Intranasal dexmedetomidine in healthy beagles: An echocardiographic and pharmacokinetic/pharmacodynamic study.

The aim of this study was to compare the sedative and cardiovascular effects of dexmedetomidine (DEX) administrated via intranasal (IN) and intramuscular (IM) routes. This masked, randomised, crossover study used six male beagle dogs, receiving 0.02 mg/kg dexmedetomidine either IN (DEX-IN) or IM (DEX-IM), and an equal volume of saline by the alternative route. Dexmedetomidine plasma concentration was measured before (TB) and at time points (T) 2, 5, 10, 15, 30, 45, 60, 90 and 120 min after drug administration (T0). Physiological variables, sedation scores and sedation times were recorded until recovery. Echocardiography was performed at TB and between T20-T40. Repeated data over time were analysed using a Scheirer-Ray-Hare test. Other data were compared using a Wilcoxon or Student's t test. Times from T0 to sternal position and from lateral to sternal position were longer for DEX-IN than DEX-IM (P = 0.018 and P = 0.009, respectively). Time from sternal to standing position was shorter for DEX-IN than DEX-IM (P = 0.03). Dexmedetomidine plasma concentrations were significantly lower for DEX-IN than DEX-IM from T10 to T120. Heart rate was significantly lower for DEX-IM than DEX-IN from T5 to T20. Echocardiography showed a decrease in systolic function and calculated cardiac output in DEX-IM as compared to baseline. The DEX concentration-sedation curve for DEX-IN as compared to DEX-IM was leftward shifted, whereas the IN and IM DEX concentration-variation-in-heart-rate curves overlapped. Although reaching lower plasma concentrations, IN dexmedetomidine produced similar sedation to IM dexmedetomidine without affecting cardiovascular function.

Description, characterization, and evaluation of an online social networking community: the American Cancer Society's Cancer Survivors Network®.

PURPOSE: To describe (a) the conceptualization, purpose, and features of The American Cancer Society's Cancer Survivors Network® (CSN; http://csn.cancer.org ), (b) the ongoing two-phase evaluation process of CSN, and (c) the characteristics of CSN members. METHODS: An online opt-in self-report survey of CSN members (N = 4762) was conducted and digital metrics of site use were collected. RESULTS: Annually, CSN attracts over 3.6 million unique users from over 200 countries/territories. Most commonly used site features are discussion boards (81.1%), the search function (63.8%), and the member resource library (50.2%). The survey sample is mostly female (69.6%), non-Hispanic white (84.1%), and self-identified as a cancer survivor (49.8%), or both cancer survivor and cancer caregiver (31.9%). A larger number of survey respondents reported head and neck cancer (12.5%), relative to cancer incidence/prevalence data. CONCLUSIONS: The volume of CSN traffic suggests high demand among cancer survivors and caregivers for informational and/or emotional support from other cancer survivors and caregivers. CSN may be particularly beneficial for individuals with rare cancers. Furthermore, this study documents a group of individuals whose cancer experience is multifaceted (e.g., survivors became caregivers or vice versa), and for whom CSN has the capacity to provide support at multiple points during their cancer experiences. IMPLICATIONS FOR CANCER SURVIVORS: CSN is a free, internet-based social networking site available to all cancer survivors and caregivers, worldwide. Evaluation of the site is ongoing and will be used to inform improvements to usability, reach, recruitment, retention, and potential health impact(s) of this valuable resource.

Arterial oxygen tension and pulmonary ventilation in horses placed in the Anderson Sling suspension system after a period of lateral recumbency and anaesthetised with constant rate infusions of romifidine and ketamine.

