Costaricensis Infection Research Articles

Diagnostic criteria and case definitions for abdominal angiostrongyliasis: a systematic review from the Brazilian experience.

Although rare, Angiostrongylus costaricensis infection may be a more prevalent etiology of inflammatory bowel disease than ulcerative colitis and Chron's disease in endemic areas in Central and South America. The present study reviewed the occurrence of A. costaricensis in Brazil, its clinical presentation and pathology; and proposed diagnostic criteria and case definitions for abdominal angiostrongyliasis (AA). Southern and southeastern Brazilian regions are the main endemic areas, and AA affects both genders and all age groups. A review of all 23 published reports of 51 Brazilian patients highlighted the following features that were subsequently classified as minor diagnostic criteria: abdominal pain, palpable mass in the right lower abdominal quadrant, history of exposure, ileocecal tumor, and intestinal perforation with wall thickening. Proposed major criteria include right lower quadrant abdominal pain, blood eosinophilia, positive serology (antibody detection), intense eosinophilic infiltration that involves all strata of the intestinal wall, eosinophilic granulomatous reaction, and eosinophilic vasculitis. In addition to the definitions of suspected and possible cases according to increasing strength of evidence of this infection, demonstration of worms/eggs/larvae in tissues or Angiostrongylus DNA in tissues or serum are required for a confirmed diagnosis. The application of the proposed criteria and definitions may improve patient management, epidemiologic surveillance, and identification of new endemic areas.

Relationship between histopathological lesions and oxidative stress in mice infected with Angiostrongylus costaricensis.

This study describes changes in oxidative stress (OS) parameters in mice experimentally infected with Angiostrongylus costaricensis, which causes abdominal angiostrongyliasis. For this, 28 Swiss mice were used, divided into two groups (G1 and G2), with 14 animals each. Of these, eight were infected with ten infective larvae each, by gavage, and six were used as a control group. Mice from G1 and G2 were euthanized at 14 days and 24 days post-infection, respectively. Tissue samples were used for histopathological analysis and blood (serum) samples were taken to assess the levels of proteins, non-protein thiols (NPTs) and nitric oxide (NO), from centrifugation and subsequent collection of aliquots of the supernatant. Among OS parameters, infected mice in both groups had higher NO levels than the control group, due to the presence of: eosinophil infiltrate in the liver and intestine; pancreatitis; and intestinal granuloma. However, the infected mice of both groups showed a reduction in the levels of NPTs, in relation to the control group, due to the presence of: eosinophilic infiltrate in the liver and intestine; and intestinal granuloma. Our results suggest that A. costaricensis infection has important effects on the intestine, liver and pancreas, and the analyses were performed from the tissue of these organs. The mechanisms for these changes are related to the decrease in the body's main antioxidant defences, as demonstrated by the reduction of NPTs, thus contributing to the development of more severe tissue damage. Thus, the objective of the present study was to evaluate the relationship between histopathological lesions and markers for OS.

Abdominal angiostrongyliasis can be diagnosed with a immunochromatographic rapid test with recombinant galactin from Angiostrongylus cantonensis.

Angiostrongylus costaricensis is the causative agent of abdominal angiostrongyliasis, a zoonotic infection that may produce severe eosinophilic enterocolitis or hepatitis in humans. Parasites are usually not released in stools and serology has an important role in diagnosis. Since cross-reactivity is demonstrated between A. costaricensis and another metastrongylid worm, A. cantonensis, we tested heterologous recombinant galectin as a probe in an immunochromatographic rapid diagnostic test (ICT-RDT) for detection of anti-A. costaricensis antibodies. Almost all (11/12) positive control sera from A. costaricensis infected patients were positive at ICT RDT. These are preliminary indications that r-galectin ICT-RDT is useful for diagnosing A. costaricensis infection.

