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. soleiformis. 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

Variability in the fecal egg count and the parasitic burden of hair sheep after grazing in nematode infected paddocks

This study aimed to evaluate the variability in the fecal egg count (FEC) and the parasitic burden of naive hair sheep after grazing in nematode infected paddocks. The research was carried out in Tabasco, Mexico, during two periods (August and December). In each period 32 lambs were grazed for one month on African star grass (Cynodon plectostachyus) contaminated with gastrointestinal parasitic nematodes. FEC, packed cell volume (PCV) and body weight (BW) were recorded. Gastrointestinal worms were recovered at necropsy. Data were analyzed with the MIXED procedure of SAS using a model of repeated measurements over time. A higher number of Haemonchus contortus worms was found in December (2814±838) than in August (1166±305). The opposite occurred with Cooperia curticei (2167±393 and 3638±441, respectively). The FEC and correlation coefficient in respect to the worm burden were higher in December (6516 ± 1599, r=0.83, respectively) than in August (4364±771, r=0.44, respectively). A high variability in resistance-susceptibility to gastrointestinal nematodes (GIN) occurred in Katahdin × Pelibuey lambs after grazing.

Efficacy of an enzyme-linked immunosorbent assay as a diagnostic tool for schistosomiasis mansoni in individuals with low worm burden

IgM-ELISA is an immunoenzymatic method useful for detection of IgM antibodies against a fraction of Schistosoma mansoni adult worm antigen (AWA) that is soluble in trichloroacetic acid (AWA-TCA). This method was applied to three groups of individuals with different clinical and epidemiological characteristics, and the results compared with those obtained by other diagnostic methods: immunofluorescence test for detection of IgM antibodies (IgM-IFT) or IgG antibodies (IgG-IFT), ELISA for detection of IgG antibodies (IgG-ELISA), and two parasitological methods, Kato-Katz and miracidium hatching. The IgM-ELISA presented a sensitivity of 98%, when the parasitologic fecal examination was defined as reference diagnostic method, and a specificity of 98 and 97.3%, respectively for the group of clinically healthy individuals and other helminth carriers. A comparative analysis between the results of IgM-ELISA and those obtained by other serologic tests showed a good degree of agreement, with Kappa indices ranging from 0.95 to 0.98. The diagnostic efficacy of 97.8%, as determined with schistosomiasis patients with low parasitic burden, suggests the excellent performance of the IgM-ELISA and its usefulness for the diagnosis of schistosomiasis when applied in low endemic areas.

Longitudinal clinical and serological survey of abdominal angiostrongyliasis in Guaporé, southern Brazil, from 1995 to 1999

Abdominal angiostrongyliasis is a zoonotic infection caused by Angiostrongylus costaricensis, a nematode with an intra-vascular location in the mesentery. Our objective was to address several aspects of the natural history of this parasitosis, in a longitudinal clinical and seroepidemiological study. A total of 179 individuals living in a rural area with active transmission in southern Brazil were followed for five years (1995-1999) resulting in yearly prevalence of 28.2%, 4.2%, 10%, 20.2% and 2.8% and incidences of 0%, 5.9%, 8% and 1.5%, respectively. Both men and woman were affected with higher frequencies at age 30-49 years. In 32 individuals serum samples were collected at all time points and IgG antibody reactivity detected by ELISA was variable and usually persisting not longer than one year. Some individual antibody patterns were suggestive of re-infection. There was no association with occurrence of abdominal pain or of other enteroparasites and there was no individual with a confirmed (histopathologic) diagnosis. Mollusks were found with infective third-stage larvae in some houses with an overall prevalence of 16% and a low parasitic burden. In conclusion, abdominal angiostrongyliasis in southern Brazil may be a frequent infection with low morbidity and a gradually decreasing serological reactivity.

Pre- and post-treatment immunodiagnostic evaluation in human schistosomiasis mansoni

Schistosomiasis control seems to be different in countries were low parasitic burden and asymptomatic clinical patients are the features of majority of cases. Immunological methods must substitute the traditional coprologic techniques used for some decades in the Control Program. Circumoval Precipitin Test (COPT), intradermal test and ELISA with soluble egg antigen (SEA) are evaluated for using as tools for seroepidemiologic studies. COPT and ELISA were performed after treatment to known their utility when impact of chemotherapy must be assessed. One hundred sixty five persons were followed up 3, 6, 9 and 12 months after treatment. The mean sensitivity of CPT studied by age groups was 95.6% which is very important considering that 88.4% of the studied population excreted less than 100 egg/gr of feces, while sensitivity of intradermal test was 58.2%. Children showed the highest ractivity to COPT. When treatment is effective, COPT reactivity progressively disminish until become negative one year later. In the non cure group, the COPT reactivity disminished but never below 20%. ELISA-SEA did not modify one year after treatment. Effort should be made to isolate fractions of eggs Schistosoma mansoni whose antibodies disappear after treatment.

