Using freshwater snail Biomphalaria glabrata (Say, 1818) as a biological model for ecotoxicology studies: a systematic review.

Over time, a growing increase in human pollutants in the aquatic environment has been observed. The global presence of residues in water bodies reinforces the need to develop improved methods to detect them and evaluate their ecotoxicological effects in aquatic environments. Thus, this study aimed to present the main assays using Biomphalaria glabrata as a biological model for ecotoxicological studies. We performed a systematic literature review with data published up to June 2022 on the Web of Science, SCOPUS, Science Direct, PubMed, and SciELO databases. Thirty studies were selected for this review after screening. Biomphalaria glabrata has been studied as an ecotoxicological model for different substances through toxicity, embryotoxicity, cytotoxicity, genotoxicity, and bioaccumulation assays. Studies evaluating the impact of B. glabrata exposure to several substances have reported effects on their offspring, as well as toxicity and behavioral and reproductive effects. This review presents various assays using B. glabrata as a biological model for ecotoxicological studies. The use of a representative species of ecosystems from tropical regions is a necessary tool for tropical environmental monitoring. It was observed that the freshwater snail B. glabrata was effective for the evaluation of the ecotoxicity of several types of chemical substances, but further studies are needed to standardize the model.

Parasitism in Rattus rattus and sympatric Achatina fulica by Angiostrongylus cantonensis in an urban park in southeast Brazil.

In this study, rodents (Rattus rattus) and mollusks (Achatina fulica) were captured in a small forest located in a large metropolitan city in Brazil, and they were examined to investigate possible parasitism by Angiostrongylus cantonensis. The parasites were recovered as helminths from the pulmonary arteries of the synanthropic rodents and as third-stage larvae (with Metastrongylidae family characteristics) from the mollusks. To confirm the species, these larvae were used to experimentally infect Rattus norvegicus for the posterior recovery of adult helminths. To identify the adult helminths, morphological, morphometric, molecular, and phylogenetic techniques were employed. Furthermore, we also characterized the histological lesions associated with parasitism in naturally infected definitive hosts. Our results demonstrated the occurrence of a natural life cycle of A. cantonensis (with the presence of adult helminths) in definitive hosts, Rattus rattus, and third-stage larvae in an intermediate host, A. fulica. In free-living rodents, lesions of granulomatous pneumonia in the lungs and meningitis in the brain were also found. These results warn of the risk of accidental transmission of A. cantonensis to human residents around the park because of the extensive interaction among the fauna of the park, domestic animals, and the surrounding human population.

Identification of a phenoloxidase- and melanin-dependent defence mechanism in Achatina fulica infected with Angiostrongylus vasorum.

BACKGROUND: Angiostrongylus vasorum has different freshwater aquatic and terrestrial gastropod molluscs as an intermediate host, e.g. Arion spp. The mollusc Achatina fulica is a danger to public health, given the large diversity of nematodes utilizing it as an intermediate host, such as the parasites of the genus Angiostrongylus, of importance in human and veterinary medicine. Achatina fulica has been shown to have an excellent capacity for maintaining outbreaks and natural infections with A. cantonensis in Asia. Within the mollusc, the nematode parasites activate haemocytes and/or haemolymph factors and in some invertebrates, phenoloxidase (PO), that induces the release of toxic elements and eliminates the parasites. Despite the importance of A. fulica in the life-cycle of nematodes, little is known regarding the defence mechanisms involving PO in molluscs infected with nematodes. Here, the presence of PO and nitric oxide (NO) in the haemolymph and haemocytes of A. fulica infected with first-stage (L1) larvae of Angiostrongylus vasorum was evaluated, together with the presence of melanin in the cephalopod mollusc tissue. RESULTS: An increase in PO at one day post infection (dpi), in comparison with the control using the substrates L-tyrosine (F(4,90) = 6.73, P = 0.00006), L-DOPA (F(4,90) = 22.67, P = 0.02) and p-phenylenediamine (PPD) (F(4,90) = 27.58, P = 0.0019), was observed. PO increase coincided with the presence of melanin in the cephalopodal tissue. At 8 dpi, PO activity, compared to L-DOPA (F(4,90) = 22.67, P = 0.00002) and PPD (F(4,90) = 27.58, P = 0.079) decreased, while melanin increased. At 13 dpi, PO decreased with PPD (F(4,90) = 27.58, P = 0.000015) and also the amount of melanin observed in histology. At 30 dpi, PO increased along with the substrates L-DOPA and PPD, while melanin decreased. NO levels increased until 8 dpi, and decreased after 13 dpi. CONCLUSIONS: To our knowledge, this is the first study that illustrates PO activity in a helminth-infected A. fulica and provides the first observation of an L-tyrosine dependent PO activity in molluscs infected with A. vasorum. This work suggests that PO pathway may help to control A. vasorum infection in A. fulica.

