Efficacy of European starling control to reduce Salmonella enterica contamination in a concentrated animal feeding operation in the Texas panhandle.

BACKGROUND: European starlings (Sturnus vulgaris) are an invasive bird species known to cause damage to plant and animal agriculture. New evidence suggests starlings may also contribute to the maintenance and spread of diseases within livestock facilities. Identifying and mitigating the risk pathways that contribute to disease in livestock is necessary to reduce production losses and contamination of human food products. To better understand the impact starlings have on disease transmission to cattle we assessed the efficacy of starling control as a tool to reduce Salmonella enterica within a concentrated animal feeding operation. We matched a large facility, slated for operational control using DRC-1339 (3-chloro-4-methylaniline hydrochloride, also 3-chloro p-toluidine hydrochloride, 3-chloro-4-methylaniline), with a comparable reference facility that was not controlling birds. In both facilities, we sampled cattle feed, cattle water and cattle feces for S. enterica before and after starling control operations. RESULTS: Within the starling-controlled CAFO, detections of S. enterica contamination disappeared from feed bunks and substantially declined within water troughs following starling control operations. Within the reference facility, detections of S. enterica contamination increased substantially within feed bunks and water troughs. Starling control was not observed to reduce prevalence of S. enterica in the cattle herd. Following starling control operations, herd prevalence of S. enterica increased on the reference facility but herd prevalence of S. enterica on the starling-controlled CAFO stayed at pretreatment levels. CONCLUSIONS: Within the starling-controlled facility detections of S. enterica disappeared from feed bunks and substantially declined within water troughs following control operations. Since cattle feed and water are obvious routes for the ingestion of S. enterica, starling control shows promise as a tool to help livestock producers manage disease. Yet, we do not believe starling control should be used as a stand alone tool to reduce S. enterica infections. Rather starling control could be used as part of a comprehensive disease management plan for concentrated animal feeding operations.

Experimental exposure of swallow bugs (Hemiptera: Cimicidae) to West Nile virus.

The cliff swallow (Petrochelidon pyrrhonota, Vieillot) could potentially play an important role in the transmission of West Nile virus (WNV) because of its breeding ecology, reservoir competence status, and potentially high natural exposure rates. These birds nest in colonies and their nests are occupied year round by swallow bugs (Oeciacus vicarius, Hovarth), hematophagus ectoparasites that feed primarily on cliff swallows. Swallow bugs are most likely exposed to WNV while feeding on infectious blood of cliff swallows and, thus, if competent vectors, could contribute to initiation and maintenance of seasonal WNV transmission. In addition, swallow bugs remain within nests year round and, therefore, if persistently infected and competent vectors, they could provide an overwintering mechanism for WNV. We tested the hypothesis that swallow bugs become infected with WNV through direct abdominal inoculation or ingestion of infectious blood meals. We observed that swallow bugs did not maintain or amplify WNV, and infectious virus titers within bugs declined over 15 d postexposure. These results suggest that swallow bugs may not be competent vectors of WNV, and therefore are unlikely to play a significant role in transmission.

Serologic evidence of avian influenza (H4N6) exposure in a wild-caught raccoon.

Growing concerns about avian influenza, and its effect on agriculture and human health, have highlighted the need to understand the role of wildlife in maintaining and spreading the virus. We surveyed the wildlife inhabiting a poultry farm with recent H3N6 and H4N6 avian influenza virus exposure in Pennsylvania, U.S.A. One raccoon (Procyon lotor) tested positive for H4N6 antibodies. This is the first recorded incident of avian influenza exposure in a wild raccoon. We suggest that raccoons may play a role in the transmission of avian influenza viruses and in compromising biosecurity efforts at poultry operations.

Influenza exposure in United States feral swine populations.