REASONS FOR PERFORMING STUDY: Some controversy exists over whether or not horses' recovery and cardiopulmonary function are affected by suspension in slings. OBJECTIVES: To measure arterial oxygen tension and pulmonary ventilation in anaesthetised horses placed in a standing position in an Anderson Sling (AS) after a period of right lateral recumbency (RLR). STUDY DESIGN: Randomised crossover experimental study. METHODS: Six Standardbred horses were anaesthetised twice. Catheters were inserted into the right jugular vein and the left carotid artery. After premedication with romifidine, anaesthesia was induced with diazepam and ketamine. Following 50 min in RLR, horses were maintained in either RLR or AS for an additional 60 min through to recovery. Anaesthesia was maintained i.v. with a constant rate infusion of romifidine and ketamine. Heart rate, respiratory rate, mean arterial pressure, expiratory tidal volume, minute volumes and end tidal CO2 were monitored continuously. Venous and arterial bloods were sampled for lactate concentration, creatine kinase activity and blood gas analysis before premedication, after induction, every 20 min for 100 min, as soon as the horse was standing (TR), and 24 h later. The data were averaged within 2 anaesthetic periods: P1, 0-20 min; and P2, 40-100 min. RESULTS: During P2, horses in the RLR group had lower arterial oxygen tension (P = 0.001), higher alveolar-arterial oxygen tension gradient (P = 0.005), higher respiratory rate (P = 0.04) and higher minute volumes (P = 0.04) than horses in the AS group. Arterial CO2 tension and mean arterial pressure increased in the AS group during P2 (P = 0.01 and 0.02 respectively). The recoveries were judged better in the AS group than in the RLR group (P = 0.01). During TR, lactate were higher in the RLR group than in the AS group (P = 0.007). Creatine kinase activities were higher in the AS group at 24 h vs. TR (P = 0.02). CONCLUSIONS: Anderson Sling suspension after a period of recumbency improves cardiopulmonary function and recovery quality in horses and therefore might be considered for use to recover hypoxic horses.

Blood glucose, acid-base and electrolyte changes during loading doses of alpha2-adrenergic agonists followed by constant rate infusions in horses.

The aim of the present study was to investigate changes in blood glucose concentration ([Glu]B), acid-base status and electrolyte concentrations during constant rate infusions (CRI) of two alpha2-adrenergic agonists in seven horses treated in a blinded, randomised, crossover design with xylazine or romifidine. An intravenous (IV) bolus of xylazine (1mg/kg) or romifidine (80 µg/kg) was administered followed by an IV CRI of xylazine (0.69 mg/kg/h) or romifidine (30 µg/kg/h) for 2h. Blood samples were collected from the pulmonary artery before and after loading doses, during the CRI, and for 1h after discontinuing drugs. Blood glucose, base excess (BE), pH, partial pressure of carbon dioxide (Pv¯CO2), strong ion difference (SIDest) and bicarbonate concentration ( [Formula: see text] ) increased significantly during the CRI with both alpha2-adrenergic agonists. Chloride concentration ([Cl(-)]B) and anion-gap (AG) decreased significantly compared to baseline. The decrease in sodium concentration ([Na(+)]B) was only significant with xylazine. From 1h after starting the CRI onwards, [Glu]B was significantly higher with romifidine compared to xylazine. Except [Glu]B, SIDest, and Pv¯CO2, all variables returned to normal values 1h after discontinuing xylazine. After stopping romifidine, all variables except pH remained altered for at least 1h. It was concluded that loading doses of alpha2-adrenergic agonists followed by CRIs produce [Glu]B, acid-base and electrolyte changes. The clinical significance of the reported changes remains to be investigated and absolute values should be interpreted with caution, as fluid boli were used for cardiac output measurements, but may become important during prolonged infusion and in critically ill patients.

A novel minimal invasive closed chest myocardial ischaemia reperfusion model in rhesus monkeys (Macaca mulatta): improved stability of cardiorespiratory parameters.

