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Item A molecular phylogenetic study of the caecal fluke of poultry, Postharmostomum commutatum (= P. gallinum) (Trematoda: Brachylaimidae)(Parasitology Research, 2018-12) Valadão, Marisa C; Silva, Beatriz C. M.; López-Hernández, Danimar; Araújo, Jackson V.; Locke, Sean A.; Pinto, Hudson A.,Postharmostomum commutatum (Dietz, 1858), a parasite of the caeca of poultry, has been reported from many different parts of the world. Despite its importance, there are no molecular sequences available and its phylogenetic position is unknown in relation to other members of Brachylaimoidea, a group in which taxonomic confusion reigns. Here, morphological and molecular techniques were used to study digeneans from the caeca of free-range chickens found naturally infected in the municipality of Viçosa, state of Minas Gerais, Brazil, between August 2017 and May 2018. The specimens were identified as P. commutatum, with Postharmostomum gallinum Witenberg, 1923 herein considered a junior synonym. Sequences obtained for the 28S, ITS2, and cox-1 genes were compared with sequences available from other species of Brachylaimoidea. Phylogenetic analysis of the three markers indicates P. commutatum formed an isolated lineage from other brachylaimoids, supporting the distinct status of the genus. The topology of phylogenetic trees obtained suggests that the morphology-based classification of families of Brachylaimoidea is artificial and new rearrangements of some genera or creation of new families may be necessary. The sequences newly obtained here will be useful for testing the cosmopolitan distribution of P. commutatum.Item In vitro activity of a serine protease from Monacrosporium thaumasium fungus against first-stage larvae of Angiostrongylus vasorum(Parasitology Research, 2012-01-10) Braga, Fabio R.; Araújo, Jackson V.; Soares, Filippe E. F.; Lima, Walter dos Santos; Mozer, Lanuze R.; Queiróz, José H.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. Mg2+ and Zn2+ 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 L1, 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.Item Efficacy of Duddingtonia flagrans and Arthrobotrys robusta in controlling sheep parasitic gastroenteritis(Parasitology Research, 2010-05-17) Braga, Fabio R.; Araújo, Jackson V.; Silva, Bruna F.; Mauad, Juliana R. Carrijo; Campos, Artur K.; Amarante, Alessandro F. T.The aim of this study was to evaluate the efficacy of formulations of sodium alginate matrix (pellets) of the nematode predatory fungi, Duddingtonia flagrans (AC001 isolate) and Arthrobotrys robusta (I-31 isolate), in the biological control of sheep gastrointestinal nematode infections. Thirty young Bergamacia ewes were allocated into three groups: In group 1 (control), the animals received 2 g/10 kg of live weight (l.w.) of pellets without fungus; in group 2, each animal received 2 g/10 kg of l.w. of pellets of D. flagrans (0.2 g of fungus/10 kg l.w.); and in group 3, each animal received 2 g/10 kg of l.w. of pellets of A. robusta (0.2 g of fungus/10 kg l.w.). The animals of each group were kept separately under rotational grazing. Pellets, with or without fungi, were mixed with 1 kg animal food and administered twice a week for 6 months. There was no significant difference in mean live weight and packed cell volume among groups (P > 0.05). Mean nematode fecal egg counts (FEC) did not significantly differ between the control and the remaining groups, except in one or two collections, when FEC was higher in the control group than in group 2 and group 3, respectively. The group that received A. robusta pellets needed less salvage anthelmintic treatments. Haemonchus contortus was the predominant species recovered from tracer lambs. The nematophagous fungi, D. flagrans and A. robusta, did not provide satisfactory results in the prophylaxis of parasitic gastroenteritis in sheep, under the conditions of the present study.Item Destruction of Anoplocephala perfoliata eggs by the nematophagous fungus Pochonia chlamydosporia(Journal of Equine Veterinary Science, 2010-12) Silva, André R.; Araújo, Jackson V.; Braga, Fábio R.; Alves, Camila D. F.; Ribeiro Filho, José DantasThe in vitro effect of an isolate of the nematophagous fungus Pochonia chlamydosporia (VC1) on the eggs of Anoplocephala perfoliata was evaluated. The eggs were morphologically analyzed for their integrity using light microscopy (10× objectives), plated on 9.0-cm diameter petri dishes containing 2% WA culture medium with and without fungal isolate (control), grown for 10 days, and 10 replicates were prepared per group. In all, 1000 eggs of A perfoliata were plated on petri dishes containing 2% water agar culture medium with (VC1) and without the fungal isolate (control). After 3, 5, 7, and 10 days, approximately 100 eggs were removed from each plate and classified on the basis of the following parameters: without alteration; type 1, lytic effect without morphological