Evaluating host-parasite co-adaptation relationships involving Angiostrongylus costaricensis.

Angiostrongylus costaricensis is a parasite that infects rodents, including the wild cotton rat Sigmodon hispidus and pygmy rice rats Oligoryzomys spp., among others. However, urban Rattus norvegicus and Mus musculus have not been identified as important hosts of A. costaricensis. In this study, Swiss mice (SW), Wistar R. norvegicus (RN), wild Oligoryzomys nigripes (ON) and a local strain of M. musculus (RGS) were experimentally infected with A. costaricensis. Survival, elimination of L1 (total sum per group, A0), and the number of adult worms recovered divided by the dose of each L3 inoculum (yield ratio, YR) were examined for each group after a 40-day post-infection period. The survival rates, A0 and YR values were: 27%, 207,589 and 0.42 for the SW group; 81%, 8691 and 0.01 for the RN group; and 63.6%, 26,560 and 0.16 for the RGS group, respectively, in each case. The survival rate for the ON group was 100% and the A0 value was 847,050. A YR was not calculated for the ON group since the ON group was maintained up to 565 days post-infection (pi) to examine long-term mortality. At 500 days pi (16 months), 50% of the ON group had died, while one animal (10%) survived 595 days pi (20 months). Taken together, these data indicate that A. costaricensis has undergone a greater degree of adaptation to the wild rodent, O. nigripes, than to R. norvegicus or a local M. musculus strain. In addition, titre curve (A0) modelling of adaptation status proved to be useful in evaluating A. costaricensis-rodent interactions.

Invasive slug Meghimatium pictum (Stoliczka, 1873) infected by Angiostrongylus costaricensis Morera & Céspedes, 1971, and the possible risk of human infection associated with grape consumption.

Many molluscs may be infected with angiostrongylid larvae. Following the histopathological diagnosis of abdominal angiostrongyliasis in a grape farmer from southern Brazil, molluscs in the area were investigated. During a nocturnal search, 245 specimens of slugs were collected and identified as the invasive Chinese slug Meghimatium pictum. Angiostrongylus costaricensis worms were recovered from mice that were experimentally infected with larvae obtained from 11 (4.5%) of the molluscs. This study presents the first report of M. pictum being identified as an intermediate host for A. costaricensis. Most of the slugs were collected from grape plants, which suggests that transmission may be associated with grape consumption.

Effect of different helminth extracts on the development of asthma in mice: The influence of early-life exposure and the role of IL-10 response.

It is not currently clear whether different parasites have distinct effects on the airway inflammatory response in asthma and whether exposure in early life to helminths have a stronger impact in a potential inhibitory effect on asthma. The aim of this study is to evaluate the effect of exposure to different helminth extracts on the development of allergic pulmonary response in mice, including early-life exposure. Different helminth extracts (Angiostrongylus costaricensis, Angiostrongylus cantonensis and Ascaris lumbricoides) were studied in female adult BALB/c and C57BL/6 IL-10-deficient mice in a protocol of murine asthma, injected intraperitoneally in different periods of exposure (early, pre-sensitization and post-sensitization). Cell counts in bronchoalveolar lavage (BAL), eosinophil peroxidase (EPO) from lung tissue, cytokine levels from BAL/spleen cell cultures, and lung histology were analyzed. Airway cellular influx induced by OVA was significantly inhibited by extracts of A. cantonensis and A. lumbricoides. Extracts of A. lumbricoides and A. costaricensis led to a significant reduction of IL-5 in BAL (p < 0.001). Only the exposure to A. lumbricoides led to an increased production of IL-10 in the lungs (p < 0.001). In IL-10-deficient mice exposed to A. costaricensis pre-sensitization, eosinophil counts and IL-5 levels in BAL and EPO in lung tissue were significantly reduced. In the early exposure to A. cantonensis, lung inflammation was clearly inhibited. In conclusion, different helminth extracts inhibit allergic lung inflammation in mice. IL-10 may not play a central role in some helminth-host interactions. Early exposure to helminth extracts could be a potential strategy to explore primary prevention in asthma.

Abdominal angiostrongyliasis in rodent experimental infection: evidence for systemic circulation of first stage larvae.

Eggs of Angiostrongylus costaricensis embrionate and hatch in the course of their migration in the intestinal wall, and first stage larvae (L1) are released in feces. Aiming to investigate the possible systemic circulation of L1, we inoculated mice and, four weeks later, examined their peritoneal cavities and several organs for the presence of L1. A total of 65 larvae were found in extra-intestinal organs (kidney, heart, spleen, liver, lungs). No larvae were found in blood or in the peritoneal cavity. 320 and 578 L1 were found in intestinal wall and intestinal contents, respectively. The experiment was repeated and it confirmed that the metastrongylid larvae found in several organs were larval stages of A. costaricensis. Ten Oligoryzomis sp. rodents, a natural host for A. costaricensis, were also infected and in three animals L1 could be recovered from several organs as well as from bronchoalveolar lavage (BAL) in one of them. These data indicate that systemic circulation and bronchial elimination of L1 may represent an alternative route for release of L1 of A. costaricensis into the environment for transmission to the intermediate host.