Swine play an important role in the disease ecology of influenza. Having cellular receptors in common with birds and humans, swine provide opportunities for mixed infections and potential for genetic reassortment between avian, human, and porcine influenza. Feral swine populations are rapidly expanding in both numbers and range and are increasingly coming into contact with waterfowl, humans, and agricultural operations. In this study, over 875 feral swine were sampled from six states across the United States for serologic evidence of exposure to influenza. In Oklahoma, Florida, and Missouri, USA, no seropositive feral swine were detected. Seropositive swine were detected in California, Mississippi, and Texas, USA. Antibody prevalences in these states were 1% in Mississippi, 5% in California, and 14.4% in Texas. All seropositive swine were exposed to H3N2 subtype, the predominant subtype currently circulating in domestic swine. The only exceptions were in San Saba County, Texas, where of the 15 seropositive samples, four were positive for H1N1 and seven for both H1N1 and H3N2. In Texas, there was large geographical and temporal variation in antibody prevalence and no obvious connection to domestic swine operations. No evidence of exposure to avian influenza in feral swine was uncovered. From these results it is apparent that influenza in feral swine poses a risk primarily to swine production operations. However, because feral swine share habitat with waterfowl, prey on and scavenge dead and dying birds, are highly mobile, and are increasingly coming into contact with humans, the potential for these animals to become infected with avian or human influenza in addition to swine influenza is a distinct possibility.

Serologic evidence of West Nile virus exposure in North American mesopredators.

Sera from 936 mammalian mesopredators (Virginia opossums, gray foxes, striped skunks, hooded skunks, raccoons, a bobcat, and a red fox) were collected during 2003 and 2004 in California, Arizona, Texas, Louisiana, Ohio, and Wyoming and screened for flavivirus-specific antibodies by an epitope-blocking enzyme-linked immunosorbent assay (blocking ELISA). Serum samples positive for antibodies against flaviviruses were screened for West Nile virus (WNV)-specific antibodies by blocking ELISA and selectively confirmed with plaque-reduction neutralization tests. High prevalence rates were observed in raccoons (45.6%) and striped skunks (62.9%). The high WNV antibody prevalence noted in mesopredators, their peridomestic tendencies, and their overall pervasiveness make these species potentially useful sentinels for monitoring flaviviruses in defined areas.

Fox squirrel (Sciurus niger) associations with West Nile virus.

Tree squirrels (Sciurus spp.) have been recently shown to be commonly exposed to West Nile virus (WNV). Many characteristics of WNV infections in tree squirrels are unknown. To better understand WNV associations in fox squirrels (S. niger), we conducted mark-recapture sampling (N = 72) and radio telemetry to study the longitudinal seroprevalence, seroconversions, and ectoparasites of these animals during 2005-2006 in northern Colorado. Five seroconversions were documented during this study. The majority of seroconversions occurred during the late summer/fall months. However, one seroconversion was documented over the time period of February to late March 2005. Fleas (Orchopeas howardi) were tested for WNV RNA using real-time PCR techniques. No WNV RNA positive fleas (N = 33) were detected. In addition, urine samples (N = 17) opportunistically collected from fox squirrels were negative for WNV RNA. Results indicate that seroconversions can be observed in fox squirrels during low WNV transmission years.

Mosquito landing rates on nesting American robins (Turdus migratorius).

We measured mosquito landing rates on adult and nestling American robins at nests with infrared cameras in Washington, D.C., and Maryland, United States. Mosquitoes landed on nesting robins almost exclusively between dusk and dawn. The mean number of mosquito landings per night was higher for adults (123.3 +/- SE 32.8) than nestlings (37.26 +/- 14.8). The fraction of mosquitoes landing at a nest on nestlings increased with decreases in adult brooding. Oral swabs from nestlings at these and 13 other robin, Gray catbird, and house finch nests were negative for West Nile virus (WNV). These results show that landing rates were higher on adults and that parental brooding reduces the landing rates of mosquitoes on nestlings.

Experimental infection of fox squirrels (Sciurus niger) with West Nile virus.

