Eight concentration and purification methods were evaluated to determine percentages of recovery of Cryptosporidium parvumoocysts from calf feces. The NaCl flotation method generally resulted in the highest percentages of recovery. Based on the percentages of recovery, the amounts of fecal debris in the final oocyst preparations, the relatively short processing time (<3 h), and the low expense, the NaCl flotation method was chosen for further evaluation. Extraction efficiency was evaluated by using oocyst concentrations of 25, 50, 102, 103, 104, and 105oocysts g of bovine feces−1. The percentages of recovery ranged from 10.8% (25 oocysts g−1) to 17.0% (104 oocysts g−1) (r 2 = 0.996). A conservative estimate of the detection limit for bovine feces is ca. 30 oocysts g of feces−1. Percentages of recovery were determined for six different types of animal feces (cow, horse, pig, sheep, deer, and chicken feces) at a single oocyst concentration (104 oocysts g−1). The percentages of recovery were highest for bovine feces (17.0%) and lowest for chicken feces (3.2%). Percentages of recovery were determined for bovine manure after 3 to 7 days of storage. The percentages of recovery ranged from 1.9 to 3.5% depending on the oocyst concentration, the time of storage, and the dispersing solution. The percentages of oocyst recovery from soils were evaluated by using different flotation solutions (NaCl, cold sucrose, ZnSO4), different dispersing solutions (Triton X-100, Tween 80, Tris plus Tween 80), different dispersion techniques (magnetic stirring, sonication, blending), and different dispersion times (5, 15, and 30 min). Twenty-five-gram soil samples were used to reduce the spatial variability. The highest percentages of recovery were obtained when we used 50 mM Tris–0.5% Tween 80 as the dispersing solution, dispersion for 15 min by stirring, and saturated NaCl as the flotation solution. The percentages of oocyst recovery from freshly spiked sandy loam, silty clay loam, and clay loam soils were ca. 12 to 18, 8, and 6%, respectively. The theoretical detection limits were ca. 1 to 2 oocysts g of soil−1 depending on the soil type. The percentages of recovery without dispersant (distilled H2O or phosphate-buffered saline) were less than 0.1%, which indicated that oocysts adhere to soil particles. The percentages of recovery decreased with storage time, although the addition of dispersant (Tris-Tween 80) before storage appeared to partially prevent adhesion. These data indicate that the NaCl flotation method is suitable for routine detection and enumeration of oocysts from feces, manures, soils, or soil-manure mixtures.

The transmission of disease in ground water is a topic of great concern to government agencies, ground water specialists, and the general public. The purpose of this study was to compare the temporal variability in storm flow of fecal coliform bacteria densities and Cryptosporidium parvum oocyst densities in agriculturally impacted karst ground water. Cryptosporidium parvum oocyst densities ranged from 0 to 1,050 oocysts/1, and mean storm densities ranged from 3.5 to 156.8 oocysts/1. Fecal coliform densities ranged from less than 1 CFU/100ml to more than 40,000 CFU/100ml, and geometric mean storm densities ranged from 1.7 CFU/100ml to more than 7,000 CFU/100ml. Fecal coliform densities correlated well with flow during storms, but Cryptosporidium oocyst densities exhibited a great deal of sample to sample variability and were not correlated with flow. Fecal coliform densities did not correlate positively with Cryptosporidium oocyst densities. Fecal coliform densities were greatest at storm peaks, when sediment loads were also greatest. Multiple transport mechanisms for fecal coliform bacteria and C. parvum oocysts may necessitate various agricultural land management and livestock health maintenance practices to control movement of pathogens to karst ground water.

No abstract

The objective of this work was to assess the effect of dilute bovine manure (1.0% and 0.1%) versus that of no manure on attachment and subsequent detachment of Cryptosporidium parvum oocysts to soil. Manure enhanced the attachment of oocysts to soil particles; the maximum attachment was observed with 0.1% manure. Oocyst attachment was partially reversible; maximum detachment was observed with dilute manure. These results indicate that oocyst attachment to soil is substantially affected by bovine manure in a complex manner and should have implications for how oocysts may be transported through or over soils.

No abstract