Inactivation of infective larvae of Angiostrongylus costaricensis with short time incubations in 1.5% bleach solution, vinegar or saturated cooking salt solution.

Angiostrongylus costaricensis is a nematode parasite of wild rodents in the Americas. Man may become infected accidentally and, sometimes, suffers a very severe abdominal disease. Ingestion of raw vegetables has been proven to be a risk factor for the acquisition of A. costaricensis and, therefore, prophylaxis should include food disinfection. The larvicidal effect of wine vinegar, saturated cooking salt (SS) and a 1.5% bleach solution (BW) were compared with a 1-h-incubation period, at room temperature. Larval viability was tested through inoculation in Swiss mice. Only one out of 560 larvae treated with BW (97.3% of the animals were uninfected) was recovered as an adult worm, while 90/336 and 29/512 larvae treated, respectively, with SS and WV were recovered as adult worms. This larvicidal effect of BW was seen also in incubation times as short as 15 min. In conclusion, the 1.5% bleach solution may be helpful for prophylaxis of human abdominal angiostrongyliasis through disinfection of raw vegetables and unpeeled fruits.

Cellular immune responses and cytokine production in BALB/c and C57BL/6 mice during the acute phase of Angiostrongylus costaricensis infection.

In our experimental study we were able to show that the contrasting outcome of Angiostrongylus costaricensis infection in C57BL/6 and BALB/c mice, in respect of morbidity and mortality, can be explained by divergent cellular immune responses and a different cytokine pattern in each strain. In BALB/c mice (i.e. those with high mortality), the initial high proliferation of ConA or LPS stimulated spleen cells dropped to very low levels after 2 weeks post-infection (p.i.), whereas in C57BL/6 mice (i.e. those with low mortality), only a minor reduction in lymphoproliferative responses after mitogenic stimulation was observed. The specific proliferation of spleen cells after stimulation with A. costaricensis adult worm antigen remained low in BALB/c mice throughout the experiment, but showed an augmented proliferation in C57BL/6 mice, especially from 2 weeks p.i. onwards. The mitogen-induced production of Th2-type cytokines (IL-4, IL-5, IL-6, IL-10) in spleen cell cultures remained low in BALB/c mice until 4 weeks p.i., but production of Th1-type cytokines (IL-2, IFN-gamma) was highly elevated at 14 and 28 days p.i. In C57BL/6 mice, an upregulated and balanced production of both Th1- and Th2-type cytokines was measured during the course of infection. In summary, a polarization of the immune response towards cellular hyporesponsiveness and a predominantly Th1 cytokine profile was observed in A. costaricensis infected BALB/c mice, which may contribute to pathogenesis and increased morbidity.

Isolation of Angiostrongylus costaricensis first-stage larvae from rodent feces on a Percoll gradient.

Several density gradients were tested for the isolation of parasitic nematode, Angiostrongylus costaricensis, first-stage larvae from rodent feces. With a 45/72% Percoll gradient, 83-99% (89.56+/-6.57%) of the larvae were recovered in a clean preparation.

Clinical and epidemiological aspects of abdominal angiostrongyliasis in southern Brazil.

Most of the cases of abdominal angiostrongyliasis in Brazil were reported from the southern States of São Paulo, Paraná, Santa Catarina and Rio Grande do Sul (RS). A study in 27 cases from RS revealed a distinct local epidemiology. Peasants were usually affected, either adults or children, from the mountainous areas in the north of the State. There was a seasonal increase in the number of cases, from late spring to autumn, that does not coincide with the rainy season. Besides the most common clinical features of abdominal pain, fever and eosinophilia in the leukogram, painful relapsing episodes were detected in some patients. The abdominal pain could be either localized or diffuse during the rapid evolution to a surgical abdominal condition, with a lethality of 7.4%. The use of a serological test and the greater awareness of physicians working in endemic areas is expected to improve the recognition of uncomplicated and benign courses of the disease. This study confirms the known clinical manifestations of abdominal angiostrongyliasis and demonstrates the diversity of its epidemiology.

Evidences against a significant role of Mus musculus as natural host for Angiostrongylus costaricensis.

