism by Angiostrongylus cantonensis: a re-emerging problem in Cuba / O risco de parasitismo devido a Angiostrongylus cantonensis: um problema reemergente em Cuba

RESUMEN Introducción: El riesgo de parasitismo por Angiostrongylus cantonensis transmitido por el caracol gigante africano es una problemática reemergente en Cuba. Objetivo: Sistematizar contenidos esenciales relacionados con el caracol gigante africano y el parásito Angiostrongylus cantonensis, el riesgo de meningitis eosinofílica y su manejo. Método: En el Hospital General Docente "Dr. Agostinho Neto", entre septiembre y diciembre de 2019 se hizo una revisión narrativa sobre el tema a través de una búsqueda en bases de datos electrónicas (Biblioteca Virtual en Salud): LILACS, PubMed, SciELO, RedALyC, Scopus, Cochrane. Resultados: La información se estructuró en caracterización del caracol gigante africano y el riesgo de infección por Angiostrongylus cantonensis en Cuba, patogénesis y epidemiologia de la angiostrongylosis, y diagnóstico y tratamiento de la meningitis eosinofílica por este nemátodo. Se identificaron controversias sobre los criterios diagnósticos, el protocolo de actuación para su prevención y rehabilitación, y la carencia de un instrumento para estratificar el riesgo de esta infección. Conclusiones: Se sistematizaron contenidos esenciales sobre el caracol gigante africano, lo que posibilita la preparación de los profesionales para incrementar la percepción del riesgo de infección por Angiostrongylus cantonensis en la población.

ABSTRACT Introduction: The risk of parasitism by Angiostrongylus cantonensis transmitted by the giant African land snail, Lissachatina fulica is a re-emerging problem in Cuba. Objective: Systematize essential contents related to the giant African land snail and the parasite Angiostrongylus cantonensis, the risk of eosinophilic meningitis and its management. Method: A narrative review was made on the subject in the Teaching General Hospital "Dr. Agostinho Neto", between September and December 2019 through a search in electronic databases (Virtual Health Library): LILACS, PubMed, SciELO, RedALyC, Scopus, Cochrane. Results: The information was structured in the characterization of the African giant land snail and the risk of infection by Angiostrongylus cantonensis in Cuba, pathogenesis and epidemiology of angiostrongylosis, and diagnosis and treatment of eosinophilic meningitis by this nematode. Controversies were identified regarding the diagnostic criteria, the protocol of action for its prevention and rehabilitation, and the lack of an instrument to stratify the risk of this infection. Conclusions: Essential content on the giant African land snail was systematized, enabling the preparation of professionals to increase the perception of the risk of infection by Angiostrongylus cantonensis in the population.

RESUMO Introdução: O risco de parasitismo causado por Angiostrongylus cantonensis transmitido pelo caracol gigante africano é um problema reemergente em Cuba. Objetivo: Sistematizar conteúdos essenciais relacionados ao caracol gigante africano e ao parasita Angiostrongylus cantonensis, ao risco de meningite eosinofílica e seu manejo. Método: No Hospital Geral de Ensino "Dr. Agostinho Neto", entre setembro e dezembro de 2019, foi realizada uma revisão narrativa do assunto através de uma busca em bases de dados eletrônicas (Biblioteca Virtual em Saúde): LILACS, PubMed, SciELO, RedALyC, Scopus, Cochrane. Resultados: As informações foram estruturadas na caracterização do caracol gigante africano e no risco de infecção por Angiostrongylus cantonensis em Cuba, patogênese e epidemiologia da angiostrongilose, diagnóstico e tratamento da meningite eosinofílica por esse nematóide. Foram identificadas controvérsias sobre os critérios diagnósticos, o protocolo de ação para sua prevenção e reabilitação e a falta de um instrumento para estratificar o risco dessa infecção. Conclusões: Foram sistematizados conteúdos essenciais sobre o caracol gigante africano, o que possibilita a preparação de profissionais para aumentar a percepção do risco de infecção por Angiostrongylus cantonensis na população.