The white-nosed coati (Nasua narica) is a naturally susceptible definitive host for the zoonotic nematode Angiostrongylus costaricensis in Costa Rica

Angiostrongylus costaricensis (Strongylida, Angiostrongylidae) is a roundworm of rodents, which may cause a severe or fatal zoonosis in several countries of the Americas. A single report indicated that the white-nosed coati (Nasua narica), acts as a potential free-ranging wildlife reservoir. Here we investigated the prevalence and features of A. costaricensis infection in two procyonid species, the white-nosed coati and the raccoon (Procyon lotor) from Costa Rica to better understand their possible role in the epidemiology of this zoonotic infection. Eighteen of 32 (56.2%) white-nosed coatis collected between July 2010 and March 2016 were infected with A. costaricensis but none of 97 raccoons from the same localities were diagnosed with this infection. First-stage larvae of A. costaricensis were obtained from feces of 17 fresh white-nosed coati carcasses by Baermann technique. Parasite identity was confirmed by morphology, histology and molecular characterization of target genes. These data demonstrate that the white-nosed coati is a naturally susceptible definitive host for A. costaricensis in Costa Rica contrary to findings in the raccoon.

Elimination of Angiostrongylus costaricensis larvae in feces from experimentally infected Swiss mice: circadian rhythm and correlation with survival

Angiostrongylus costaricensis is a nematode which harbors mesentery arteries of rodents. In these animals, a circadian rhythm of elimination of first-stage larvae (L1) and a relation between the amount of L1 in feces and survival are unknown. We assessed fecal elimination of A. costaricensis L1 from experimentally infected Swiss mice and tried to correlate L1 elimination with survival. Thirteen Swiss mice were infected by gavage with ten A. costaricensis L3 larvae obtained from Phyllocaulis slugs. Feces were weighed at 7 A.M: . and 7 P.M: . starting from the 24th day post-infection until animal death. Feces sediment was examined in microscope for L1 counting. The mice were dead after a period ranging 19-61 days post-infection. Compared to diurnal samples, both feces' weight (2.3 ± 0.7 vs. 1.8 ± 0.5 g; P < 0.0001) and L1 total count [median 1,950 vs. 1,250; P = 0.015] were higher in feces eliminated at night. No difference was observed between diurnal and nocturnal elimination when counting L1 by gram of feces (725 vs. 650 L1/g; P = 0.821). A significant correlation was observed between survival and total number of L1 in feces (r = 0.84; P = 0.0007). This study suggests that mice experimentally infected with A. costaricensis eliminate more L1 at night due to higher fecal volume at this period. The correlation between number of L1 in feces and survival suggests a phenomenon of tolerance to A. costaricensis infection in mice with longer survival.

Alteration in the endogenous intestinal flora of swiss webster mice by experimental Angiostrongylus costaricensis infection

The association between worm infections and bacterial diseases has only recently been emphasized. This study examined the effect of experimental Angiostrongylus costaricensis infection on endogenous intestinal flora of Swiss Webster mice. Eight mice aging six weeks were selected for this experiment. Four were infected with A. costaricensis and the other four were used as controls. Twenty eight days after the worm infection, all mice in both groups were sacrificed and samples of the contents of the ileum and colon were obtained and cultured for aerobic and anaerobic bacteria. In the mice infected with A. costaricensis there was a significant increase in the number of bacteria of the endogenous intestinal flora, accompanied by a decrease in the number of Peptostreptococcus spp. This alteration in the intestinal flora of mice infected by the nematode may help to understand some bacterial infections described in humans.

Long-term observations on mouse strains experimentally infected with Angiostrongylus costaricensis.