Wild rodents have been described as the most important hosts for Angiostrongylus costaricensis in Central America and southern Brazil. Sinantropic rodents apparently do not play a significant role as natural hosts. A search for natural infection failed to document worms in 14 mice captured in the house of a patient with diagnosis of abdominal angiostrongylosis and experimental infection of a "wild" Mus musculus strain and groups of albino swiss mice were carried out. Mortality was not significantly different and varied from 42% to 80% for Swiss mice and from 26% to 80% for "wild" mice. The high mortality of a "wild" M. musculus infected with A. costaricensis was very similar to what is observed with most laboratory mice strains. These data may be taken as indications that M. musculus is not a well adapted host for A. costaricensis, although susceptibility was apparently higher with "wild" populations of M. musculus as compared to Swiss strain.

The effect of temperature on mobility of Angiostrongylus costaricensis third stage larvae.

Third stage larvae (L3) from Angiostrongylus costaricensis were incubated in water at room temperature and at 5 degrees C and their mobility was assessed daily for 17 days. Viability was associated with the mobility and position of the L3, and it was confirmed by inoculation per os in albino mice. The number of actively moving L3 sharply decreased within 3 to 4 days, but there were some infective L3 at end of observation. A mathematical model estimated 80 days as the time required to reduce the probability of infective larvae to zero. This data does not support the proposition of refrigerating vegetables and raw food as an isolated procedure for prophylaxis of human abdominal angiostrongylosis infection.

[Identification of wild rodents as hosts of Angiostrongylus costaricencis in southern Brazil]. / Identificação de roedores silvestres como hospedeiros do Angiostrongylus costaricensis no sul do Brasil.

Increasing number of human cases of abdominal angiostrongyliasis has been diagnosed in the south of Brazil. The main definitive host of Angiostrongylus costaricensis in Central America is the cotton rat (Sigmodon hispidus) that does not occur in South America, except in the north of Colombia, Peru and Venezuela. Rodents were captured in the endemic area in Rio Grande do Sul (RS) and definitive hosts were identified for the first time in Brazil: Oryzomys nigripes and Oryzomys ratticeps. O. nigripes is a small wild rodent and it appears to be the main definitive host of A. costaricensis in the highlands of RS, Brazil's southernmost State.

Angiostrongylus costaricensis and the intermediate hosts: observations on elimination of L3 in the mucus and inoculation of L1 through the tegument of mollucs.

Human accidental infection with Angiostrongylus costaricensis may result in abdominal disease of varied severity. Slugs from the Veronicellidae family are the main intermediate hosts for this parasitic nematode of rodents. Phyllocaulis variegatus, Phyllocaulis soleiformis and Phyllocaulis boraceiensis were experimentally infected to describe the kinetics of L3 elimination in the mucus secretions of those veronicelid species. A maximum of 2 L3/g/day was found in the mucus, while the number of L3 isolated from the fibromuscular tissues varied from 14 to 448. Productive infection was established by inoculations in the hyponotum or in the body cavity, through the tegument. Intra-cavity injection is a less complex procedure and permits a better control of inocula. A preliminary trial to titrate the infective dosis for P. variegatus indicated that inocula should range between 1000 and 5000 L1. The data also confirmed the importance of P. variegatus as an intermediate host of A. costaricensis.

[Abdominal angiostrongyliasis: its prevention by the destruction of infecting larvae in food treated with salt, vinegar or sodium hypochlorite]. / Angiostrongilose abdominal: profilaxia pela destruição das larvas infectantes em alimentos tratados com sal, vinagre ou hipoclorito de sódio.

There is a high prevalence of accidental human infection with Angiostrongylus costaricensis in some areas in southern Brazil and sometimes it presents as severe intestinal disease. Prophylaxis is important since there is no medical treatment for the disease. The ingestion of fruits and vegetables contaminated with the mucous secretion of infected molluscs (the intermediate hosts) is one of the proposed modes of transmission. Third stage larvae were incubated at 5 degrees C for 12 hours, in solutions of saturated sodium chloride, vinegar and sodium hypochlorite 1.5%. The larvae had their viability tested through inoculation into albino mice. The percentage of larvae that established infection were 0% in the group treated with sodium hypochloride, 1.8% with NaCl and 2.4% with vinegar. In conclusion, all substances tested reduced the population of viable larvae and may be useful in food decontamination, as a prophylactic measure for abdominal angiostrongylosis.

[The using of hollow bricks as a shelter for adaptation of wild rodents' colonies in captivity]. / Tijolos vazados como abrigo para adaptação de roedores silvestres em cativeiro.

Difficulties for adaptation of wild rodents' colonies in the laboratory may hamper experimental studies. We describe the use of a hollow brick inside cages, to provide shelter and reduce stress of the animals. It facilitates the cleaning of cages, since the animals stay inside the brick while they are transferred to a new cage. This method was applied in a trial for adaptation of Oryzomys nigripes in captivity. Despite transportation for more than 1500 km, mortality was zero in the group of 62 rodents and the colony expanded immediately. A hollow brick is a cheap and widespread available material for improving adaptation of wild rodents to captivity.

Prospective study of strongyloidosis in patients with hematologic malignancies.