damage to eggshell; type 2, lytic effect with morphological alteration of embryo and eggshell; and type 3, lytic effect with morphological alteration of embryo and eggshell, in addition to hyphal penetration and internal egg colonization and destruction. The P chlamydosporia fungus demonstrated ovicidal activity (P < .05) on the eggs of A perfoliata in the studied intervals presenting type 3 effects of 35%, 42.5%, 53.83%, and 71.17% for the intervals 3, 5, 7, and 10 days, respectively. P chlamydosporia is a potential biological control agent for the eggs of A perfoliata.Item Ovicidal activity of seven Pochonia chlamydosporia fungal isolates on Ascaris suum eggs(Tropical Animal Health and Production, 2010-11-19) Ferreira, Sebastião R.; Araújo, Jackson V.; Braga, Fabio R.; Araujo, Juliana M.; Carvalho, Rogério O.; Silva, André R.; Frassy, Luiza N.; Freitas, Leandro G.The ovicidal effect of the nematophagous fungus Pochonia chlamydosporia on eggs of Ascaris suum was tested under laboratory conditions. A. suum eggs were plated on 2% water–agar with seven fungal isolates (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) and control without fungus. After 5, 7, 10, 14, 15 and 21 days of incubation, approximately 100 eggs were removed from the plates and classified according to the following parameters: type 1, biochemical and physiological effect without morphological damage to the eggshell, type 2, lytic effect with morphological alteration of the eggshell and embryo and type 3, lytic effect with morphological alteration of eggshell and embryo showing hyphal penetration and internal egg colonization. The isolates effectively destroyed A. suum eggs and all types of effects were observed during the experiment. There was no variation in ovicidal capacity (type 3 effect) among the isolates (p > 0.05) throughout the experiment. After 21 days, isolate 5 showed the highest percentages of type 3 effect (58.33%). The results indicated that P. chlamydosporia (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) can destroy A. suum eggs and is, therefore, a potential biological control agent of nematodes.Item In vitro predatory activity of the fungi Duddingtonia flagrans, Monacrosporium thaumasium, Monacrosporium sinense and Arthrobotrys robusta on Ancylostoma ceylanicum third-stage larvae(Veterinary Microbiology, 2010-05-03) Braga, Fabio R.; Silva, André R.; Carvalho, Rogério O.; Araújo, Jackson V.; Guimarães, Pedro Henrique G.; Fujiwara, Ricardo T.; Frassy, Luiza N.The potential role of companion animals as reservoirs for zoonotic diseases has been recognised as a significant public health problem worldwide. Ancylostoma ceylanicum is the only ancylostomatidae species known for infecting human beings. This article aimed to compare the predatory capacity of predatory fungi isolates Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34), Monacrosporium sinense (SF53) and Arthrobotrys robusta (I31) on A. ceylanicum infectious larvae (L3) in a 2% water–agar plate. There was no predatory capacity variation among the fungi tested (P > 0.05) over the 7-day period experimental assay. When compared to the control (without fungi), there was a significant reduction (P < 0.05) of 95.6%, 85.1%, 87.4% and 90.2% on the A. ceylanicum L3 mean recovered from treatments with isolates AC001, NF34, SF53 and I31, respectively. Regarding linear regression coefficients, negative values were noted for treatments, therefore indicating A. ceylanicum non-predated larvae reduction over 7 days. In this work, all predatory fungi isolates were efficient at capturing and destroying in vitro the A. ceylanicum L3; therefore being able to be used as biological controllers of such nematode.Item Predatory activity of the nematophagous fungus Duddingtonia flagrans on horse cyathostomin infective larvae(Tropical Animal Health and Production, 2010-03-07) Braga, Fabio R.; Araújo, Jackson V.; Silva, André. R.; Carvalho, Rogério O.; Araujo, Juliana M.; Ferreira, Sebastião R.; Benjamin, Laércio A.This work was performed to determine the predatory capacity in vitro of the nematophagous fungus Duddingtonia flagrans (isolate AC001) on cyathostomin infective larvae of horse (L3). The experimental assay was carried out on plates with 2% water-agar (2% WA). In the treated group, each plate contained 1.000 L3 and 1.000 conidia of the fungus. The control group without fungus only contained 1.000 L3 in the plates. Ten random fields (4 mm diameter) were examined per plate of treated and control groups, every 24 h for seven days under an optical microscope (10× and 40× objective lens) for non-predated L3 counts. After 7 days, the non-predated L3 were recovered from the Petri dishes using the Baermann method. The interaction there was a significant reduction (p < 0.01) of 93.64% in the cyathostomin L3 recovered. The results showed that the D. flagrans is a potential candidate to the biological control of horse cyathostomin L3.Item In vitro predatory activity