An analysis of the impact of the Schistosomiasis Control Programme in Brazil

The impact of the Schistosomiasis Control Programme (PCE) in Brazil was analyzed, covering the period 1976 to 2003, using the following indicators: percentage of Schistosoma mansoni carriers detected among the population examined in the coproscopic surveys (PPS): mortality rate for schistosomiasis, per 100,000 inhabitants (TME): hospitalization rate for schistosomiasis, per 100,000 inhabitants (TIE): average age of deaths caused by schistosomiasis (IMOE). There was a 38.5% reduction in the PPS after the introduction of the PCE, attributed to the treatment of carriers. Even in hyper-endemic municipalities, such as Conde and Cuitegí, in the state of Paraíba, the PPS fell more than 50% after the first year of treatment. The parasitic burden of the carriers also decreased in the two municipalities. The TME was reduced by 63.4% and the TIE by 77.3%. The mortality rate was highest among the 50-and-above age group. The country×s IMOE rose 32.3%. The IMOE was seen to be much lower in the state of Minas Gerais, where the PCE was only initiated in 1983, with very limited coverage.

Low susceptibility of Achatina fulica from Brazil to infection with Angiostrongylus costaricensis and A. cantonensis

Introduction of Achatina fulica in Brazil has led to serious concerns about its role as vector for metaIylid worms: AngioIylus costaricensis and A. cantonensis. Experimental infection with both parasites was performed to evaluate the potential risk for their transmission by the giant African snail. Groups of 5 animals, both wild and bred at captivity were exposed at different inocula: 1, 5, and 10 ´ 10³ L1 of A. costaricensis and A. cantonensis. In all groups, few snails got infected and parasitic burden was low. Two different ways of infection were tested: ingestion produced higher numbers of L3 than the inoculation through an artificial hole in the shell. We also report the parasitological examination of 6 batches of wild A. fulica from Florianópolis, state of Santa Catarina, Brazil: only 1 out of 244 animals were infected with metaIylid larvae. Taken together these data indicate that the giant African snail occurring in Southern Brazil is not a permissive host for both AngioIylus species and does not represent a significant risk for transmission of these parasites.

Le Baylisascaris procyonis dans le sud du Québec : prévalence chez le raton laveur et étude de contamination des latrines

Le raton laveur (Procyon lotor) est porteur de nombreux parasites dont certains sont des agents de zoonose. Il nous est apparu opportun d’étudier la faune parasitaire intestinale du raton laveur québécois, particulièrement l’espèce Baylisascaris procyonis. L’étude a donc porté sur l’examen de matières fécales (n = 301) et de contenus intestinaux (n = 203) de 351 ratons laveurs provenant de deux programmes gouvernementaux sur la rage du raton laveur et d’excréments (n = 409) provenant de latrines dans des parcs nationaux du Québec. Des oeufs de B. procyonis étaient excrétés par 23,3 % des ratons laveurs, alors que 29,1 % étaient porteurs de vers. La charge parasitaire moyenne des vers B. procyonis de tout stade variait de 8,5 chez les adultes à 27,1 chez les juvéniles. Chez ces derniers, la charge parasitaire était 11,4 fois supérieure à celle des ratons adultes (p < 0,0001) dont 23,7 fois plus de vers immatures (p < 0,0001) et 9,1 fois plus de vers matures (p = 0,01). Le nombre d’oeufs était 24,7 fois plus élevé chez les ratons juvéniles (p = 0,02) et 50,0 fois moins élevé chez la femelle allaitante. La probabilité du raton d’excréter des oeufs l’automne était de 11,1 (IC95 %: 1,21-101,60) par rapport au printemps et de 21,1 (IC95 %: 2,91-153,18) par rapport à l’été (p < 0,01). La sensibilité (Se) et la spécificité (Sp) de la coproscopie comparée à la nécropsie (norme étalon) ont été calculées en se basant sur l’observation des vers matures (Se : 81,8 %; Sp : 97,7 %) et des vers de tout stade (Se : 53,9 %; Sp : 97,0 %). De plus, la recherche des parasites intestinaux nous a permis de découvrir des ookystes de Giardia (6/159), espèce parasitaire nouvellement rapportée chez le raton. En 2007, nous avons trouvé, par échantillonnage unique, des oeufs de B. procyonis dans 15,8 % des latrines (n = 165) identifiées dans des parcs nationaux du sud du Québec et en 2008, par échantillonnage multiple, dans 89,7 % des latrines (n = 26) actives situées dans des secteurs accessibles à l’homme dans trois parcs nationaux de la Montérégie. Le potentiel zoonotique de B. procyonis est un problème de santé publique qui pourrait devenir sérieux étant donné le pourcentage élevé de ratons laveurs qui excrètent le parasite et qui contaminent des secteurs accessibles par les humains.