Evidencias clínico-epidemiológicas vinculadas a Lissachatina fulica en un adulto con meningoencefalitis eosinofílica causada por Angiostrongylus cantonensis / Clinical-epidemiological evidence related to Lissachatina fulica in an adult with eosinophilic meningoencephalitis caused by Angiostrongylus cantonensis

Introducción: El helminto Angiostrongylus cantonensis es un parásito habitual en los pulmones de la rata y puede ocasionar meningoencefalitis eosinofílica en el hombre cuando se pone en contacto con las larvas por ingestión accidental. En Cuba es endémico y el riesgo de contraerlo aumenta con la entrada del caracol gigante africano en el país. Objetivo: Describir las acciones desarrolladas por las autoridades sanitarias locales para el control del caracol gigante africano entre 2016 y 2018 y la aparición de un paciente en 2018 con meningoencefalitis eosinofílica causada por Angiostrongylus cantonensis vinculado epidemiológicamente con la presencia de este caracol. Métodos: Se realizó un estudio cualitativo a partir de un grupo focal con el que se trabajó ante la aparición del caracol gigante africano en un área de salud del municipio San Miguel del Padrón. Se realiza una encuesta semi-estructurada. Resultados: Se trazaron las estrategias para la erradicación de la especie invasora a partir de un trabajo comunitario. Dos años después, se observa nuevamente el caracol gigante africano y un paciente con meningoencefalitis eosinofílica epidemiológicamente asociado a Angiostrongylus cantonensis. Conclusiones: Las acciones realizadas entre 2016 y 2018 resultaron ser insuficientes por el nuevo avistamiento del caracol en el área, con el agravante de encontrar un paciente con meningoencefalitis eosinofílica epidemiológicamente asociado con el molusco(AU)

Introduction: Helmint Angiostrongylus cantonensis is a natural parasite in the lungs of ratas. Ocassionally it can produced an eosinophilic meningoencephalitis in men by larvae accidental ingestion. Methods: A qualitative study was performed from a focal group by a semi-structural survey in a health area from San Miguel del Padrón municipality. Objectives: To describe the actions developed by the local sanitarian authorities for the control of African giant snails between 2016 and 2018 and the appearance of a patient suffering from eosinophilic meningoencephalitis due to Angiostrongylus cantonensis linked to the presence of this snail. Results: It has been established an eradication strategy for the elimination of this invasive species based on a community work. Two years later, it was observed again the giant African snail in the area with a patient suffering from Angiostrongylus cantonensis eosinophilic meningoencephalitis epidemiologically associated. Conclusions: The 2016 actions were not efficient due to the emerging vector and the further finding of a patient linked with the parasite(AU)

Tanshinone IIA attenuates demyelination and promotes remyelination in A. cantonensis-infected BALB/c mice.

BACKGROUND: Angiostrongylus cantonensis infection can cause demyelination in the central nervous system, and there is no effective treatment. METHODS: We used dexamethasone, Tanshinone IIA (TSIIA) and Cryptotanshinone(Two traditional Chinese medicine monomers) in combination with albendazole (AB, a standard anti-helminthic compound) to observe their therapeutic effect on demyelination in A. cantonensis-infected mice. Luxol fast blue staining and electron microscope of myelin sheath, Oligodendrocyte (OL) number and myelin basic protein (MBP) expression in brain was detected in above groups. RESULTS: TSIIA+AB facilitated OL proliferation and significantly increased both myelin sheath thickness and the population of small-diameter axons. In addition, TSIIA treatment inhibited the expression of inflammation-related factors (interleukin [IL]-6, IL-1ß, tumor necrosis factor [TNF]-α, inducible nitric oxide synthase [iNOS]) rather than inhibiting eosinophil infiltration in brain. TSIIA also decreased microglial activation and shifted their phenotype from M1 to M2. CONCLUSIONS: Taken together, these results provide evidence that TSIIA combined with AB may be an effective treatment for demyelination caused by A. cantonensis infection and other demyelinating diseases.

Water transmission potential of Angiostrongylus cantonensis: Larval viability and effectiveness of rainwater catchment sediment filters.