of nematophagous fungi and after passing through gastrointestinal tract of equine on infective larvae of Strongyloides westeri(Parasitology Research, 2010-04-06) Araujo, Juliana M.; Araújo, Jackson V.; Braga, Fabio R.; Carvalho, Rogério O.Three isolates of predator fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34), and Arthrobotrys robusta (I-31) were assessed in in vitro test regarding the capacity of prey infective larvae (L3) Strongyloides westeri. Compared to control, without fungus, there was a significant decrease (P < 0.01) of 80.4%, 67.9%, and 72.8% in means of infective larvae S. westeri recovered from treatments with isolates AC001, NF34, and I-31, respectively. All tested isolates were efficient in the capture of S. westeri (P > 0.01) in vitro test. Linear regression coefficients of treated and control groups were −0.21 for control, −0.32 for D. flagrans, −0.34 for M. thaumasium, and −0.22 for A. robusta. In the following, isolates AC001 and NF34 were assessed in vivo regarding the capacity of supporting the passage through equine gastrointestinal tract without loss of ability of preying infective larvae S. westeri. Fungal isolates survived the passage and were efficient in preying L3 since the first 12 h of collection (P < 0.01) in relation to the control group (without fungus). Compared to control, there was a significant decrease (P < 0.01) of 76.4% and 76.7% (12 h), 86.4% and 85.9% (24 h), 88.3% and 87.7% (48 h), and 89.9% and 87.2% (72 h) in means of infective larvae S. westeri recovered from treatments with isolates AC001 and NF34, respectively. Linear regression coefficients of L3 of recovered S. westeri regarding the collections due to time were 1.93 for control, −3.52 for AC001, and −2.64 for NF34. Fungi D. flagrans and M. thaumasium (NF34) have demonstrated to be promising for use in the biological control of equine parasite S. westeri.Item Biological control of Fasciola hepatica eggs with the Pochonia chlamydosporia fungus after passing through the cattle gastrointestinal tract(Parasitology Research, 2011-07-20) Dias, Anderson S.; Araújo, Jackson V.; Braga, Fábio R.; Araujo, Juliana M.; Puppin, André C.; Fernandes, Fernanda M.; Ramos, Rafael F.; Bertonceli, Raul M.; Silva, Renata G. da; Perboni, Wilber R.Fasciolosis is a disease caused by Fasciola hepatica responsible for causing significant losses in livestock. This study aimed to evaluate the Pochonia chlamydosporia fungus (isolate VC1) on F. hepatica eggs after passing through the cattle gastrointestinal tract. For this evaluation, 1 g pellet was given in sodium alginate matrix per kilogram live weight containing 25% of fungal mycelium from isolate VC1 per animal. Twelve animals were used, six treated and six untreated (control). Some stool samples were collected from the groups of treated and control animals, at the times of 12, 18, 24, 48, 72, and 96 h after the pellets' administration. Then, from each stool sample of treated and control groups, 2 g was placed in a Petri dish of 9 cm in diameter, containing 2% water–agar and 1,000 eggs of F. hepatica. It was observed that the fungus was effective in preying upon the eggs in the samples recovered at all of the schedules starting at 12 h. Furthermore, differences were observed (p < 0.01) in the destruction of eggs in the Petri dishes in the treated group compared with the control group. The ovicidal effect was observed after 7 days of interaction. The ovicidal P. chlamydosporia fungus was effective in destroying F. hepatica eggs; therefore, it is suggested that this fungus could be employed as agent for the control of helminth eggs.Item Predatory capability of the nematophagous fungus Arthrobotrys robusta preserved in silica gel on infecting larvae of Haemonchus contortus(Tropical Animal Health and Production, 2014-02-08) Braga, Fabio R.; Carvalho, Rogério O.; Silva, André R.; Araújo, Jackson V.; Frassy, Luiza N.; Lafisca, Andrea; Soares, Filippe E. F.Biological control of gastrointestinal nematodiasis in ruminants is an alternative to reduce the number of infective larvae. The fungal isolates predatory activity preservation is a basic requirement for the success of this control type. The aim of this work is to evaluate the predatory capacity of the fungus Arthrobotrys robusta (isolate I-31), preserved on silica gel on infective larvae of Haemonchus contortus under laboratory conditions on 2 % water agar (2 % WA). In this essay, A. robusta storage on silica gel showed successful predatory activity on H. contortus L3 larvae (p < 0.01) compared to the control group. Nematophagous fungi were not observed in the control group during the experiment. There was a significant reduction (p < 0.01) of 73.84 % in the means of H. contortus (L3) recovered from treatment with isolate I-31 compared to the control without fungi. Results indicate that A. robusta (I-31) could survive stored on silica gel for at least 7 years and keep its predatory activity on H. contortus (L3).