Compositions produced using enteric pathogens and methods of use
The present invention provides compositions including polypeptides having the characteristics of polypeptides expressed by a reference microbe such E. coli or Salmonella . Examples of Salmonella strains that can be used include, for instance, S. enterica serovar Newport, S. enterica serovar Enteritidis, S. enterica serovar Typhimurium, and S. enterica serovar Dublin. Also provided are compositions including polypeptides having a particular molecular weight and a mass fingerprint that includes polypeptide fragments having a particular set of masses, or polypeptides having an amino acid sequence with at least about 95% identity with an amino acid sequence, wherein the polypeptide has seroreactive activity. The present invention also provides methods of making and methods of using such compositions.
1. A method for decreasing intestinal colonization of an animal intestinally colonized by, or at risk of being intestinally colonized by a Salmonella spp., the method comprising parenterally administering an effective amount of a composition to the animal, wherein the composition comprises:
a first isolated iron-regulated polypeptide from Salmonella enterica serovar Enteritidis having a molecular weight of about 92 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the first isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having masses of 728.43 Da±300 parts per million (ppm), 815.38 Da±300 ppm, 888.41 Da±300 ppm, 957.50 Da±300 ppm, 972.50 Da±300 ppm, 986.54 Da±300 ppm, 998.45 Da±300 ppm, 1008.47 Da±300 ppm, 1047.53 Da±300 ppm, 1076.59 Da±300 ppm, 1113.56 Da±300 ppm, 1180.59 Da±300 ppm, 1219.61 Da±300 ppm, 1276.60 Da±300 ppm, 1282.74 Da±300 ppm, 1338.66 Da±300 ppm, 1384.65 Da±300 ppm, 1401.71 Da±300 ppm, 1402.67 Da±300 ppm, 1470.69 Da±300 ppm, 1519.80 Da±300 ppm, 1527.72 Da±300 ppm, 1648.75 Da±300 ppm, 1691.87 Da±300 ppm, 1712.86 Da±300 ppm, 1732.81 Da±300 ppm, 1758.82 Da±300 ppm, 1786.87 Da±300 ppm, 1894.97 Da±300 ppm, 1953.86 Da±300 ppm, 2159.05 Da±300 ppm, 2254.95 Da±300 ppm, 2284.27 Da±300 ppm, 2793.46 Da±300 ppm, and 2881.34 Da±300 ppm;
a second isolated iron-regulated polypeptide from Salmonella enterica serovar Enteritidis having a molecular weight of about 91 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the second isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having masses of 904.51 Da±300 ppm, 924.43 Da±300 ppm, 945.53 Da±300 ppm, 1004.49 Da±300 ppm, 1050.50 Da±300 ppm, 1075.60 Da±300 ppm, 1109.55 Da±300 ppm, 1199.63 Da±300 ppm, 1276.56 Da±300 ppm, 1294.58 Da±300 ppm, 1307.68 Da±300 ppm, 1343.65 Da±300 ppm, 1375.72 Da±300 ppm, 1417.75 Da±300 ppm, 1450.68 Da±300 ppm, 1509.63 Da±300 ppm, 1510.72 Da±300 ppm, 1601.89 Da±300 ppm, 1618.81 Da±300 ppm, 1624.79 Da±300 ppm, 1668.72 Da±300 ppm, 1766.91 Da±300 ppm, 1792.85 Da±300 ppm, 1808.87 Da±300 ppm, 1832.89 Da±300 ppm, 2013.02 Da±300 ppm, 2088.13 Da±300 ppm, 2269.01 Da±300 ppm, 2298.03 Da±300 ppm, 2453.09 Da±300 ppm, 2554.16 Da±300 ppm, and 2572.20 Da±300 ppm;