BALB/c and C57BL/6 mice were experimentally infected with Angiostrongylus costaricensis and the parasitic parameters and antibody response during the acute and chronic phases of infection were analyzed. Following administration of six third-stage larvae (L3), there was no significant difference in the mean worm recovery or mean larval output. Coinciding with the maturation of worms in infected animals and with the egg output in mesenteric arteries, a strong increase in the humoral immune response was observed in both mouse strains. This response was characterized by a hypergammaglobulinemia, with a predominance of IgA and IgG1 during the acute phase of infection, and IgG1 and total IgE during the patent and post-patent periods. Significantly higher levels of IgM, IgG and IgG1 were found in BALB/c mice compared with C57BL/6 mice. On the other hand, a significantly higher concentration of IgA was detected at 6 and 7 weeks post-infection in C57BL/6 mice compared with BALB/c mice. Specific IgE could not be detected in any of the mouse strains. Our results suggest that immunoglobulins, mainly IgG1, contribute to the outcome of a primary A. costaricensis infection with respect to the period of patency and to mortality during the chronic phase.

Infection of BALB/c mice with Angiostrongylus costaricensis decreases pulmonary inflammatory response to ovalbumin.

SUMMARY The prevalence of asthma in developing countries is lower than in developed countries. Viral, bacterial and parasitic infections may be associated with this discrepancy. The relationship between parasitic infection and asthma prevalence is not clear. Previous controversial data have demonstrated that parasitic infection may either predispose or protect against the development of asthma. The aim of this study is to determine whether infection with Angiostrongylus costaricensis (A. costaricensis) decreases inflammatory lung response to ovalbumin (OVA) in mice. Seven BALB/c mice were infected with A. costaricensis by orogastric gavage (10 larvae/mouse) on day (D) 0. The mice were immunized against OVA by intraperitoneal injection on D 5 and D 12 and received an intranasal OVA challenge (40 micro L) on D 15 and D 17. On D 19 bronchoalveolar lavage (BAL) was performed. Six BALB/c mice (control group) were immunized with OVA using the same protocol, but were not infected with A. costaricensis. Interleukin (IL)-1beta and IL-6 levels were measured in the BAL fluid by using commercial ELISA assays. Total cell counts and differential cell counts were performed in the BAL fluid samples. The group infected with A. costaricensis had lower total cell count in the BAL fluid when compared with the control group (0.11 x 10(6)cells/mL and 0.3 x 10(6)cells/mL, respectively; P = 0.013). BAL fluid IL-1beta levels in the infected group were significantly lower than in the control group (P = 0.008). IL-6 levels in BAL fluid were not different between the groups studied. We conclude that Angiostrongylus costaricensis infection in mice decreases pulmonary inflammatory response to OVA.

Angiostrongylus costaricensis infection in C57BL/6 mice: MHC-II deficiency results in increased larval elimination but unaltered mortality.

During experimental Angiostrongylus costaricensis infections in several inbred mouse strains, genetic factors as well as different cytokine secretion patterns have recently been shown to play a role in the outcome of infection in terms of morbidity and mortality, e.g. BALB/c mice show a high and C57BL/6 mice a low mortality during the acute phase of infection. In this study, C57BL/6 MHC-II knockout mice infected with A. costaricensis did not show increased mortality during the acute phase of infection when compared with wild-type mice. Furthermore, MHC-II knockout mice showed a strongly diminished parasite-specific humoral and cellular immune response, which can be explained by the nearly complete lack of CD4+ T cells in the periphery. This defect in MHC-II genes, the lack of CD4+ T cells, and the resulting cellular and humoral unresponsiveness resulted in a three times higher output of first-stage larvae in feces compared with wild-type animals. The results indicate that during experimental A. costaricensis infection a parasite-specific immune response, directed via MHC-II molecules and CD4+ T cells, is not essential for the survival of C57BL/6 mice during the acute phase of infection, whereas the elimination of first-stage larvae seems to be regulated by a MHC-II- and CD4+ T-cell-dependent mechanism.

Kinetics of eosinophil and IgE-mast cell changes following infection with Angiostrongylus costaricensis in Wistar rats.