Immunocompromised individuals infected with Strongyloides stercoralis may develop severe hyperinfection or disseminated disease with high mortality. Patients with hematological malignancies are at risk because of immunodepression produced either by the disease or its treatment. A prospective study was undertaken at the Hospital de Clínicas de Porto Alegre, from July 1994 to July 1995. Seventy-two (HIV negative), had 3 stool samples collected at different days and had not received recent anthelmintic therapy. Larvae, isolated in a modified Baermann method, were found in 6 patients, with a resultant prevalence of 8.3%. No complicated strongyloidosis was documented. The positive result for S. stercoralis larvae was significantly associated (p < 0.001) with eosinophilia. Knowledge of prevalence figures and incidence of severe disease is important to adequate guidelines for empirical treatment besides the rigorous search for strongyloidosis in patients with hematological malignancies.

Report on the occurrence of Angiostrongylus costaricensis in southern Brazil, in a new intermediate host from the genus Sarasinula (Veronicellidae, Gastropoda).

Veronicellid slugs are the main intermediate hosts for Angiostrongylus costaricencis. In a rural locality in Nova Itaberaba (SC, southern Brazil) Sarasinula linguaeformis was identified as a crop pest. The parasitological examination revealed A. costaricencis infection in 43 out ot 50 slugs. The prevalence of 86% and the individual parasitic burdens are the highest sofar reported in Brazil and S. linguaeformis is the first species from the genus Sarasinula to be identified as intermediate host for A. costaricencis in southern Brazil.

Histopathological criteria for the diagnosis of abdominal angiostrongyliasis.

An increasing number of cases of abdominal angiostrongyliasis have recently been diagnosed in southern Brazil. A comparative study of 19 confirmed cases and 15 suspected cases was undertaken to review the anatomo-pathology of the disease and to establish histopathological criteria for its diagnosis. The results were similar in both groups, except for the identification of the worm in the confirmed cases. Macroscopic features comprised two types: a predominant thickening of the intestinal wall (pseudo-neoplastic pattern) and a congestive necrotic lesion (ischemic-congestive pattern). Microscopically, three fundamental histopathological findings were detected: (1) a massive infiltration of eosinophils in all layers of the intestinal wall, (2) a granulomatous reaction and (3) eosinophilic vasculitis affecting arteries, veins, lymphatics and capillaries. We conclude that observation of this histopathological triplet excludes other causes of "eosinophilic gastroenteritis" and establishes the diagnosis of probable abdominal angiostrongyliasis.

Abdominal angiostrongylosis in southern Brazil--prevalence and parasitic burden in mollusc intermediate hosts from eighteen endemic foci.

Angiostrongylus costaricensis is a parasitic nematode of rodents and molluscs are the intermediate hosts. Nocturnal collection of molluscs and search for infective third stage larvae of A. costaricensis was carried out in 18 endemic foci identified by the notification of a confirmed diagnosis in human biopsies or surgical specimens. Molluscs were digested in acidic solution and isolation of larvae eventually present was done in a Baermann funnel. Larvae identified by the presence of a delicate groove in the tail were counted to assess the individual parasitic burden. Four species were found infected, with ranges of prevalence in parenthesis: Phyllocaulis variegatus (7% to 33.3%); Bradybaena similaris (11.7% to 24.1%); Belocaulus angustipes (8.3%) and Phyllocaulis soleiformis (3.3% to 14.2%). Parasitic burden varied from 1 to 75 with P. variegatus, 1 to 98 with B. similaris. 1 to 13 with B. angustipes and 1 larvae in each of two specimens of P. solciformis. P. variegatus was present in all sites and was found infected with the highest prevalence figures and the highest individual parasitic burdens. These data stress the importance of veronicellid slugs as intermediate hosts for A. costaricensis in the endemic areas in Rio Grande do Sul, Brazil.

Study on the elimination of Angiostrongylus costaricensis first stage larvae in the experimental infection of Swiss mice.

Abdominal angiostrongylosis is a nematode infection wild rodents. Human infection may result in severe abdominal disease and has been reported from several countries in the Americas. The domestic mouse, Mus musculus, has not been found with natural infection and, like other urban rodents, should not be considered a natural host for Angiostrongylus costaricensis. Quantification of parasitic forms released for transmission may better express the coevolutionary status in parasite-host relationship. With this objective, five groups of experimentally infected Swiss mice were followed for up to 155 days post-infection (PI) days and the quantification of first stage larvae (L1) output revealed: an irregular elimination of L1 and a huge variation in the patency period (1 to 114 days) and in the number of L1 eliminated daily by individual animals (1 to 6340 L1/g). Overall mortality was 72% (range: 28% to 100%) at seven weeks PI. In conclusion, abdominal angiostrongylosis in M. musculus presents high mortality and a very variable and irregular elimination of L1 in feces.