Tree squirrels (Sciurus spp.) have exhibited high seroprevalence rates, suggesting that they are commonly exposed to West Nile virus (WNV). Many characteristics of WNV infections in tree squirrels, such as the durations and levels of viremia, remain unknown. To better understand WNV infections in fox squirrels (S. niger), we subcutaneously inoculated fourteen fox squirrels with WNV. Peak viremias ranged from 10(4.00) plaque-forming units (PFU)/mL of serum on day 2 post-infection (DPI) to 10(4.98) PFU/mL on 3 DPI, although viremias varied between individuals. Oral secretions of some fox squirrels were positive for WNV viral RNA, occasionally to moderate levels (10(3.2) PFU equivalent/swab). WNV PFU equivalents in organs were low or undetectable on 12 DPI; gross and histologic lesions were rare. The viremia profiles of fox squirrels indicate that they could serve as amplifying hosts in nature. In addition, viral RNA in the oral cavity and feces indicate that this species could contribute to alternative WNV transmission in suburban communities.

West Nile virus antibody prevalence in red-winged blackbirds (Agelaius phoeniceus) from North Dakota, USA (2003-2004).

This study was designed to explore the role that red-winged blackbirds (Agelaius phoeniceus) may have played in disseminating West Nile virus (WNV) across the United States. Using enzyme-linked immunosorbent assays designed to detect WNV antibodies in avian species we were able to determine the WNV antibody prevalence in a cohort of red-winged blackbirds in central North Dakota in 2003 and 2004. The peak WNV antibody prevalence was 22.0% in August of 2003 and 18.3% in July of 2004. The results of this study suggest that red-winged blackbird migratory populations may be an important viral dispersal mechanism with the ability to spread arboviruses such as WNV across the United States.

Prevalence of shiga toxin-producing Escherichia coli and Salmonella enterica in rock pigeons captured in Fort Collins, Colorado.

The potential role of rock pigeons (Columba livia) in the epidemiology of shiga toxin-producing Escherichia coli (STEC) and Salmonella enterica is unclear. Our objective was to determine the prevalence of STEC and S. enterica in pigeons at urban and dairy settings as a function of season. Prevalence of STEC and S. enterica was estimated by bacteriologic culture of cloacal swabs collected from pigeons trapped at urban and dairy locations in and around Fort Collins, Colorado from January to November 2003. Presumptive E. coli isolates were tested for the presence of virulence genes SLT-1, SLT-2, eae, hlyA, K1, CNF-1, CNF-2, and LT using polymerase chain reaction. Shiga toxins were not isolated from any of 406 samples from pigeons, but virulence genes typically associated with disease in humans were identified in isolates from 7.9% (95% CI: 5.5% to 10.9%) of captured pigeons. S. enterica were detected in 3.2% of 277 samples from pigeons, with all positive samples originating from dairy locations (nine of 106 [8.5%]; 95% CI: 4.0-15.5%). The results suggest that although pigeons may acquire S. enterica from cattle and play a role in recirculation and persistence of the microorganism at dairies, pigeons are not important carriers of STEC.

Susceptibility of greater sage-grouse to experimental infection with West Nile virus.

Populations of greater sage-grouse (Centrocercus urophasianus) have declined 45-80% in North America since 1950. Although much of this decline has been attributed to habitat loss, recent field studies have indicated that West Nile virus (WNV) has had a significant negative impact on local populations of grouse. We confirm the susceptibility of greater sage-grouse to WNV infection in laboratory experimental studies. Grouse were challenged by subcutaneous injection of WNV (10(3.2) plaque-forming units [PFUs]). All grouse died within 6 days of infection. The Kaplan-Meier estimate for 50% survival was 4.5 days. Mean peak viremia for nonvaccinated birds was 10(6.4) PFUs/ml (+/-10(0.2) PFUs/ml, standard error of the mean [SEM]). Virus was shed cloacally and orally. Four of the five vaccinated grouse died, but survival time was increased (50% survival=9.5 days), with 1 grouse surviving to the end-point of the experiment (14 days) with no signs of illness. Mean peak viremia for the vaccinated birds was 10(2.3) PFUs/ml (+/-10(0.6) PFUs/ml, SEM). Two birds cleared the virus from their blood before death or euthanasia. These data emphasize the high susceptibility of greater sage-grouse to infection with WNV.

West Nile virus serosurveillance in Iowa white-tailed deer (1999-2003).

