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

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          Abstract

          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 L 1 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 L 1 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 L 1.

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          Most cited references13

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          Angiostrongylus vasorum (Baillet, 1866) Kamensky, 1905: emergence of third-stage larvae from infected Biomphalaria glabrata snails.

          Biomphalaria glabrata snails were experimentally infected with Angiostrongylus vasorum first-stage larvae and divided into four groups of 30 snails. To assess the shedding of third-stage larvae (L3), the snails were maintained under different stimuli: group 1 60 W light bulb for 24 h, group 2 37 degrees C water bath for 24 h, group 3 room temperature (23-25 degrees C) for 24 h, Group 4 room temperature (23-25 degrees C) for up to 15 days. After 24 h, a total of 512 A. vasorum L3, alive and active, were released by snails from group 1, while 2,446 L3 were released from group 2 and five L3 from group 3. After 15 days, snails from group 4 released a total of 44 L3. To evaluate the infectivity of A. vasorum L3, two mongrel dogs were successfully infected with L3 released by snails from groups 1 and 2, confirming that the infection of dogs with A. vasorum L3 was possible, independently of ingestion of the mollusk intermediate host. The results shown in these experiments suggest that angiostrongylosis could be directly transmitted to the definitive hosts, with implications for the parasite's life cycle.
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            Larval output of infected and re-infected dogs with Angiostrongylus vasorum (Baillet, 1866) Kamensky, 1905.

            Canine angiostrongylosis is a nematode infection in domestic dogs and wild canids. A natural infection in a domestic dog frequently leads to pneumonia, loss of physical performance, coughing, anemia, cardiac insufficiency, pulmonary fibrosis and death. The main diagnostic method is based on the finding of Angiostrongylus vasorum first-stage larvae (L1) in infected dog feces. With this objective, 11 experimentally exposed to 100 third-stage larvae (L3) per kilogram of body weight (mean = 885.45 L3/animal; S.E. = 77.7). The animals were monitored for 300 days post-single-infection (PI) and the quantity of L1 output measured. Our results showed an irregular excretion of L1 and a variation in the pre-patent period (33-76 days) and the number of L1 excreted by individual animals (1-1261 L1/g). After 300 days PI, five dogs were exposed a second time and monitored for 300 days post-re-infection (PRI) (=600 days PI). The quantity of L1 output demonstrated that double exposed dogs also presented an irregular excretion of L1 but a smaller variation in the number of L1 excreted by individual animals (4-550 L1/g).
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              Angiostrongylus vasorum (Baillet, 1866) Nematoda: Prostostrongylidae, em cães de Minas Gerais, Brasil

              Foi identificado Angiostrongylus vasorum (Baillet, 1866) colhido da artéria pulmonar de dois cães (Canis familiaris) procedentes do município de Caratinga, Estado de Minas Gerais, Brasil. É apresentada a descrição morfológicas do parasita. Esta é a primeira referência desse parasita no Estado de Minas Gerais.
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                Author and article information

                Contributors
                filippeufv@yahoo.com.br
                jqueiroz@ufv.br
                fabioribeirobraga@hotmail.com
                wlima@icb.ufmg.br
                ttonini1@gmail.com
                jvictor@ufv.br
                Journal
                BMC Res Notes
                BMC Res Notes
                BMC Research Notes
                BioMed Central (London )
                1756-0500
                18 November 2014
                18 November 2014
                2014
                : 7
                : 1
                : 811
                Affiliations
                [ ]Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, MG Brasil
                [ ]Departamento de Veterinária, Universidade Federal de Viçosa, Viçosa, MG Brasil
                [ ]Universidade Vila Velha, Vila Velha, ES Brasil
                [ ]Departamento de Parasitologia Animal, Universidade Federal de Minas Gerais, Belo-Horizonte, MG Brasil
                Article
                3328
                10.1186/1756-0500-7-811
                4242478
                25406419
                01d73035-d1cd-4ae9-9cb7-a5afd0971869
                © de Freitas Soares et al.; licensee BioMed Central Ltd. 2014

                This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 22 April 2014
                : 31 October 2014
                Categories
                Short Report
                Custom metadata
                © The Author(s) 2014

                Medicine
                nematophagous fungi,angiostrongylus vasorum,protease
                Medicine
                nematophagous fungi, angiostrongylus vasorum, protease

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