The aim of this study was to report the cardiorespiratory events observed during coronary artery occlusion and reperfusion in a minimally invasive closed chest myocardial occlusion-reperfusion model in rhesus monkeys. We hypothesized that a minimally invasive technique may lead to fewer cardiac arrhythmias and complications. Eight male rhesus macaques 10-15 kg and 10-15 years old were sedated with ketamine (2 mg/kg), midazolam (1.3 mg/kg), atropine (0.01 mg/kg) and buprenorphine 0.02 mg/kg intramuscularly. Etomidate 1-2 mg/kg was injected intravenously to allow tracheal intubation. Anaesthesia was maintained with isoflurane. Pulse oximetry, electrocardiogram (ECG), heart rate, mean arterial blood pressure (MAP), inspired isoflurane fractions (F(I)ISO) and core temperature were recorded every 10 min. The coronary artery occlusion was induced by a balloon-tipped catheter advanced via the femoral artery into the left anterior descending artery and inflated to completely occlude the vessel for 20-50 min (IT) before reperfusion. Sequences of elevated ST segment, QRS complex prolongation, ventricular premature complexes and ventricular fibrillation were observed with a lower incidence than previously described in the literature. IT was (min: 17; max: 50) min long. F(I)ISO was lower than the minimal alveolar concentration in these species. Hypotension (MAP < 70 mmHg) and hypothermia (T°C < 36°C) were observed in all macaques. This minimally invasive closed chest model was successful in providing better cardiorespiratory physiological parameters than reported in previous models. The benefit (achieving ischaemia) versus risk (lethal arrhythmia) of the duration of the coronary occlusion should be considered.

Grazing management and supplementation effects on forage and dairy cow performance on cool-season pastures in the southeastern United States.

Cool-season annual forages provide high-quality herbage for up to 5 mo in the US Gulf Coast states, but their management in pasture-based dairy systems has received little attention. Objectives of this study were to evaluate pasture and animal responses when lactating Holstein cows (n=32, mean DIM=184±21) grazed either N-fertilized rye (Secale cereale L.)-annual ryegrass (Lolium multiflorum Lam.) mixed pastures or rye-annual ryegrass-crimson clover (Trifolium incarnatum L.)-red clover (Trifolium pratense L.) pastures at 2 stocking rates (5 vs. 2.5 cows/ha) and 2 rates of concentrate supplementation [0.29 or 0.40 kg of supplement (as is)/kg of daily milk production]. Two cows paired by parity (one multiparous and one primiparous) were assigned randomly to each pasture. The 2 × 2 × 2 factorial arrangement of treatments was replicated twice in a completely randomized design. Forage mixture and supplementation rate did not affect milk production during three 28-d periods. Greater milk production occurred at the low (19.7 kg/d) than the high (14.7 kg/d) stocking rate during periods 2 and 3, but production was similar during period 1. Despite lower production per cow, milk production per hectare was generally greater at the high stocking rate (81.6 vs. 49.5 kg/ha). Generally, greater pregraze herbage mass on pastures at the lower stocking rate (1,400 vs. 1,150 kg/ha) accounted for greater herbage allowance. Both forage (8.0 vs. 5.9 kg/d) and total (14.1 vs. 11.6) organic matter intake were greater at the low stocking rate. Cows fed less supplement had greater forage organic matter intake (8.0 vs. 6.1 kg/d). Greater herbage mass was associated with the greater intake and subsequent greater milk production. Differences in forage nutritive value, blood metabolites and milk composition, although showing some response to treatments, may not be of sufficient magnitude to affect choice of pasture species or other management practices. Animal performance was not improved by adding clovers to mixed cool-season grass pastures like those in this study. Stocking rate had a major effect on pasture and animal performance. During the cool season, supplementation with concentrates should be planned based on estimated energy intake from forages to achieve optimum milk production and ensure maintenance of body condition.

Nitrate-nitrogen concentrations in the perched ground water under seepage-irrigated potato cropping systems.