Sera from white-tailed deer (Odocoileus virginianus) were collected in Iowa during the winter months (1999-2003), 2 years before and after West Nile virus (WNV) was first reported in Iowa (2001), and were analyzed for antibodies to WNV. Samples from 1999 to 2001 were antibody negative by blocking enzyme-linked immunosorbent assay (bELISA) and plaque reduction neutralization test (PRNT(90)). Prevalence derived from bELISA (2002, 12.7%; 2003, 11.2%) and WNV PRNT(90) (2002, 7.9%; 2003, 8.5%) assays were similar. All samples were negative for antibodies against St. Louis encephalitis virus as determined by PRNT(90). Antibodies to flaviviruses were detected by indirect enzyme-linked immunosorbent assay (iELISA) prior to the first WNV cases reported in Iowa (1999-2001) with prevalence ranging from 2.2% to 3.2%, suggesting the circulation of an additional undescribed flavivirus prior to the introduction of WNV into the area. Flavivirus prevalence as determined by iELISA increased in 2002 and 2003 (23.3% and 31.9%, respectively). The increase in prevalence exceeded estimates of WNV prevalence, suggesting that conditions favored general flavivirus transmission (including WNV) during the 2002-2003 epizootic. These data indicate that serologic analysis of deer sera collected from hunter harvests may prove useful for surveillance and evidence of local transmission of WNV and other pathogens and identify white-tailed deer as a species for further studies for host competency.

Serologic evidence of exposure of wild mammals to flaviviruses in the central and eastern United States.

Serosurveys were conducted to obtain flavivirus and West Nile virus (WNV) seroprevalence data from mammals. Sera from 513 small- and medium-sized mammals collected during late summer and fall 2003 from Colorado, Louisiana, New York, Ohio, and Pennsylvania were screened for flavivirus-specific antibodies. Sera samples containing antibody to flaviviruses were screened for WNV-specific antibodies by epitope-blocking enzyme-linked immunosorbent assays and confirmed with plaque reduction neutralization tests. Prevalence of WNV antibodies among study sites ranged from 0% to 42.8% among the mammal communities sampled. High prevalence rates for WNV were noted among raccoons (100%, with a very small sample size, N = 2), Virginia opossums (50.0%), fox squirrels (49.1%), and eastern gray squirrels (48.3%). The high WNV antibody prevalence noted for tree squirrels, the peri-domestic tendencies of several of these species, and their ease of observation could make these species useful sentinels for monitoring WNV activity within urban communities.

Baseline characteristics and the effect of selenium supplementation on cancer incidence in a randomized clinical trial: a summary report of the Nutritional Prevention of Cancer Trial.

The Nutritional Prevention of Cancer Trial was a randomized, clinical trial designed to evaluate the efficacy of selenium as selenized yeast (200 microg daily) in preventing the recurrence of nonmelanoma skin cancer among 1312 residents of the Eastern United States. Original secondary analyses through December 31, 1993 showed striking inverse associations between treatment and the incidence of total [hazard ratio (HR) = 0.61, 95% confidence interval (CI) = 0.46-0.82], lung, prostate, and colorectal cancer and total cancer mortality. This report presents results through February 1, 1996, the end of blinded treatment. Effect modification by baseline characteristics is also evaluated. The effects of treatment overall and within subgroups of baseline age, gender, smoking status, and plasma selenium were examined using incidence rate ratios and Cox proportional hazards models. Selenium supplementation reduced total (HR = 0.75, 95% CI = 0.58-0.97) and prostate (HR = 0.48, 95% CI = 0.28-0.80) cancer incidence but was not significantly associated with lung (HR = 0.74, 95% CI = 0.44-1.24) and colorectal (HR = 0.46, 95% CI = 0.21-1.02) cancer incidence. The effects of treatment on other site-specific cancers are also described. The protective effect of selenium was confined to males (HR = 0.67, 95% CI = 0.50-0.89) and was most pronounced in former smokers. Participants with baseline plasma selenium concentrations in the lowest two tertiles (<121.6 ng/ml) experienced reductions in total cancer incidence, whereas those in the highest tertile showed an elevated incidence (HR = 1.20, 95% CI = 0.77-1.86). The Nutritional Prevention of Cancer trial continues to show a protective effect of selenium on cancer incidence, although not all site-specific cancers exhibited a reduction in incidence. This treatment effect was restricted to males and to those with lower baseline plasma selenium concentrations.