Rangelia vitalii in a free-ranging maned wolf (Chrysocyon brachyurus) and co-infections.

An adult free-ranged female maned wolf was rescued from a periurban area subject to anthropogenic disturbances in the Minas Gerais, Brazil. The animal presented poor body condition and anemia. The clinical condition rapidly deteriorated culminating in dead and a necropsy was performed. The main gross lesions were marked anemia and blood content in the intestines accompanied by many types of parasites. The protozoa Rangelia vitalii was identified by histopathological analysis predominantly within the cytoplasm of endothelial cells of capillaries of the small intestine. The lymph nodes, spleen, bone marrow, dermis, lungs and kidney had similar protozoal forms but with mild or moderate intensity. Rangelia vitalii was confirmed by molecular assays. Hepatozoon sp., Leishmania sp., and Entamoeba spp., apparently not related to the clinical signs were also detected. The myriad parasites found in the intestines included nematodes (Ancylostoma caninum, A. braziliensis,, Molineus sp., Pterygodermatites sp., and Trichuris sp.), cestodes (Spirometra sp.) and (acanthocephalans. To our knowledge, R. vitalii was identified in C. brachyurus for the first time. These findings emphasize the fragility of Brazilian ecosystems, especially in disturbed areas, reinforcing the necessity of efforts to preserve these areas and wild carnivores, some of which are threatened with extinction, such as the maned wolf.

Plants of the Cerrado naturally selected by grazing sheep may have potential for inhibiting development of Haemonchus contortus larva.

Plant species naturally selected by sheep grazing in the Cerrado region of Brazil were assessed in vitro for activity against Haemonchus contortus. One year of observations showed the plant families in the region exhibiting greatest richness to be Fabaceae, Rubiaceae, Malpighiaceae, Bignoniaceae, Myrtaceae, and Annonaceae. Nine species commonly selected by grazing sheep showed variation in the selectivity index with respect to the dry and rainy seasons. Coproculture was conducted in five replicates of 11 treatments: ivermectin, distilled water, or dehydrated leaves of nine selected plant species administered at 333.3 mg g(-1) fecal culture. The dried powder of Piptadenia viridiflora and Ximenia americana leaves significantly reduced the number of infective larvae compared to the distilled water control. These species showed efficacy of over 85 % despite low concentrations of proanthocyanidin. High-performance liquid chromatography analyses of extracts of these plants showed major peaks of UV spectra characteristic of flavonoids. Those naturally selected plant species with high antihelminthic efficacy show promise for use in diet as an alternative control of H. contortus in sheep.

Proteolytic activity of the nematophagous fungus Arthrobotrys sinensis on Angiostrongylus vasorum larvae.

BACKGROUND: The predatory nematophagous fungus Arthrobotrys sinensis (SF53) produces three proteases with nematicidal activity when grown on solid media culture. However, the proteolytic profile produced by this fungus, when grown in liquid culture medium remains unknown. FINDINGS: Thus, the objective of this work was to evaluate the production of proteases from nematophagous fungus Arthrobotrys sinensis in liquid medium and its nematicidal activity on first stage larvae of A. vasorum. Proteases were obtained in its crude form, using Whatman no.1 filter paper, followed by centrifugation for 5 min at 10 × g and 4°C. A zymogram was performed with co-polymerized casein in an acrylamide gel as substrate. An in vitro assay to evaluate the nematicidal action of the proteases of A. sinensis (SF53) produced in liquid medium on A. vasorum L1 was conducted. By the analysis of the zymogram, it was observed a single halo at the beginning of digestion of the gel, suggesting that the three proteases of SF53 are produced in an enzymatic complex of large molecular weight. Regarding nematicidal activity, within 24 hours, the proteases produced in liquid medium of A. sinensis (SF53) showed a percentage reduction of 64% on the number of L1 of A. vasorum. CONCLUSION: In the present work, it is suggested that the three proteases of SF53 are produced in an enzymatic complex and was also demonstrated that these enzymes were effective in destroying A. vasorum L1.

