Maria das Graças C. Pereira

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

ABSTRACT 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 × 105 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.

Prevalence and associated risk factors for Giardia lamblia infection among children hospitalized for diarrhea in Goiânia, Goiás State, Brazil

The objective of this study was to determine the prevalence and to identify risk factors associated with Giardia lamblia infection in diarrheic children hospitalized for diarrhea in Goiânia, State of Goiás, Brazil. A cross-sectional study was conducted and a comprehensive questionnaire was administered to the childs primary custodian. Fixed effects logistic regression was used to determine the association between infection status for G. lamblia and host, sociodemographic, environmental and zoonotic risk factors. A total of 445 fecal samples were collected and processed by the DFA methodology, and G. lamblia cysts were present in the feces of 44 diarrheic children (9.9%). A variety of factors were found to be associated with giardiasis in these population: age of children (OR, 1.18; 90% CI, 1.0 - 1.36; p = 0.052), number of children in the household (OR 1.45; 90% CI, 1.13 - 1.86; p = 0.015), number of cats in the household (OR, 1.26; 90% CI, 1.03 -1.53; p = 0.059), food hygiene (OR, 2.9; 90% CI, 1.34 - 6.43; p = 0.024), day-care centers attendance (OR, 2.3; 90% CI, 1.20 - 4.36; p = 0.034), living on a rural farm within the past six months prior hospitalization (OR, 5.4; CI 90%, 1.5 - 20.1; p = 0.03) and the number of household adults (OR, 0.59; 90% CI, 0.42 - 0.83; p = 0.012). Such factors appropriately managed may help to reduce the annual incidence of this protozoal infection in the studied population.

Climate and On-Farm Risk Factors Associated with Giardia duodenalis Cysts in Storm Runoff from California Coastal Dairies

ABSTRACT Climatic factors and on-farm management practices were evaluated for their association with the concentrations (cyst/liter) and instantaneous loads (cysts/second) of Giardia duodenalis in storm-based runoff from dairy lots and other high-cattle-use areas on five coastal California farms over two storm seasons. Direct fluorescent antibody analysis was used to quantitate cysts in 350 storm runoff samples. G. duodenalis was detected on all five dairy farms, with fluxes of 1 to 14,000 cysts/liter observed in 16% of samples. Cysts were detected in 41% of runoff samples collected near cattle less than 2 months old, compared to 10% of runoff samples collected near cattle over 6 months old. Furthermore, the concentrations and instantaneous loads of cysts were ≥65 and ≥79 times greater, respectively, in runoff from sites housing young calves than in sites housing other age classes of animals. Factors associated with environmental loading of G. duodenalis included cattle age, cattle stocking number, and precipitation but not lot area, land slope, or cattle density. Vegetated buffer strips were found to significantly reduce waterborne cysts in storm runoff: each additional meter of vegetated buffer placed below high-cattle-use areas was associated with reductions in the concentration and instantaneous load of cysts by factors of 0.86 and 0.79 (−0.07 and −0.10 log10/m), respectively. Straw mulch, seed application, scraping of manure, and cattle exclusion did not significantly affect the concentration or load of G. duodenalis cysts. The study findings suggest that vegetated buffer strips, especially when placed near dairy calf areas, should help reduce the environmental loading of these fecal protozoa discharging from dairy farms.

Efficacy of natural grassland buffers for removal of Cryptosporidium parvum in rangeland runoff

Our goal for this project was to estimate the retention efficiency of natural grassland buffers for Cryptosporidium parvum. Three sets of 16 plots (2.0 by 3.0 m) were established at 5, 20, and 35% slopes. Within each set of 16 plots, residual dry vegetation matter treatments of 225, 560, and 900 kg/ha were implemented, along with a noncut control averaging 4,500 kg/ ha. Buffer width treatments were implemented by placing cattle fecal material containing known loads of C. parvum 0.1, 1.1, or 2.1 m up-slope of the runoff collector. Grassland buffers of 1.1 and 2.1 m generated 3.2- to 8.8-log and 3.6- to 8.8-log retention of C. parvum, respectively, across the range of residual dry vegetation matter, land slope, rainfall, and runoff conditions examined during this project. Buffers with an increased percent land slope exhibited improved the retention efficiencies, whereas buffers experiencing larger maximum annual runoff events exhibited reduced retention efficiencies. Water-quality data from the 0.1-m-wide buffer plots (effectively no buffer) demonstrated that the majority of C. parvum oocysts (98 to 99.999%) were retained in the fecal matrix for the duration of the storm season, irrespective of the presence of a vegetated buffer. In conclusion, these results support the assertion that grassland buffers are an effective method for reducing animal agricultural inputs of waterborne C. parvum into drinking and irrigation water supplies.

Cryptosporidium rubeyi n. sp. (Apicomplexa: Cryptosporidiidae) in multiple Spermophilus ground squirrel species

Previously we reported the unique Cryptosporidium sp. “c” genotype (e.g., Sbey03c, Sbey05c, Sbld05c, Sltl05c) from three species of Spermophilus ground squirrel (Spermophilus beecheyi, Spermophilus beldingi, Spermophilus lateralis) located throughout California, USA. This follow-up work characterizes the morphology and animal infectivity of this novel genotype as the final step in proposing it as a new species of Cryptosporidium. Analysis of sequences of 18S rRNA, actin, and HSP70 genes of additional Cryptosporidium isolates from recently sampled California ground squirrels (S. beecheyi) confirms the presence of the unique Sbey-c genotype in S. beecheyi. Phylogenetic and BLAST analysis indicates that the c-genotype in Spermophilus ground squirrels is distinct from Cryptosporidium species/genotypes from other host species currently available in GenBank. We propose to name this c-genotype found in Spermophilus ground squirrels as Cryptosporidium rubeyi n. sp. The mean size of C. rubeyi n. sp. oocysts is 4.67 (4.4–5.0) μm × 4.34 (4.0–5.0) μm, with a length/width index of 1.08 (n = 220). Oocysts of C. rubeyi n. sp. are not infectious to neonatal BALB/c mice and Holstein calves. GenBank accession numbers for C. rubeyi n. sp. are DQ295012, AY462233, and KM010224 for the 18S rRNA gene, KM010227 for the actin gene, and KM010229 for the HSP70 gene.

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

BackgroundGiardia 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.ResultsTwenty 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.ConclusionManagement 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.

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.