Selenium supplementation and lung cancer incidence: an update of the nutritional prevention of cancer trial.

Interest in the chemopreventive effects of the trace element selenium has spanned the past three decades. Of >100 studies that have investigated the effects of selenium in carcinogen-exposed animals, two-thirds have observed a reduction in tumor incidence and/or preneoplastic endpoints (G. F. Combs and S. B. Combs, The Role of Selenium in Nutrition Chapter 10, pp. 413-462. San Diego, CA: Academic Press, 1986, and B. H. Patterson and O. A. Levander, Cancer Epidemiol. Biomark. Prev., 6: 63-69, 1997). The Nutritional Prevention of Cancer Trial, a randomized clinical trial reported by Clark et al. (L. C. Clark et al., JAMA, 276: 1957-1963, 1996), showed as a secondary end point, a statistically significant decrease in lung cancer incidence with selenium supplementation. The adjusted hazard ratio (HR) was 0.56 [95% confidence interval (CI), 0.31-1.01; P = 0.05]. These results were based on active follow-up of 1312 participants. This reanalysis used an extended Nutritional Prevention of Cancer Trial participant follow-up through the end of the blinded clinical trial on February 1, 1996. The additional 3 years added 8 cases to the selenium-treated group and 4 cases to the placebo group, and increased follow-up to 7.9 years. The relative risk of 0.70 (95% CI, 0.40-1.21; P = 0.18) is not statistically significant. Whereas the overall adjusted HR is not significant (HR = 0.74; 95% CI, 0.44-1.24; P = 0.26), and the HR for current and former smokers was not significant, the trend is toward a reduction in risk of incident lung cancer with selenium supplementation. In a subgroup analysis there was a nominally significant HR among subjects with baseline plasma selenium in the lowest tertile (HR = 0.42; 95% CI, 0.18-0.96; P = 0.04). The analysis for the middle and highest tertiles of baseline showed HRs of 0.91 and 1.25. The current reanalysis indicates that selenium supplementation did not significantly decrease lung cancer incidence in the full population, but a significant decrease among individuals with low baseline selenium concentrations was observed.

Safety issues regarding colonic cleansing for diagnostic and surgical procedures.

There are various methods available to cleanse the colon in preparation for diagnostic and surgical procedures. The popular options are diet and cathartic regimens, gut lavage and phosphate preparations. Each method has its own unique characteristics and safety profile. Diet and cathartic regimens are based on traditional methods of colonoscopy preparation and remain an acceptable and safe alternative for patients unwilling or unable to tolerate other bowel preparations. Gut lavage methods involve ingestion of 2-4L of osmotically balanced solutions containing polyethylene glycol, which have been shown to be safe and effective for colon cleansing, including for special patient populations with cardiac, renal or hepatic dysfunction. Phosphate preparations have also been shown to be safe and effective for colon cleansing and are generally better tolerated than counterpart gut lavage solutions. However, this method has safety concerns for some patients with cardiac, renal and hepatic dysfunctions.

Interaction Between Sensory and Postingestional Repellents in Starlings: Methyl Anthranilate and Sucrose.

Ingestion of concentrated sucrose solutions causes sickness in sucrase-deficient birds. As a result, some suggest that sucrose may represent an environmentally safe avian repellent. In the present experiments, we compared the repellency of sucrose to that of methyl anthranilate (MA), a known avian repellent. We also tested mixtures of sucrose and MA to investigate whether repellency could be enhanced, relative to sucrose or MA presented alone. The results showed that the MA was strongly avoided in both drinking and feeding trials. Conversely, only high sucrose concentrations were avoided, and only in drinking trials. No combination of MA and sucrose was as effective as MA alone. We conclude that sucrose is not sufficiently aversive to serve as an avian repellent in the field.

