Multilocus Sequence Subtyping and Genetic Structure of <em>Cryptosporidium muris</em> and <em>Cryptosporidium andersoni</em>

<div><p>In this study, nine <em>C. muris</em> and 43 <em>C. andersoni</em> isolates from various animals in China were subtyped by a multilocus sequence typing (MLST) tool. DNA sequence analyses showed the presence of 1–2 subtypes of <em>C. muris</em> and 2–6 subtypes of <em>C. andersoni</em> at each of the four loci (MS1, MS2, MS3, and MS16), nine of which represented new subtypes. Altogether, two <em>C. muris</em> and 10 <em>C. andersoni</em> MLST subtypes were detected. Linkage disequilibrium analysis indicated although the overall population structure of the two parasites was clonal, the Chinese <em>C. andersoni</em> in cattle has an epidemic structure. Three and two clusters were produced in the <em>C. muris</em> and <em>C. andersoni</em> populations by Structure 2.3.3 analysis, with Chinese <em>C. muris</em> and <em>C. andersoni</em> substructures differing from other countries. Thus, this study suggested the prevalence of <em>C. andersoni</em> in China is not attributed to the introduction of dairy cattle. More studies involving more genetic loci and systematic sampling are needed to better elucidate the population genetic structure of <em>C. muris</em> and <em>C. andersoni</em> in the world and the genetic basis for the difference in host specificity among the two most common gastric parasites.</p> </div

Analysis of linkage disequilibrium in <i>C. andersoni</i> and <i>C. muris</i> populations.

<p><b>Note</b>: <i>V</i>_D = the pairwise variance, <i>L = </i>95% critical value; ^awith the same MLST type as one individual; the data of the non-Chinese isolates came from a recently published paper <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0043782#pone.0043782-Feng1" target="_blank">[³¹]</a>.</p

The nature of minisatellite repeats at four genetic loci.

<p>The nature of minisatellite repeats at four genetic loci.</p

Population structure inferred by Bayesian clustering using multilocus subtype information.

<p>A, <i>Cryptosporidium muris</i>; B, <i>Cryptosporidium andersoni</i>. Each individual is shown as a thin vertical line, which is partitioned into <i>K</i> colored components representing estimated membership fractions in <i>K</i> genetic clusters, and the geographic locations are at the bottom. The pie charts show the distribution of genetic clusters in different countries and various animals. JP = Japan; PE = Peru; Ken = Kenya; EG = Egypt; CS = Czech Republic; CN = China; B = beef cattle; C = bactrian camel; D = dairy cattle; H = hamster; S = sheep.</p

Isolates used in this study and their subtype identity at the four minisatellite loci.

<p>Isolates used in this study and their subtype identity at the four minisatellite loci.</p

Genetic diversity of <i>C. andersoni</i> and <i>C. muris</i> DNA sequences.

<p>Genetic diversity of <i>C. andersoni</i> and <i>C. muris</i> DNA sequences.</p

Additional file 2 of Neospora caninum inhibits tumor development by activating the immune response and destroying tumor cells in a B16F10 melanoma model

Additional file 2: Figure S1. Evolution of body weight in tumor-bearing mice (n = 5) treated with PBS or N. caninum tachyzoites

Additional file 3 of Neospora caninum inhibits tumor development by activating the immune response and destroying tumor cells in a B16F10 melanoma model

Additional file 3: Figure S2. Visualization of N. caninum-GFP tachyzoites in B16F10 cells

Additional file 1 of Neospora caninum inhibits tumor development by activating the immune response and destroying tumor cells in a B16F10 melanoma model

Additional file 1: Table S1. The primers used for RT-PCR