[GaryPassamonte]

John- The coloring may have been done in the 1860s or later. I still think the bouquet is the key. Why would someone add a bouquet to a photograph if the players weren't women? It just doesn't add up. As I said prior, we will probably never know with certainty the gender of these players. However, all factors considered, I believe there is a better chance these players are women than boys.
Joshua- Thanks. I will try to contact her.

After researching the history of women's clothing in the 1850s and 1860s, we can't rule out that the trousers may in fact be bloomers. There was a feminist movement at this time and the wearing of trousers or bloomers was done. Any woman "bold" enough to play baseball in the 1860s may have "bold" enough to wear trousers.