Detection of low-molecular-weight proteins in urine by dipsticks.

BACKGROUND: Testing of urines with dipsticks for proteinuria, glycosuria, etc., is common practice. A deficiency with currently available dipsticks is their lack of chemical sensitivity and underestimation of low-molecular-weight proteins such as light chains. METHODS: We experimented with a number of dyes that gave an easily recognized color change on dipsticks for various low-molecular-weight proteins such as alpha-1-glycoprotein, alpha-1- and beta-2-microglobulin, and kappa and lambda light chains. We were successful in formulating a dye for impregnating dipsticks that gave a color change with low-molecular-weight proteins. RESULTS: Most dipsticks will measure proteins down to about 1 g/l. Our composite of two dyes (described here as the "TPR" dipsticks) gave reproducible results for protein concentrations of >/=300 mg/l, and detected low-molecular proteins. The TPR reagent is resistant to interferences from many compounds; also, the protein results are not altered in a given urine at a pH between 5 and 8. CONCLUSIONS: We have developed a dipstick that detects low-molecular-weight proteins. The dipsticks are easy to use and are suitable for outpatient or point-of-care testing. The precision of the dipsticks is satisfactory and is only marginally lower than quantitative spectrophotometric methods using pyrogallol red (PYR).

Effect of biologically active plants used as netst material and the derived benefit to starling nestlings.

The European starling Sturnus vulgaris preferentially incorporates fresh sprigs of particular plant species for use as nesting material. Chemicals found in these plants may act to reduce pathogen and ectoparasite populations normally found in nest environments. The present experiments were performed to test this Nest Protection Hypothesis. In the fild, we experimentally determined that wild carrot Daucus carota, a plant species preferred as nest material, effectively reduced the number of hematophagous mites found within nests relative to control nests without green vegetation. Chicks from nests containing wild carrot had higher levels of blood hemoglobin than chicks from control nests. However, there were no differences in weight or feather development. In the laboratory, we found that wild carrot and fleabane, Erigeron philadelphicus, (also preferred by starlings as nest material) substantially reduced the emergence of feeding instars of mites, while garlic mustard, Alliaria officinalis, (commonly available but not preferred) had little effect on the emergence of mites. We infer that preferred plant material may act to inhibit feeding or otherwise delay reproduction of mites, thereby reducing risk of anemia to developing nestlings.

A report of high-dose selenium supplementation: response and toxicities.

Concerns about the toxicity of selenium has limited the doses used in chemoprevention. Based on previous studies, intakes of 400 microg/day and plasma selenium of 1000 ng/ml (Dietary Reference Intakes, Academy Press, New York, 2000, p. 384) were established as the no observed adverse effect level (NOAEL). This investigation summarizes the plasma response and toxicity reports from 24 men with biopsy-proven prostate cancer who were randomized to either 1600 or 3200 microg/day of selenized yeast as part of a controlled clinical trial testing selenium as a chemopreventive agent for prostate cancer progression. Subjects were on these doses for averages of almost 12 months. Plasma selenium levels were monitored throughout the course of follow-up. Symptoms of selenium toxicity were assessed by patient interview with specific questions regarding breath, hair and nail changes. Several liver and kidney function tests and hematology were measured at 6-month intervals. 8 subjects were randomized to the 1600 microg/day and 16 to the 3200 microg/day group. The mean plasma selenium levels achieved with supplementation were 492.2 ng/ml (SD = 188.3) and 639.7 ng/ml (SD = 490.7) for the 1600 and 3200 microg/ day doses, respectively. The 3200 microg/day group reported more selenium-related side effects. Blood chemistry and hematology results were all within normal limits for both treatment groups. More subjects on 3200 microg/day reported symptoms of selenium toxicity; however, these reports did not correspond to peaks in plasma selenium levels. We observed no obvious selenium-related serious toxicities. As selenium is used in more chemoprevention and therapeutic settings, additional information on selenium species, sequestration of selenium in specific organs, excretion, and toxicities is needed.