Neuroangiostrongyliasis, caused by Angiostrongylus cantonensis, has been reported in Hawai'i since the 1950's. An increase in cases is being reported primarily from East Hawai'i Island, correlated with the introduction of the semi-slug Parmarion martensi. Households in areas lacking infrastructure for water must use rainwater catchment as their primary domestic water supply, for which there is no federal, state, or county regulation. Despite evidence that slugs and snails can contaminate water and cause infection, regulatory bodies have not addressed this potential transmission route. This study evaluates: 1) the emergence of live, infective-stage A. cantonensis larvae from drowned, non-native, pestiforous gastropods; 2) larvae location in an undisturbed water column; 3) longevity of free-living larvae in water; and 4) effectiveness of rainwater catchment filters in blocking infective-stage larvae. Larvae were shed from minced and whole gastropods drowned in either municipal water or rainwater with ~94% of larvae recovered from the bottom of the water column 72-96 hours post drowning. Infective-stage larvae were active for 21 days in municipal water. Histological sectioning of P. martensi showed proximity of nematode larvae to the body wall of the gastropod, consistent with the potential for shedding of larvae in slime. Gastropod tissue squashes showed effectivity as a quick screening method. Live, infective-stage larvae were able to traverse rainwater catchment polypropylene sediment filters of 20 µm, 10 µm, 5 µm, and 1 µm filtration ratings, but not a 5 µm carbon block filter. These results demonstrate that live, infective-stage A. cantonensis larvae emerge from drowned snails and slugs, survive for extended periods of time in water, and may be able to enter a catchment user's household water supply. This study illustrates the need to better investigate and understand the potential role of contaminated water as a transmission route for neuroangiostrongyliasis.

Infección por Angiostrongylus cantonensis (Nematoda: Metastrongylidae) en Rattus rattus (Rodentia: Muridae) en una localidad de La Habana, Cuba / Infection by Angiostrongylus cantonensis (Nematoda: Metastrongylidae) in Rattus rattus (Rodentia: Muridae) in a locality of Havana, Cuba

Introducción: La meningoencefalitis eosinofílica producida por Angiostrongylus cantonensis (Chen, 1935), es una zoonosis emergente presente en Cuba. En el país existen escasos estudios de prevalencia de infección en los hospederos definitivos, que puedan determinar sitios de riesgo para el establecimiento de medidas efectivas en la prevención de la enfermedad. Objetivo: Determinar la prevalencia de infección por A. cantonensis en una población natural de R. rattus, su relación con la estructura poblacional y la época del año en una finca que pertenece al Programa de la Agricultura Urbana y Suburbana de Cuba. Material y Métodos: Se realizó un estudio descriptivo en el municipio La Lisa, La Habana. Se hicieron capturas de roedores mensuales durante los períodos lluvioso y poco lluvioso. Se examinaron las arterias pulmonares de los roedores capturados en búsqueda de parásitos adultos. El índice parasitológico de prevalencia fue calculado y comparadas las frecuencias entre los períodos del año y la edad ecológica. Resultados: Se capturaron 63 roedores identificados como Rattus rattus, de ellos 46,03 por ciento estuvo infectado con A. cantonensis, con un incremento de la infección en los individuos adultos (59,46 por ciento) y en el período lluvioso (55,88 por ciento). Conclusiones: Se demuestra por primera vez en Cuba la infección de A. cantonensis en R. rattus. La presencia de individuos adultos, la alta abundancia de roedores y la temporada lluviosa son los factores que incidieron en una mayor infección de A. cantonensis, y que aumentan la probabilidad de transmisión a los hospederos intermediarios y, por ende, el riesgo de transmisión al humano(AU)

Introduction: Eosinophilic meningitis caused by Angiostrongylus cantonensis (Chen, 1935) is an emergent zoonosis present in Cuba. In the country, few studies about the prevalence of infection in definitive hosts, which can determine risk sites for the establishment of effective measures to prevent the disease, have not been carried out so far. Objective: To determine the prevalence of infection with Angiostrongylus cantonensis in a natural population of Rattus rattus, its relationship with the population structure, and the season of the year in an urban farm field belonging to the Programme of Urban and Suburban Agriculture of Cuba. Material and Methods: A descriptive study was conducted in La Lisa Municipality, Havana, Cuba. The rats were collected monthly during the rainy and dry seasons. The pulmonary arteries of the captured rodents were examined for adult worms. The prevalence of parasitological indices was calculated. The frequencies of infection were compared between the seasons of the year and ecological ages. Results: A total of 63 rodents identified as Rattus rattus were captured. Of the total of individuals captured, 46.03 percent were infected with A. cantonensis, with an increase of infection in adult individuals (59.46 percent) and during the rainy season (55.88 percent). Conclusions: The infection of A. cantonensis in its definitive host R. rattus, and its circulation in the study area in the analyzed period was demonstrated for the first time in Cuba. The presence of adult rats, the high quantity of rodents and the rainy season were the factors that contributed to a greater infection of A. cantonensis, which increase the probability of transmission to intermediate hosts, and consequently, the risk of transmission to humans(AU)

Increased 14-3-3ß and γ protein isoform expressions in parasitic eosinophilic meningitis caused by Angiostrongylus cantonensis infection in mice.