a third isolated iron-regulated polypeptide from Salmonella enterica serovar Enteritidis having a molecular weight of about 86 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the third isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having masses of 643.37 Da±300 ppm, 872.46 Da±300 ppm, 950.49 Da±300 ppm, 990.55 Da±300 ppm, 1082.61 Da±300 ppm, 1084.57 Da±300 ppm, 1095.49 Da±300 ppm, 1151.68 Da±300 ppm, 1181.55 Da±300 ppm, 1207.58 Da±300 ppm, 1324.75 Da±300 ppm, 1365.74 Da±300 ppm, 1377.66 Da±300 ppm, 1411.78 Da±300 ppm, 1432.77 Da±300 ppm, 1436.74 Da±300 ppm, 1499.71 Da±300 ppm, 1560.86 Da±300 ppm, 1584.74 Da±300 ppm, 1618.77 Da±300 ppm, 1633.84 Da±300 ppm, 1727.83 Da±300 ppm, 1870.95 Da±300 ppm, 1903.94 Da±300 ppm, 1974.95 Da±300 ppm, 1980.96 Da±300 ppm, 1997.06 Da±300 ppm, 2078.00 Da±300 ppm, 2192.94 Da±300 ppm, 2233.14 Da±300 ppm, 2371.13 Da±300 ppm, 2531.24 Da±300 ppm, 2622.42 Da±300 ppm, 2632.20 Da±300 ppm, 3098.47 Da±300 ppm, 3211.45 Da±300 ppm, and 3473.51 Da±300 ppm;
a fourth isolated iron-regulated polypeptide from Salmonella enterica serovar Enteritidis having a molecular weight of about 83 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the fourth isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having masses of 610.29 Da±300 ppm, 628.39 Da±300 ppm, 848.45 Da±300 ppm, 918.45 Da±300 ppm, 1040.60 Da±300 ppm, 1097.62 Da±300 ppm, 1141.60 Da±300 ppm, 1153.63 Da±300 ppm, 1162.50 Da±300 ppm, 1218.66 Da±300 ppm, 1309.63 Da±300 ppm, 1335.71 Da±300 ppm, 1341.66 Da±300 ppm, 1364.60 Da±300 ppm, 1405.76 Da±300 ppm, 1460.70 Da±300 ppm, 1528.70 Da±300 ppm, 1564.76 Da±300 ppm, 1564.76 Da±300 ppm, 1735.86 Da±300 ppm, 1750.86 Da±300 ppm, 1754.83 Da±300 ppm, 1845.91 Da±300 ppm, 1880.96 Da±300 ppm, 1911.98 Da±300 ppm, 1954.02 Da±300 ppm, 2030.93 Da±300 ppm, 2261.06 Da±300 ppm, 2397.12 Da±300 ppm, 2416.14 Da±300 ppm, 2701.36 Da±300 ppm, 2909.36 Da±300 ppm, and 2943.50 Da±300 ppm; and
a fifth polypeptide having a molecular weight of about 78 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the fifth isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having masses of 605.33 Da±300 ppm, 614.38 Da±300 ppm, 616.37 Da±300 ppm, 808.41 Da±300 ppm, 836.42 Da±300 ppm, 989.50 Da±300 ppm, 1060.52 Da±300 ppm, 1063.48 Da±300 ppm, 1140.66 Da±300 ppm, 1158.55 Da±300 ppm, 1177.52 Da±300 ppm, 1210.55 Da±300 ppm, 1314.62 Da±300 ppm, 1329.77 Da±300 ppm, 1345.55 Da±300 ppm, 1493.80 Da±300 ppm, 1526.73 Da±300 ppm, 1570.82 Da±300 ppm, 1649.90 Da±300 ppm, 1654.83 Da±300 ppm, 1740.90 Da±300 ppm, 1744.69 Da±300 ppm, 1750.84 Da±300 ppm, 1792.88 Da±300 ppm, 1814.85 Da±300 ppm, 1906.92 Da±300 ppm, 1952.94 Da±300 ppm, 2242.03 Da±300 ppm, 2538.26 Da±300 ppm, and 2710.17 Da±300 ppm.
2. The method of claim 1 wherein the animal is avian, bovine, caprine, ovine, porcine, bisontine, cervine, equine, a companion animal, or human.
