The occurrence of subtilase-cytotoxin-encoding genes in environmental Escherichia coli isolated from a Northern California estuary.

The presence of subtilase-cytotoxin-encoding genes was determined in 397 environmental Escherichia coli strains isolated from water, suspended solids, and sediments sampled from different hydrological and environmental conditions in a California estuary. A total of 7 strains (1.76%) were found to harbor subtilase-cytotoxin-encoding genes. Using primers targeting subA only, we generated PCR amplicons from 2 strains; while using primers targeting the 3' end of SubA downstream to the 5' end of SubB, amplicons of 232 bp were generated from 5 additional strains. The 556 bp subA sequences were almost identical to that in the subtilase-cytotoxin-positive strain ED 591 (98%), while subAB sequences of 2 non-Shiga-toxigenic strains revealed 100% similarity with the Shiga-toxigenic E. coli O113:H21 strain 98NK2 that was isolated from an outbreak of hemolytic uremic syndrome. Additionally, the serogroup O113:H21 was present in this collection of environmental E. coli, and it was found to harbor stx2d, hra1 that encodes the heat resistant agglutinin 1, and a subAB sequence similar to that in the non-Shiga-toxigenic E. coli subtilase cytotoxin strain ED 591. To further understand potential health risks posed by strains encoding SubAB, future epidemiological studies should consider screening isolates for subAB regardless of the presence of Shiga-toxin-encoding genes.

Spatial and Temporal Dynamics of Fecal Coliform and Associated with Suspended Solids and Water within Five Northern California Estuaries.

Fecal coliform and associated with suspended solids (SS) and water in five northern California estuaries were studied to document process influences and water quality monitoring biases affecting indicator bacteria concentrations. We collected and analyzed 2371 samples during 10 sampling events for the five studied estuaries. Concentrations during wet-season stormflow conditions were greater than during wet-season base flow and dry-season base flow conditions. Results also document concentration gradients across the length of the studied estuaries and with depth of sample collection. Highest concentrations were associated with shallow samples collected furthest inland. Corresponding decreases occurred the deeper and closer to the estuary mouth a sample was collected. Results also identify direct relationships of wind speed and discharge velocity and indirect relationship of tide stage to indicator bacteria concentrations. Bacteria associated with suspended solids (SS), after conversion to the same units of measurement (mass), were three orders of magnitude greater than in the water fraction. However, the mean proportion contributed by SS to composite water sample concentrations was 8% (SE 0.3) for fecal coliform and 7% (SE 0.3) for . Bacteria from the SS proportion is related to seasonality, tide stage, and discharge velocity that are consistent with mechanisms for entrainment, transport of SS, and reduced particle settling. These results are important for both managing and monitoring these systems by improving sample spatial and temporal context and corresponding bacteria concentration values across the freshwater-saltwater interface.

Giardia duodenalis in feedlot cattle from the central and western United States.

BACKGROUND: Giardia duodenalis is a ubiquitous protozoan parasite that has emerged as a significant opportunistic human pathogen. G. duodenalis may have a deleterious effect on animal growth and performance, therefore its potential as a production limiting organism should not be discounted. We therefore undertook this study to determine management and environmental factors in feedlots that influence the prevalence and environmental load of G. duodenalis cysts in fecal material deposited by feedlot cattle in the central and western United States. RESULTS: Twenty two feedlots from 7 states were included in the study, and up to 240 fecal samples were collected from pen floors of up to 6 pens per feedlot. Giardia duodenalis cysts were identified and counted using direct immunofluorescent microscopy. The estimated overall point prevalence of G. duodenalis was 19.1%, representing feedlots from a wide range of climates and management systems. Pen-level prevalence varied from 0 to 63.3%, with pen-level shedding estimates ranging from 0 to 261,000 cysts/g feces. Higher environmental temperatures, increased animal density, and increased time in the feedlot were associated with a lower prevalence of G. duodenalis. Removing manure before placing a new group of cattle in a pen was associated with a decreased prevalence of G. duodenalis in fecal pats. Using coccidiostats as a feed additive was associated with a higher prevalence of Giardia. CONCLUSION: Management practices could be employed that would limit the probability that feedlot cattle shed G. duodenalis in their feces and therefore potentially limit contamination of their environment.

Environmental load of Cryptosporidium parvum oocysts from cattle manure in feedlots from the central and western United States.