Excessive nitrogen rates for potato production in northeast Florida have been declared as a potential source of nitrate pollution in the St. Johns River watershed. This 3-yr study examined the effect of N rates (0, 168, and 280 kg ha(-1)) split between planting and 40 d after planting on the NO(3)-N concentration in the perched ground water under potato (Solanum tuberosum cv. Atlantic) in rotation with sorghum sudan grass hybrid (Sorghum vulgare x Sorghum vulgare var. sudanese, cv. SX17), cowpea (Vigna unguiculata cv. Iron Clay), and greenbean (Phaseolus vulgare cv. Espada). Soil solution from the root zone and water from the perched ground water under potato were sampled periodically using lysimeters and wells, respectively. Fertilization at planting increased the NO(3)-N concentration in the perched ground water, but no effect of the legumes in rotation with potatoes on nitrate leaching was detected. Fertilization of green bean increased NO(3)-N concentration in the perched ground water under potato planted in the following season. The NO(3)-N concentration in the soil solution within the potato root zone followed a similar pattern to that of the perched ground water but with higher initial values. The NO(3)-N concentration in the perched ground water was proportional to the rainfall magnitude after potato planting. A significant increase in NO(3)-N concentration in the perched ground water under cowpea planted in summer after potato was detected for the side-dressing of 168 kg ha(-1) N applied to potato 40 d after planting but not at the 56 kg ha(-1) N side-dress. Elevation in NO(3)-N concentration in the perched ground water under sorghum was not significant, supporting its use as an effective N catch crop.

Changes in brown eggshell color as the hen ages.

The color of eggshells from eggs laid by commercial-type Hy-Line brown hens 25 wk of age was studied over a period of 10 mo. Color measurements were made by a color machine vision system and were analyzed using a mixed model to calculate between and within hen variances and to investigate the effect of time on shell color. Hens laid eggs with lighter colored shells as the flock aged, as evidenced by the lightness (L*) values increasing in time. A decrease in pigmentation was associated with a decrease in the amount of redness (a*) in the eggshell. When L* and a* values were corrected for egg weight, the rate of change in the L* and a* values decreased, indicating that size of the egg was a major factor affecting the color of the eggshell. These findings quantified the observations that older hens lay lighter colored eggs due to an increase in egg size associated with no proportionate change in the quantity of pigment deposited over the shell surface. Using a 2-stage sampling analysis and the variances between and within hens, sample sizes required to estimate the color of eggshells within 5% of the true mean were calculated. Accordingly, 11 eggs would need to be collected from each of the 51 hens housed for a study of brown eggshell color using the L*, a*, and b* (yellowness) coordinates.

Effects of exercise and oral antioxidant supplementation enriched in (n-3) fatty acids on blood oxidant markers and erythrocyte membrane fluidity in horses.

The aim of this study was to investigate in a placebo-controlled field study the effect of a (n-3)-vitamin supplementation on erythrocyte membrane fluidity (EMF), oxidant/antioxidant markers and plasmatic omega3/omega6 fatty acid ratio (FAR) in 12 eventing horses. Venous blood was sampled at rest before (PRE) and after (POST) a three week treatment period with either the supplement (group S, n=6) or a placebo (group P, n=6) as well as after 15min (POST E15') and 24h (POST E24h) after a standardised exercise test. The following markers were analysed: EMF, plasma antioxidant capacity of water and lipid soluble components, ascorbic acid, uric acid (UA), glutathione (reduced: GSH, oxidised: GSSG), vitamin E (Vit E), beta-carotene, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity, selenium, copper (Cu), zinc (Zn), oxidised proteins (Protox), lipid peroxides (Pool) and FAR. EMF did not differ between group S and P after treatment, but GPx remained unchanged in group S whereas it decreased in group P and plasma Cu/Zn ratio remained unchanged whereas it increased in group P. FAR were significantly increased in group S. Exercise induced a significant decrease of EMF (POST vs. E24h) in both groups, but which was significantly lower at E15' in group S than in group P. Exercise induced a significant increase of UA and ACW (POST vs. E15') and Protox (POST vs. E24h) in both groups. An exercise-related decrease in GSH and Pool (POST vs. E15') was found in group P, whereas Vit E and FAR (POST vs. E24h) significantly decreased in both groups. The study showed that exercise induced a decrease in EMF in horses associated with changes of blood oxidative balance. The (omega-3)-vitamin supplementation tested improved the oxidative balance poorly but delayed the exercise-induced decrease of EMF and increased the FAR.