The 14-3-3 proteins are cerebrospinal fluid (CSF) markers of neuronal damage during infectious meningitis and Creutzfeldt-Jakob disease. Little is known about dynamic changes in the individual isoforms in response to parasitic eosinophilic meningitis. The purposes of this study were to determine the 14-3-3 protein isoform patterns, examine the kinetics and correlate the severity of blood brain barrier (BBB) damage with the expressions of these markers in mice with eosinophilic meningitis. Mice were orally infected with 50 A. cantonensis L3 via an oro-gastric tube and sacrificed every week for 3 consecutive weeks after infection. The Evans blue method and BBB junctional protein expressions were used to measure changes in the BBB. Hematoxylin and eosin staining was used to analyze pathological changes in the mice brains following 1-3 weeks of infection with A. cantonensis. The levels of 14-3-3 protein isoforms in serum/CSF and brain homogenates were analyzed by Western blot, and immunohistochemistry (IHC) was used to explore the different isoform distributions of 14-3-3 proteins and changes in BBB junctional proteins in the mice brain meninges. Dexamethasone was injected intraperitoneally from the seventh day post infection (dpi) until the end of the study (21 dpi) to study the changes in BBB junctional proteins. The amounts of Evans blue, tight junction and 14-3-3 protein isoforms in the different groups of mice were compared using the nonparametric Kruskal-Wallis test. There were significant increases in 14-3-3 protein isoforms ß and γ in the CSF in the second and third weeks after infection compared to the controls and first week of infection, which were correlated with the severity of BBB damage in brain histology, and Evans blue extravasation. Using IHC to assess the distribution of 14-3-3 protein isoforms and changes in BBB junctional proteins in the mice brain meninges, the expressions of isoforms ß, γ, ε, and θ and junctional proteins occludin and claudin-5 in the brain meninges increased over a 3-week period after infection compared to the controls and 1 week after infection. The administration of dexamethasone decreased the expressions of BBB junctional proteins occludin and claudin-5 in the mice brain meninges. Our findings support that 14-3-3 proteins ß and γ can potentially be used as a CSF marker of neuronal damage in parasitic eosinophilic meningitis caused by A. cantonensis.

Severe CNS angiostrongyliasis in a young marine: a case report and literature review.

Angiostrongylus cantonensis is the most common cause of eosinophilic meningitis worldwide. Infection typically occurs through ingestion of undercooked molluscs or vegetables contaminated by infective larvae. Endemic regions were previously limited to southeast Asia and the Pacific basin; however, this parasite is seeing an alarming increase in global distribution with reported cases in more than 30 countries, including several states in the USA. Although infection typically results in meningitis, a broad spectrum of CNS involvement and severity is emerging as diagnostic methods (such as real-time PCR) continue to improve diagnosis. In this Grand Round, we report a case of a 20-year-old active duty US marine serving in Okinawa, Japan, afflicted with severe CNS angiostrongyliasis marked by radiculomyelitis with quadriparesis, hyperaesthesia, and urinary retention. We present this case to highlight that no clear guidelines exist for the treatment of severe CNS angiostrongyliasis and provide our consensus recommendation that treatment algorithms include use of dual corticosteroids plus anthelmintics when radicular symptoms are present. In this Grand Round we review the clinical features, epidemiology, advances to diagnostic techniques, and available data on current treatment options for CNS angiostrongyliasis. This diagnosis should be highly considered in the differential diagnosis of a patient presenting with meningeal symptoms, paraesthesia or hyperaesthesia, and CSF eosinophilia so that treatment can be started early, which is particularly important in children, because of their increased risk of severe disease and mortality. We recommend combined therapy with albendazole and prednisolone, with consideration for increased steroid dosing in severe cases.