3. The method of claim 2 wherein the avian animal is a chicken or turkey.
4. The method of claim 1 further comprising one or more additional administrations of the composition to the animal.
5. The method of claim 1 wherein the parenteral administration comprises subcutaneous administration.
6. A method of decreasing mortality due to infection by a Salmonella spp. in an animal, the method comprising parenterally administering an effective amount of a composition to the animal, wherein the composition comprises:
a first isolated iron-regulated polypeptide from Salmonella enterica serovar Enteritidis having a molecular weight of about 92 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the first isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having masses of 728.43 Da±300 parts per million (ppm), 815.38 Da±300 ppm, 888.41 Da±300 ppm, 957.50 Da±300 ppm, 972.50 Da±300 ppm, 986.54 Da±300 ppm, 998.45 Da±300 ppm, 1008.47 Da±300 ppm, 1047.53 Da±300 ppm, 1076.59 Da±300 ppm, 1113.56 Da±300 ppm, 1180.59 Da±300 ppm, 1219.61 Da±300 ppm, 1276.60 Da±300 ppm, 1282.74 Da±300 ppm, 1338.66 Da±300 ppm, 1384.65 Da±300 ppm, 1401.71 Da±300 ppm, 1402.67 Da±300 ppm, 1470.69 Da±300 ppm, 1519.80 Da±300 ppm, 1527.72 Da±300 ppm, 1648.75 Da±300 ppm, 1691.87 Da±300 ppm, 1712.86 Da±300 ppm, 1732.81 Da±300 ppm, 1758.82 Da±300 ppm, 1786.87 Da±300 ppm, 1894.97 Da±300 ppm, 1953.86 Da±300 ppm, 2159.05 Da±300 ppm, 2254.95 Da±300 ppm, 2284.27 Da±300 ppm, 2793.46 Da±300 ppm, and 2881.34 Da±300 ppm;
a second isolated iron-regulated polypeptide from Salmonella enterica serovar Enteritidis having a molecular weight of about 91 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the second isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having masses of 904.51 Da±300 ppm, 924.43 Da±300 ppm, 945.53 Da±300 ppm, 1004.49 Da±300 ppm, 1050.50 Da±300 ppm, 1075.60 Da±300 ppm, 1109.55 Da±300 ppm, 1199.63 Da±300 ppm, 1276.56 Da±300 ppm, 1294.58 Da±300 ppm, 1307.68 Da±300 ppm, 1343.65 Da±300 ppm, 1375.72 Da±300 ppm, 1417.75 Da±300 ppm, 1450.68 Da±300 ppm, 1509.63 Da±300 ppm, 1510.72 Da±300 ppm, 1601.89 Da±300 ppm, 1618.81 Da±300 ppm, 1624.79 Da±300 ppm, 1668.72 Da±300 ppm, 1766.91 Da±300 ppm, 1792.85 Da±300 ppm, 1808.87 Da±300 ppm, 1832.89 Da±300 ppm, 2013.02 Da±300 ppm, 2088.13 Da±300 ppm, 2269.01 Da±300 ppm, 2298.03 Da±300 ppm, 2453.09 Da±300 ppm, 2554.16 Da±300 ppm, and 2572.20 Da±300 ppm;