Human abdominal angiostrongyliasis is a severe eosinophilic disease caused by Angiostrongylus costaricensis. Previous studies have demonstrated that wild rodents are critically involved as definitive hosts to this nematode in nature. In this study, we have evaluated the susceptibility of Wistar rats (Rattus norvegicus) to A. costaricensis infection. Kinetics of parasitological and pathological changes, including the number of adult worms recovered from mesenteric arteries, and of IgE, mast cell and eosinophil levels in several compartments have been assessed. The oral inoculation of third-stage larvae (L3) into adult Wistar rats led to a marked accumulation of worms in the branches of the mesenteric arteries 25 and 50 days post-inoculation. Intense bone marrow eosinophilia ranging from 7 to 50 days was accompanied by marked accumulation of eosinophils in the blood, peritoneal and bronchoalveolar spaces. Eosinophilic periarteritis, oedema and granuloma in the intestinal and lung tissues were also histologically evident. Total serum IgE and specific anti-parasite IgE peaked at 25 days post-infection, as measured by ELISA and by the passive cutaneous anaphylaxis test, respectively. At that time point, there was a drastic reduction in the number of intact mast cells in the peritoneal effluent. These findings indicate that Wistar rats are permissive to A. costaricensis infection. IgE-mast cell activation and massive tissue eosinophil infiltration are marked features in the process and are likely to play a crucial role in the immune-response evoked by this parasite.

Intestinal granuloma formation in normal and SCID BALB/c mice infected with Angiostrongylus costaricensis.

We studied the histomorphology of granuloma formation and cytokine production in Angiostrongylus costaricensis-infected immunocompetent and immunodeficient BALB/c mice. Histological examination of the infected intestine showed well developed granulomas in BALB/c mice. In contrast, in SCID mice, clusters of eggs and larvae and a progression of infiltration of inflammatory cells were also observed, but little fibroblastic activity was seen. Analysis of plasma of the infected mice demonstrated a dramatic contrast in the cytokine profile between normal and SCID mice. Normal mice infected with A. costaricensis showed highly elevated plasma IFN-gamma levels at week 3 of infection, but plasma IL-4 remained close to the background levels obtained from non-infected mice. In contrast, the amount of those cytokines in the plasma of SCID animals was little affected by this parasitic infection. These results suggest that Th1-mediated immunity is required for granuloma formation in response to A. costaricensis infection in mice.

Interleukin-5 transgenic mice show augmented resistance to Angiostrongylus costaricensis infection.

To determine the possible role of eosinophils in Angiostrongylus costaricensis infection, both interleukin-5 (IL-5) transgenic (Tg) and non-transgenic (non-Tg) C3H/HeN mice were infected with A. costaricensis third-stage larvae. IL-5 Tg mice demonstrated greater resistance than non-Tg mice to A. costaricensis, as shown by lower adult worm recovery, smaller adults, fewer eggs in the intestinal wall and fewer larvae passed in the feces. Both mice showed similar antigen-specific IgA and IgGI antibody responses, although IgA was more prominent than IgG1. Egg deposition and inflammatory responses in the intestinal walls were milder in IL-5 Tg mice than in non-Tg mice. The eggs with developed larvae, deposited in the intestinal walls of IL-5 Tg mice, were surrounded by numerous degranulating eosinophils and sometimes with Splendore-Hoeppli deposits. The data suggest that eosinophils are involved in the resistance of the mouse during primary infection with A. costaricensis.

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-γ) 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.

Cyclooxygenase-2-derived prostaglandin E2 and lipoxin A4 accelerate resolution of allergic edema in Angiostrongylus costaricensis-infected rats: relationship with concurrent eosinophilia.

