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In vitro anthelmintic effects of Spigelia anthelmia protein fractions against Haemonchus contortus.

Abstract Gastrointestinal nematodes are a significant concern for animal health and well-being, and anthelmintic treatment is mainly performed through the use of chemical products. However, bioactive compounds produced by plants have shown promise for development as novel anthelmintics. The aim of this study is to assess the anthelmintic activity of protein fractions from Spigelia anthelmia on the gastrointestinal nematode Haemonchus contortus. Plant parts were separated into leaves, stems and roots, washed with distilled water, freeze-dried and ground into a fine powder. Protein extraction was performed with sodium phosphate buffer (75 mM, pH 7.0). The extract was fractionated using ammonium sulfate (0-90%) and extensively dialyzed. The resulting fractions were named LPF (leaf protein fraction), SPF (stem protein fraction) and RPF (root protein fraction), and the protein contents and activities of the fractions were analyzed. H. contortus egg hatching (EHA), larval exsheathment inhibition (LEIA) and larval migration inhibition (LMIA) assays were performed. Proteomic analysis was conducted, and high-performance liquid chromatography (HPLC) chromatographic profiles of the fractions were established to identify proteins and possible secondary metabolites. S. anthelmia fractions inhibited H. contortus egg hatching, with LPF having the most potent effects (EC50 0.17 mg mL-1). During LEIA, SPF presented greater efficiency than the other fractions (EC50 0.25 mg mL-1). According to LMIA, the fractions from roots, stems and leaves also reduced the number of larvae, with EC50 values of 0.11, 0.14 and 0.21 mg mL-1, respectively. Protein analysis indicated the presence of plant defense proteins in the S. anthelmia fractions, including protease, protease inhibitor, chitinase and others. Conversely, secondary metabolites were absent in the S. anthemia fractions. These results suggest that S. anthelmia proteins are promising for the control of the gastrointestinal nematode H. contortus.
PMID
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Ovicidal and larvicidal activity in vitro of Spigelia anthelmia Linn. extracts on Haemonchus contortus.