a third isolated iron-regulated polypeptide from Salmonella enterica serovar Enteritidis having a molecular weight of about 86 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the third isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having masses of 643.37 Da±300 ppm, 872.46 Da±300 ppm, 950.49 Da±300 ppm, 990.55 Da±300 ppm, 1082.61 Da±300 ppm, 1084.57 Da±300 ppm, 1095.49 Da±300 ppm, 1151.68 Da±300 ppm, 1181.55 Da±300 ppm, 1207.58 Da±300 ppm, 1324.75 Da±300 ppm, 1365.74 Da±300 ppm, 1377.66 Da±300 ppm, 1411.78 Da±300 ppm, 1432.77 Da±300 ppm, 1436.74 Da±300 ppm, 1499.71 Da±300 ppm, 1560.86 Da±300 ppm, 1584.74 Da±300 ppm, 1618.77 Da±300 ppm, 1633.84 Da±300 ppm, 1727.83 Da±300 ppm, 1870.95 Da±300 ppm, 1903.94 Da±300 ppm, 1974.95 Da±300 ppm, 1980.96 Da±300 ppm, 1997.06 Da±300 ppm, 2078.00 Da±300 ppm, 2192.94 Da±300 ppm, 2233.14 Da±300 ppm, 2371.13 Da±300 ppm, 2531.24 Da±300 ppm, 2622.42 Da±300 ppm, 2632.20 Da±300 ppm, 3098.47 Da±300 ppm, 3211.45 Da±300 ppm, and 3473.51 Da±300 ppm;
a fourth isolated iron-regulated polypeptide from Salmonella enterica serovar Enteritidis having a molecular weight of about 83 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the fourth isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having masses of 610.29 Da±300 ppm, 628.39 Da±300 ppm, 848.45 Da±300 ppm, 918.45 Da±300 ppm, 1040.60 Da±300 ppm, 1097.62 Da±300 ppm, 1141.60 Da±300 ppm, 1153.63 Da±300 ppm, 1162.50 Da±300 ppm, 1218.66 Da±300 ppm, 1309.63 Da±300 ppm, 1335.71 Da±300 ppm, 1341.66 Da±300 ppm, 1364.60 Da±300 ppm, 1405.76 Da±300 ppm, 1460.70 Da±300 ppm, 1528.70 Da±300 ppm, 1564.76 Da±300 ppm, 1564.76 Da±300 ppm, 1735.86 Da±300 ppm, 1750.86 Da±300 ppm, 1754.83 Da±300 ppm, 1845.91 Da±300 ppm, 1880.96 Da±300 ppm, 1911.98 Da±300 ppm, 1954.02 Da±300 ppm, 2030.93 Da±300 ppm, 2261.06 Da±300 ppm, 2397.12 Da±300 ppm, 2416.14 Da±300 ppm, 2701.36 Da±300 ppm, 2909.36 Da±300 ppm, and 2943.50 Da±300 ppm; and
a fifth polypeptide having a molecular weight of about 78 kDa as measured following electrophoresis on an SDS-polyacrylamide gel, wherein the fifth isolated iron-regulated polypeptide, if digested with trypsin, produces polypeptide fragments having masses of 605.33 Da±300 ppm, 614.38 Da±300 ppm, 616.37 Da±300 ppm, 808.41 Da±300 ppm, 836.42 Da±300 ppm, 989.50 Da±300 ppm, 1060.52 Da±300 ppm, 1063.48 Da±300 ppm, 1140.66 Da±300 ppm, 1158.55 Da±300 ppm, 1177.52 Da±300 ppm, 1210.55 Da±300 ppm, 1314.62 Da±300 ppm, 1329.77 Da±300 ppm, 1345.55 Da±300 ppm, 1493.80 Da±300 ppm, 1526.73 Da±300 ppm, 1570.82 Da±300 ppm, 1649.90 Da±300 ppm, 1654.83 Da±300 ppm, 1740.90 Da±300 ppm, 1744.69 Da±300 ppm, 1750.84 Da±300 ppm, 1792.88 Da±300 ppm, 1814.85 Da±300 ppm, 1906.92 Da±300 ppm, 1952.94 Da±300 ppm, 2242.03 Da±300 ppm, 2538.26 Da±300 ppm, and 2710.17 Da±300 ppm.
7. The method of claim 6 wherein the animal is avian, bovine, caprine, ovine, porcine, bisontine, cervine, equine, a companion animal, or human.
8. The method of claim 7 wherein the avian animal is a chicken or turkey.
9. The method of claim 6 further comprising one or more additional administrations of the composition to the animal.
10. The method of claim 6 wherein the parenteral administration comprises subcutaneous administration.