Infecções naturais em cervídeos (Mammalia: Cervidae) procedentes dos Estados do Mato Grosso do Sul e São Paulo, por nematódeos Trichostrongyloidea Cram, 1927

No período compreendido entre 1985 e 1996 foram necropsiados, para pesquisa de helmintos, 42 cervídeos, sendo sete Mazama americana, 16 M. gouazoubira, 13 Ozotoceros bezoarticus e seis Blastocerus dichotomus. Desses animais, foram colhidos 14.426 nematódeos Trichostrongyloidea, sendo 13.281 (92,06%) parasitos de abomaso e 1.145 (7,94%), de intestino delgado. Nesses órgãos, foram identificadas seis espécies de nematódeos: Haemonchus contortus, H. similis, Trichostrongylus axei, T. colubriformis, Cooperia punctata e C. pectinata. Todos os animais apresentaram infecções helmínticas por uma ou mais espécies, ocorrendo grande variação na intensidade de infecção (1 a 4.345 nematódeos). Ainda com relação à intensidade de infecção, os dados expressavam valores menores que 100 parasitos em 25 (59,52%) animais. Os valores mais altos de intensidade média das infecções foram observados em M. gouazoubira (596,37 helmintos) e em O. bezoarticus (331), e os menores, em M. americana (17,57) e B. dichotomus (75,5). Os dados mais expressivos de intensidade de infecção, abundância e prevalência foram observados para Haemonchus (larvas de 4º estágio), H. contortus, H. similis e T. axei. O gênero Haemonchus foi constatado em 35 animais, com prevalência de 83,33%; apresentou carga parasitária de 11.616 exemplares, representando 80,52% dos nematódeos verificados, sendo a maioria (8.903) constituída por formas imaturas. Por outro lado, H. similis foi a espécie predominante nas infecções e, portanto, a que apresentou maiores valores de abundância. Verificou-se o gênero Trichostrongylus em 24 (57,14%) animais, com carga parasitária de 2.444 exemplares, sendo 1.665 espécimes de T. axei, que representou 11,54% da carga parasitária obtida. As seis espécies de vermes identificadas nos cervídeos são comuns aos ruminantes domésticos nos Estados de São Paulo e Mato Grosso do Sul e dessa maneira não se observou nenhuma espécie de Trichostrongyloidea exclusiva dos cervídeos.

Immunotherapy against murine experimental visceral leishmaniasis with the FML-vaccine

The fucose mannose ligand (Leishmania donovani FML)-saponin vaccine has earlier shown its immunoprophylactic potential against visceral leishmaniasis in the CB hamster (87.7% of parasite load reduction), Balb/c (84.4%) and Swiss albino mouse (85-93%) models. In this investigation its specific immunotherapeutic efficacy against L. donovani infection in Balb/c mice was studied. The effects of vaccine treatment on the Immoral response, delayed type of hypersensitivity to promastigote lysate (DTH), cytokine levels in sera and reduction of the liver parasitic load of L. donovani infected mice, were examined. The types and subtypes of anti-FML antibodies increased significantly in the vaccinees over the saline and saponin controls. As expected for a saponin vaccine, the highest ratios were found in relation to IgG1, IgG2a and IgG2b (4.4, 5 and 2.5, respectively). The DTH response and the in vitro ganglion cell proliferative response against FML antigen were also significantly higher than controls (P < 0.005). Concomitantly, an impressive and specific decrease of liver parasitic burden was detected only in vaccine-treated animals (94.7%). Our results indicate that the therapeutic FML-vaccine has a potent effect on modulation of the murine infection leading to the reduction of parasitic load and signs of disease, being a new potential tool in the therapy and control of visceral leishmaniasis. (C) 2003 Elsevier Ltd. All rights reserved.

