P' westermani Kerbert, 1878 Best known species, human parasite that can undergo development in 16 di

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Paragonimus
Pre-adult
Egg 80-11048-60µ
Encysted metacercaria 300- 400µ
Adult 7.5-12 4-61-3mm (l w 21)

• P. westermani (Kerbert, 1878) - Best known
  species, human parasite that can undergo
  development in gt16 different snail species and 50
  crustacean species.
• Status of prevalence and host range in India-
  not well documented.

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Distribution of human paragonimiasis
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Distribution in India
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Paragonimus species India
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Life Cycle

• Infective stage metacercaria
• Infective mode eating raw fresh watercrabs and
  crayfish with metacercariae
• Infective route by mouth
• Site of inhabitation lungs
• Intermediate hosts 1st snail 2ndcrab,
  crayfish
• Reservoir hosts carnivores (tiger, lion, wolf,
  fox, dog, leopard, cat etc.)
• Life span 5-6 years

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Molecular Taxonomy
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The ribosomal DNA gene cluster
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Bioinformatic Tools
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Materials Methods
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DNA amplification and sequencing
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Molecular Phylogenetic Analysis

• Only unique sequences were used in tree
  construction.
• ITS sequences entered in MEGA for phylogenetic
  trees construction.
• Tree building methods- Maximum Parsimony,
  Neighbor-Joining, UPGMA,
• Minimum
  Evolution.
• Branch support given using 1000 bootstrap
  replicates in MEGA

Bayesian Phylogenetic Analysis
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Motif identification, testing and validation

• Sequences motifs identified from aligned
  sequences of the data set using PRATT software.
• Motifs were expressed using DNA alphabet
  (A,T,C,G) in PROSITE language.
• Validation of motifs were performed for each
  species using a PATTERN MATCHING web
  application.

Evaluation through BLAST analysis
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Predicted ITS2 RNA secondary structures and
analyses
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Results
PCR amplification and analysis
1b
PCR products of Paragonimus metacercaria DNA
using primer set 3S - A28 for ITS2
Product size 500 bp Final reaction volume
50µl 1.6 agarose gel electrophoresis Marker Ø
x 174 DNA/ HaeIII Digest
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NJ tree
MP tree
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BLAST outputs of Paragonimus ITS sequence motifs
against NCBI GenBank database
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Discussion
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Conclusion
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Acknowledgements

• DST, DBT, CSIR (GOI)- for Travel Fellowship for
  InCoB2009.
• DIT Project to VT.
• AICOPTAX programme (MoEF, GOI) to VT
• DBT Project to VT AC.
• DSA programme (UGC-SAP) in Zoology
• UPE (Biosciences) programme in School of Life
  Sciences, NEHU, Shillong.
• Co-ordinator, Bioinformatics Centre, NEHU.