In vitro activity of a serine protease from Monacrosporium thaumasium fungus against first-stage larvae of Angiostrongylus vasorum.

A serine protease from the nematophagous fungus Monacrosporium thaumasium (NF34a) was purified, partially characterized and tested in vitro in control of the first larval stage of Angiostrongylus vasorum. NF34a grew in liquid culture medium, producing its crude extract that was purified by ion exchange chromatography. The fractions with high protease activity were collected in a pool, and elution of proteases was monitored by enzymatic assay and protein content. Purification steps were monitored by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Protease activity was determined under different pH and temperature conditions, and the inhibitor effects of metal ions and phenylmethylsulfonyl fluoride (PMSF) were assessed. In an experimental test, the infection process of NF34a on first-stage larvae of A. vasorum was investigated. A purified serine protease (Mt1) was identified, with an approximate molecular mass of 40 kDa and apparent homogeneity in SDS-PAGE, having optimal activity at pH 7.0 to 8.0 and temperature of 60°C. Mg(2+) and Zn(2+) partially inhibited the activity of Mt1 while PMSF inhibited it completely. Mt1 production was observed when NF34a was grown using first-stage larvae of A. vasorum as the only source of carbon and nitrogen. These results show that the enzyme may have a possible role in the infection process of the larvae. In the in vitro test of applicability against A. vasorum L(1), we observed a reduction in the number of larvae of 23.9% (p < 0.05) in the group treated with Mt1 compared with the control group. However, even this low reduction demonstrates that the Mt1 is important in the infection process.

Effect of temperature on activity of third-stage larvae of Angiostrongylus vasorum.

To evaluate the effect of temperature on the activity and mortality of the L3 of Angiostrongylus vasorum, 1,500 L3 were isolated from experimentally infected snails and distributed into five equal groups. Three groups were incubated at 37°C, 27°C, and 5°C. The remaining two groups were incubated at 27°C and 5°C for 10 days, at which time the temperature for the 27°C group was reduced to 5°C and the 5°C group increased to 27°C. Larva activity was observed daily and inactive larvae were removed. At 37°C, larvae survived up to 8 days. At 27°C, larvae were active until day 6. When subjected to a reduction in temperature from 27°C to 5°C beginning on day 10, the number of active larvae increased until day 13. Only on day 17 did the number of active larvae decline to zero. At 5°C, larvae remained active until day 15, surviving to 24 days. When temperature was increased from 5°C to 27°C beginning on day 10, larvae were found active until day 12 and maintained an intermediate level of activity to day 21. Survival of larvae was greater at lower temperatures, while high temperatures were associated with higher mortality.

Wild and domesticated animals as reservoirs of Schistosomiasis mansoni in Brazil.

Natural infection with Schistosoma mansoni in wild vertebrates and domesticated animals in Brazil is described in this review from an epidemiological viewpoint. Some species of wild rodents are small-sized animals, with a short expectation of life, a limited territory, and present high infection rates under natural conditions. A successful maintenance of the parasite's life cycle under artificial conditions can be achieved with Biomphalaria glabrata. On the other hand, despite showing low natural infection rates, cattle are very susceptible to infection under experimental conditions (using calves of Holstein lineage, cross-bred with the Gir lineage). Due to their large size (just one calf may harbor a number of worms higher than a whole colony of aquatic rodents) and their longevity, cattle are a potential reservoir for the maintenance and dissemination of the disease. There is thus a need of new studies to gain a better understanding about the actual role of these animals in the epidemiology of S. mansoni.

Identification of stage-specific proteins of Angiostrongylus vasorum (Baillet, 1866) Kamensky.

Sera of ten dogs infected with Angiostrongylus vasorum were analyzed by immunoblotting proteins of first stage larvae (L1) and adult parasites. The molecular weights (m.w.) of the principal L1 proteins identified by IgG, IgM, IgA, and IgE were 18-118 kDa and those of the adult parasite were 28-209 kDa. The L1 proteins had not been recognized by IgG, IgM, and IgA antibodies in sera of dogs naturally infected with Toxocara canis, Ancylostoma caninum, and Dipylidium caninum, although only weakly by IgE. Adult parasite proteins were recognized by antibodies in sera of dogs naturally infected with gastrointestinal helminths. Adult parasite proteins with m.w. of approximately 51, 63, 92, and 209 kDa recognized by IgG could be used for specific diagnosis of canine angiostrongylosis.