Endectocide activity of a new long-action formulation containing 2.25% ivermectin+1.25% abamectin in cattle

The present work aimed to evaluate the endectocide activity of a new injectable long-action formulation, containing ivermectin (IVM) and abamectin (ABA). In each one of the four experiments performed, the following groups were formed: group I: 2.25% IVM (450 mu g/kg) + 1.25% ABA (250 mu g/kg), group II: 3.15% IVM (630 mu g/kg) and group III: control. Eighteen bovine naturally infected by gastrointestinal nematoda were selected for anthelmintic evaluation and necropsied on posttreatment day (PTD) 14 to estimate the total parasitic burden. For the Rhipicephalus (Boophilus) microplus field trial, 30 bovine were selected by means of counts of semi-engorged R. (B.) microplus and the therapeutic and residual efficacy evaluated by tick counts on PTDs 1, 3, 7, 14, 21, 28, 35, 42, 49, 56, 63, 70, 77, 84 and 91. In the stall test, 15 calves were artificially infested with 5000 R. (B.) microplus (Mozzo strain) larvae three times a week and daily collections of all the engorged female ticks detached from each calf were performed until the PTD 80. Forty bovine naturally infected with Dermatobia hominis larvae were selected and the number of larvae was counted by visual and tactile inspection on PTDs 3, 7, 14, 28, 35, 49, 63, 77, 91 and 105. In this trial, a formulation containing 1% doramectin (200 mu g/kg) was also used. IVM + ABA formulation and 3.15% IVM eliminated four of the eight species of nematode identified. The anthelmintic efficacy of the avermectins association against Haemonchus placei, Cooperia spatulata and C. punctata was 89.64%, 98.84% and 97.69%, while 3.15% IVM achieved 30.98%, 84.79% and 75.56%, respectively. The two formulations evaluated showed reduced acaricide action on the PTD I and 3, reaching high efficacy percentages from PTD 14 onward. The IVM + ABA showed efficacy above 95% in the period between PTDs 21 and 49. In the stall test, it observed no difference (P > 0.05) between the two formulations regarding the R. (B.) microplus counts during the entire evaluation period. IVM + ABA reduced the number of ticks from the PTD 1 to 77 (P < 0.05) and 3.15% IVM reduced (P < 0.05) the tick number from PTD 4 up to PTD 80. The three endectocides showed no difference (P > 0.05) regarding the number of D. hominis larvae and prevented this parasite reestablishment until PTD 105. These results indicate that the IVM + ABA association showed higher anthelmintic activity and similar efficacy against arthropods to the formulation containing 3.15% IVM. (C) 2008 Elsevier B.V. All rights reserved.

Anthelmintic efficacy of an oral formulation of Aurixazol against gastrointestinal nematodes of naturally and experimentally infected sheep

As a result of the need to develop new active principles for the control of endoparasites in ruminants, the present in vivo study evaluated a formulation containing 24% Aurixazol (48 mg/kg), a parasiticide molecule based on disophenolate of levamisole. Two experiments were conducted: one evaluating the anthelmintic efficacy of 24% Aurixazol (48 mg/kg) against gastrointestinal nematodes in naturally infected sheep, compared to an association of ivermectin (0.2 mg/kg) + albendazole (5.0 mg/kg) + levamisole (7.5 mg/kg) (IAL), and a second one which evaluated the persistent efficacy of the same formulation against immature stages (L4) and adults of Haemonchus contortus in experimentally infected animals. In experiment I, against H. contortus, the formulation of Aurixazol and the IAL association reached efficacies (arithmetic means) of 99.32% and 96.11%, respectively. For Trichostrongylus colubriformis, the efficacy values were 88.92% and 98.08% for Aurixazol and the IAL association, respectively. Both formulations were totally effective against Oesophagostomum columbianum (100%). The results of the statistical analysis demonstrated that the mean parasitic burden of treated animals was significantly different (P ≤ 0.05) compared to the average number of helminths diagnosed in animals from the control group for H. contortus, T. colubriformis and O. columbianum. Comparing only the treated groups, it was possible to verify that the average number of H. contortus recovered from animals treated with Aurixazol was different (P ≤ 0.05) when compared to the mean amount recovered from sheep treated with the IAL association. When evaluating the prevention of H. contortus infection in experiment II, Aurixazol did not present preventive efficacy. Up until 21 days after treatment the groups treated with Aurixazol contained less adults and L4 of H. contortus (P ≤ 0.05) when compared to the non-medicated control group. However, future studies will be necessary to assess the effectiveness of Aurixazol against nematode strains resistant to levamisole and disophenol, but the efficacy results described in this study allow to state that Aurixazol can, associated with other measures, become an important tool in the control of sheep nematodes. © 2013.

