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Shots might be given weekly or monthly

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doi:10.1111/cts.13704

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Samples were fixed in 10% buffered formalin overnight at 4C, dehydrated with increasing concentrations of ethanol, embedded in paraffin, cut into 5 m sections, and stained with hematoxylin and eosin (HE) or Massons Trichrome Stain Kit (Sigma-Aldrich)