In noninfected rats, challenge with allergen following local IgE sensitization induced a pleurisy marked by intense protein exudation that plateaued from 30 min to 4 h after challenge, reducing thereafter. Infection of rats with Angiostrongylus costaricensis induced a 5-fold increase in blood eosinophil numbers by 25 days postinfection, whereas the numbers of eosinophils in the pleural cavity ranged from normal to a weak increase. In infected rats, identically sensitized, challenge with Ag induced a much shorter duration of pleural edema with complete resolution by 4 h, but no change in the early edema response. In parallel, infection increased the number of eosinophils recovered from the pleural cavity at 4 h, but not at 30 min, following allergen challenge. Pretreatment with IL-5 (100 IU/kg, i.v.) also increased eosinophil numbers in blood and, after allergen challenge, shortened the duration of the pleural edema and increased pleural eosinophil numbers. There were increases in the levels of both PGE2 and lipoxin A4 (LXA4) in pleural exudate. Selective cyclooxygenase (COX)-2 inhibitors, NS-398, meloxicam, and SC-236, did not alter pleural eosinophilia, but reversed the curtailment of the edema in either infected or IL-5-pretreated rats. Pretreatment of noninfected animals with the PGE analogue, misoprostol, or two stable LXA4 analogues did not alter the magnitude of pleural exudation response, but clearly shortened its duration. These results indicate that the early resolution of allergic pleural edema observed during A. costaricensis infection coincided with a selective local eosinophilia and seemed to be mediated by COX-2-derived PGE2 and LXA4.

Experimental Angiostrongylus costaricensis infection in mice: immunoglobulin isotype responses and parasite-specific antigen recognition after primary low-dose infection.

Immunoglobulin isotype responses and parasite-specific antigen recognition were investigated in experimental Angiostrongylus costaricensis infection in two different mouse strains. Even in a low-dose infection with third-stage larvae (L3), BALB/c mice showed high mortality until 28 days postinfection (p.i.) in association with a low patency rate in surviving animals. On the other hand, low mortality and a high rate of patent infection was observed in C57BL/10 mice. Parasite-specific IgM, total IgG, and IgG subclasses against crude adult-worm antigen (AcAg) rose in both groups of mice from day 14 onward, with IgG and IgG1 being significantly elevated in BALB/c mice at 21 and 28 days p.i., respectively. For total IgE, significantly elevated concentrations were detected at 14 days p.i. in BALB/c mice as compared with C57BL/10 mice. A. costaricensis-specific antigen recognition by total IgG, IgG1, or IgG2a was similar in both mouse strains, intensifying from 3 to 4 weeks p.i., with recognition of immunodominant AcAg ranging between 80 and 210 kDa. This study provides evidence that in BALB/c and C57BL/10 mice, immunoglobulins, with the possible exception of IgE and IgG1, do not decisively contribute to the outcome of a primary A. costaricensis infection with respect to immunopathogenesis or parasite permissiveness.

Immune defense and eosinophilia in congenitally IgE-deficient SJA/9 mice infected withAngiostrongylus costaricensis

The roles of IgE in protective immunity and eosinophilia in Angiostrongylus costaricensis infection were examined by comparing IgE-deficient SJA/9 mice and IgE-producing SJL/J mice. In primary infection, mean total IgE levels increased to a maximum of 390 ng/ml, which was more than 10 times greater than the 29 ng/ml measured preinfection in SJL/J mice but less than the 10 ng/ml found in SJA/9 mice throughout the experiment. Immune defense as determined by recovery of adult worms and eosinophilia were similar in SJL/J and SJA/9 mice. Protective immunity was induced by infection with A. costaricensis followed by treatment with levamisole for 4-6 days postinfection. After the challenge infection, the numbers of adult worms and eosinophils in SJA/9 mice were not significantly different from those in SJL/J mice. Anti-A. costaricensis IgE antibody was not detected in either strain of mice during the experiment. These results indicate that A. costaricensis infection induced the production of IgE not specific for parasite antigens in IgE-producing mice. Potentiated nonspecific IgE played no significant role in immune defense and eosinophilia.

Strain-dependent differences in susceptibility of mice to experimental Angiostrongylus costaricensis infection

Experimental Angiostrongylus costaricensis infection was carried out in inbred strains of mice (C57BL/6 BALB/c, DBA/2 and C3H/He). All strains became infected with this parasite. Marked differences in mortality and in worm burden were found among inbred strains of mice tested. A significant reduction was shown in worm length from mice compared to that from cotton rats.