Authors

Mayor MeshTerms
Keywords
Journal Title plos one
Publication Year Start




PMID- 29244856
OWN - NLM
STAT- MEDLINE
DCOM- 20180102
LR  - 20180102
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 12
IP  - 12
DP  - 2017
TI  - In vitro anthelmintic effects of Spigelia anthelmia protein fractions against
      Haemonchus contortus.
PG  - e0189803
LID - 10.1371/journal.pone.0189803 [doi]
AB  - Gastrointestinal nematodes are a significant concern for animal health and
      well-being, and anthelmintic treatment is mainly performed through the use of
      chemical products. However, bioactive compounds produced by plants have shown
      promise for development as novel anthelmintics. The aim of this study is to
      assess the anthelmintic activity of protein fractions from Spigelia anthelmia on 
      the gastrointestinal nematode Haemonchus contortus. Plant parts were separated
      into leaves, stems and roots, washed with distilled water, freeze-dried and
      ground into a fine powder. Protein extraction was performed with sodium phosphate
      buffer (75 mM, pH 7.0). The extract was fractionated using ammonium sulfate
      (0-90%) and extensively dialyzed. The resulting fractions were named LPF (leaf
      protein fraction), SPF (stem protein fraction) and RPF (root protein fraction),
      and the protein contents and activities of the fractions were analyzed. H.
      contortus egg hatching (EHA), larval exsheathment inhibition (LEIA) and larval
      migration inhibition (LMIA) assays were performed. Proteomic analysis was
      conducted, and high-performance liquid chromatography (HPLC) chromatographic
      profiles of the fractions were established to identify proteins and possible
      secondary metabolites. S. anthelmia fractions inhibited H. contortus egg
      hatching, with LPF having the most potent effects (EC50 0.17 mg mL-1). During
      LEIA, SPF presented greater efficiency than the other fractions (EC50 0.25 mg
      mL-1). According to LMIA, the fractions from roots, stems and leaves also reduced
      the number of larvae, with EC50 values of 0.11, 0.14 and 0.21 mg mL-1,
      respectively. Protein analysis indicated the presence of plant defense proteins
      in the S. anthelmia fractions, including protease, protease inhibitor, chitinase 
      and others. Conversely, secondary metabolites were absent in the S. anthemia
      fractions. These results suggest that S. anthelmia proteins are promising for the
      control of the gastrointestinal nematode H. contortus.
FAU - Araujo, Sandra Alves
AU  - Araujo SA
AD  - Laboratory of Plant Biochemistry, Chemical Engineering Course, Centre for Exact
      Sciences and Technology, Federal University of Maranhao, Sao Luiz MA, Brazil.
FAU - Soares, Alexandra Martins Dos Santos
AU  - Soares AMDS
AD  - Laboratory of Plant Biochemistry, Chemical Engineering Course, Centre for Exact
      Sciences and Technology, Federal University of Maranhao, Sao Luiz MA, Brazil.
FAU - Silva, Carolina Rocha
AU  - Silva CR
AD  - Laboratory of Parasite Control, Department of Pathology, Center for Biological
      and Health Sciences, Federal University of Maranhao, Sao Luiz MA, Brazil.
FAU - Almeida Junior, Eduardo Bezerra
AU  - Almeida Junior EB
AD  - Laboratory of Botanical Studies, Department of Biology, Center for Biological and
      Health Sciences, Federal University of Maranhao, Sao Luis MA, Brazil.
FAU - Rocha, Claudia Quintino
AU  - Rocha CQ
AD  - Laboratory of Advanced Studies in Phytomedicines, Department of Chemistry, Centre
      for Exact Sciences and Technology, Federal University of Maranhao, Sao Luis MA,
      Brazil.
FAU - Ferreira, Andre Teixeira da Silva
AU  - Ferreira ATDS
AD  - Laboratory of Toxinology, Oswaldo Cruz Foundation, Rio de Janeiro RJ, Brazil.
FAU - Perales, Jonas
AU  - Perales J
AD  - Laboratory of Toxinology, Oswaldo Cruz Foundation, Rio de Janeiro RJ, Brazil.
FAU - Costa-Junior, Livio M
AU  - Costa-Junior LM
AUID- ORCID: http://orcid.org/0000-0002-1475-049X
AD  - Laboratory of Parasite Control, Department of Pathology, Center for Biological
      and Health Sciences, Federal University of Maranhao, Sao Luiz MA, Brazil.
LA  - eng
PT  - Journal Article
DEP - 20171215
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Anthelmintics)
RN  - 0 (Plant Extracts)
RN  - 0 (Plant Proteins)
SB  - IM
MH  - Animals
MH  - Anthelmintics/chemistry/*therapeutic use
MH  - Chromatography, High Pressure Liquid
MH  - Haemonchiasis/*drug therapy/parasitology/veterinary
MH  - Haemonchus/*drug effects/pathogenicity
MH  - Larva/drug effects/pathogenicity
MH  - Loganiaceae/chemistry
MH  - Plant Extracts/*administration & dosage/chemistry
MH  - Plant Leaves/chemistry
MH  - Plant Proteins/administration & dosage/chemistry
MH  - Proteomics
PMC - PMC5731696
EDAT- 2017/12/16 06:00
MHDA- 2018/01/03 06:00
CRDT- 2017/12/16 06:00
PHST- 2017/03/22 00:00 [received]
PHST- 2017/12/02 00:00 [accepted]
PHST- 2017/12/16 06:00 [entrez]
PHST- 2017/12/16 06:00 [pubmed]
PHST- 2018/01/03 06:00 [medline]
AID - 10.1371/journal.pone.0189803 [doi]
AID - PONE-D-17-11322 [pii]
PST - epublish
SO  - PLoS One. 2017 Dec 15;12(12):e0189803. doi: 10.1371/journal.pone.0189803.
      eCollection 2017.