Nematode resistance to ivermectin (630 and 700 mu g/kg) in cattle from the Southeast and South of Brazil

Two high doses of ivermectin (630 mu g/kg and 700 mu g/kg) that are sold commercially in Brazil were evaluated in dose-and-slaughter trials with 144 naturally nematode-infected cattle from eight regions within the states of Minas Gerias, Sao Paulo and Rio Grande do Sul in Brazil. Treatment groups were based on fecal egg counts 1, 2, and 3 days before treatment; all animals studied had a minimum egg count of at least 500 eggs per gram of feces (EPG). Post-mortem analyses were conducted on day 14. The highest levels of resistance to ivermectin were found for Haemonchus placei, Cooperia punctata and Oesophagostomum radiatum; all populations of H. placei were resistant to the 630 mu g/kg dose, and 67% were resistant to 700 mu g/kg; 86% of C. punctata were resistant to the 630 mu g/kg dose, and 33% were resistant to 700 mu g/kg. A combined analysis revealed that 57% of O. radiatum were resistant to the lower dose of ivermectin. H. placei, C. punctata and O. radiatum, in order, were the nematode populations with the highest indices of resistance, whereas Trichostrongylus axei was the most susceptible to 630 and 700 mu g/kg dosages of ivermectin. The results of helminthic resistance to ivermectin for different populations of H. placei and C. punctata described in the present study support previous literature data, in which a small decrease in the average parasitic burden of C. punctata and a consequent increase of H. placei were observed in cattle from the Southeast, South and Center-West regions of Brazil. (C) 2014 Elsevier Ireland Ltd. All rights reserved.

MyD88 and TLR9 dependent immune responses mediate resistance to Leishmania guyanensis infections, irrespective of Leishmania RNA virus burden.

Infections with Leishmania parasites of the Leishmania Viannia subgenus give rise to both localized cutaneous (CL), and metastatic leishmaniasis. Metastasizing disease forms including disseminated (DCL) and mutocutaneous (MCL) leishmaniasis result from parasitic dissemination and lesion formation at sites distal to infection and have increased inflammatory responses. The presence of Leishmania RNA virus (LRV) in L. guyanensis parasites contributes to the exacerbation of disease and impacts inflammatory responses via activation of TLR3 by the viral dsRNA. In this study we investigated other innate immune response adaptor protein modulators and demonstrated that both MyD88 and TLR9 played a crucial role in the development of Th1-dependent healing responses against L. guyanensis parasites regardless of their LRV status. The absence of MyD88- or TLR9-dependent signaling pathways resulted in increased Th2 associated cytokines (IL-4 and IL-13), which was correlated with low transcript levels of IL-12p40. The reliance of IL-12 was further confirmed in IL12AB-/- mice, which were completely susceptible to infection. Protection to L. guyanensis infection driven by MyD88- and TLR9-dependent immune responses arises independently to those induced due to high LRV burden within the parasites.

Experimental infection of Leishmania chagasi in immunosuppressed balb/c mice: cellular immune response and parasite burden

The immune response to leishmaniasis can result in a polarization of a subpopulation of T lymphocytes, which leads to a different cell phenotype and results in immune protection or exacerbation of the disease. Leishmanias persist in the body both in asymptomatic infections and after treatment, representing risks in terms of immunosuppression. The objective of this study was to evaluate the effects of infection and immunosuppression by dexamethasone associated with pentoxifylline on animal weight, spleen weight, the parasitic load in the spleen and liver, as well as the production of IFN-gamma and IL-10 in spleen cell culture of Balb/c mice infected with Leishmania chagasi. The infection did not alter animal weight gain, but spleen weight and size increased. The immunosuppression, induced by dexamethasone associated with pentoxifylline, affected animal weight gain and weight and size of the spleen (in infected and not infected animals). The immunosuppression did not significantly alter the course of the parasite burden in the spleen and liver. Dexamethasone and pentoxifylline affected the studied cytokine production, but not influenced on Th1/Th2 response in infected animals.