The first step in assessing the risk of water contamination by Cryptosporidium parvum oocysts from feedlot cattle (Bos taurus) production systems is to quantify the number of C. parvum oocysts present in the fecal material deposited by feedlot cattle. Our primary objective for this project was to estimate the daily environmental load of C. parvum oocysts in fecal material deposited by feedlot cattle from across the central and western USA. Our secondary goal was to genotype isolates of C. parvum from feedlot cattle to help facilitate proper identification of mammalian sources of waterborne C. parvum. Based on 5274 fecal samples from 22 feedlots in seven states (California, Washington, Colorado, Oklahoma, Texas, Nebraska, and South Dakota), we estimated a point prevalence of C. parvum of 0.99 to 1.08% in fecal material from feedlot pens from a wide range of climates and a diverse range of feedlot management systems. On average, fresh fecal material from throughout feedlot systems (recent arrivals to nearing slaughter) contained about 1.3 to 3.6 oocysts/g feces, which roughly translates to about 2.8 x 10(4) to 1.4 x 10(5) oocysts/animal per day.

Efficacy of vegetated buffer strips for retaining Cryptosporidium parvum.

Overland and shallow subsurface hydrologic transport of pathogenic Cryptosporidium parvum oocysts from cattle feces into surface drinking water supplies is a major concern on annual grasslands in California's central and southern Sierra Nevada foothills. Soil boxes (0.5 m wide x 1.1 m long x 0.3 m deep) were used to evaluate the ability of grass vegetated buffer strips to retain 2 x 10(8) spiked C. parvum oocysts in 200-g fecal deposits during simulated rainfall intensities of 30 to 47.5 mm/h over 2 h. Buffers were comprised of Ahwahnee sandy loam (coarse-loamy, mixed, active, thermic Mollic Haploxeralfs; 78:18:4 sand to silt to clay ratio; dry bulk density = 1.4 g/cm(3)) set at 5 to 20% land slope, and >/=95% grass cover (grass stubble height = 10 cm; biomass = 900 kg/ha dry weight). Total number of oocysts discharged from each soil box (combined overland and subsurface flow) during the 120-min simulation ranged from 1.5 x 10(6) to 23.9 x 10(6) oocysts. Observed overall mean log(10) reduction of total C. parvum flux per meter of vegetated buffer was 1.44, 1.19, and 1.18 for buffers at 5, 12, and 20% land slope, respectively. Rainfall application rate (mm/h) was strongly associated with oocyst flux from these vegetated buffers, resulting in a decrease of 2 to 4% in the log(10) reduction per meter buffer for every additional mm/h applied to the soil box. These results support the use of strategically placed vegetated buffers as one of several management strategies that can reduce the risk of waterborne C. parvum attributable to extensive cattle grazing on annual grassland watersheds.

Seasonal shedding of multiple Cryptosporidium genotypes in California ground squirrels (Spermophilus beecheyi).

Twelve percent of 853 California ground squirrels (Spermophilus beecheyi) from six different geographic locations in Kern County, Calif., were found to be shedding on average 44,482 oocysts g of feces(-1). The mean annual environmental loading rate of Cryptosporidium oocysts was 57,882 oocysts squirrel(-1) day(-1), with seasonal patterns of fecal shedding ranging from <10,000 oocysts squirrel(-1) day(-1) in fall, winter, and spring to levels of 2 x 10(5) oocysts squirrel(-1) day(-1) in summer. Juveniles were about twice as likely as adult squirrels to be infected and shed higher concentrations of oocysts than adults did, with particularly high levels of infection and shedding being found among juvenile male squirrels. Based on DNA sequencing of a portion of the 18S small-subunit rRNA gene, there existed three genotypes of Cryptosporidium species in these populations of squirrels (Sbey03a, Sbey03b, and Sbey03c; accession numbers AY462231 to AY462233, respectively). These unique DNA sequences were most closely related (96 to 97% homology) to porcine C. parvum (AF115377) and C. wrairi (AF115378). Inoculating BALB/c neonatal mice with up to 10,000 Sbey03b or Sbey03c fresh oocysts from different infected hosts did not produce detectable levels of infection, suggesting that this common genotype shed by California ground squirrels is not infectious for mice and may constitute a new species of Cryptosporidium.