Geographical variation in antibiotic resistance profiles of Escherichia coli isolated from swine, poultry, beef and dairy cattle farm water retention ponds in Florida.

AIMS: The aim of this study was to assess geographical variation in multiple antibiotic resistance (MAR) profiles of livestock Escherichia coli as well as to evaluate the ability of MAR profiles to differentiate sources of faecal pollution. METHODS AND RESULTS: More than 2000 E. coli isolates were collected from water retention ponds and manure of swine, poultry, beef and dairy farms in south, central and north Florida, and analysed for MAR using nine antibiotics. There were significant differences in antibiotic resistance of E. coli by season and livestock type for more than one antibiotic, but regional differences were significant only for ampicillin. Over the three regions, discriminant analysis using MAR profiles correctly classified 27% of swine, 49% of poultry, 56% of beef and 51% of dairy isolates. CONCLUSIONS: Regional variations in MAR combined with moderate discrimination success suggest that MAR profiles of E. coli may only be marginally successful in identifying sources of faecal pollution. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the existence of regional and seasonal differences in MAR profiles as well as the limited ability of MAR profiles to discriminate among livestock sources.

The effects of dietary N-3 and antioxidant supplementation on erythrocyte membrane fatty acid composition and fluidity in exercising horses.

REASONS FOR PERFORMING STUDY: Fatty acid supplementation could modulate erythrocyte membrane fluidity in horses at rest and during exercise, but information is lacking on the effect of exercise. OBJECTIVES: To assess the effect of exercise with, and without, an oral antioxidant supplementation enriched with n-3 fatty acids on erythrocyte membrane fluidity (EMF) and fatty acid composition in eventing horses. METHODS: Twelve healthy and regularly trained horses were divided randomly into 2 groups: group S received an oral antioxidant cocktail enriched in n-3 fatty acid (alphatocopherol, eicosapentaenoic acid [EPA] and docosahexaenoic acid [DHA]) whereas group P was placebo-treated. At the end of 4 weeks, all horses performed a standardised exercise test (ET) under field conditions. Venous blood was sampled before starting treatment (TO), immediately before (T1) as well as 15 min (T2) and 24 h (T3) after ET. Spin labelled (16-DOXYL-stearic acid) red blood cell membranes were characterised using the relaxation correlation time (Tc in inverse proportion to EMF). Fatty acid composition (%) of the membrane was determined by gas-liquid chromatography. RESULTS: Supplementation did not induce changes in EMF (T1 vs. TO) but significant changes in membrane composition were observed and there were increases in n-3 polyunsaturated fatty acid PUFA, n-3/n-6 ratio, and total n-3 fatty acids. Exercise (T2 vs. T1) induced a significant decrease of EMF in group P (Tc: +19%, P<0.05) and nonsignificant decrease in group S (Tc: +5%), whereas membrane fatty acid composition did not change in either group. During the recovery period (T3 vs. T2), EMF decreased significantly in group S (Tc: +29%, P<0.05) and nonsignificantly in group P (Tc: +18%) without any significant changes in fatty acid composition. CONCLUSION AND POTENTIAL RELEVANCE: An enriched oral antioxidant supplementation induced changes in membrane composition, which modulated the decrease in EMF induced by exercise. Long chain n-3 fatty acid supplementation might therefore be beneficial.

An environmental threshold for degree of phosphorus saturation in sandy soils.

There is critical need for a practical indicator to assess the potential for phosphorus (P) movement from a given site to surface waters, either via surface runoff or subsurface drainage. The degree of phosphorus saturation (DPS), which relates a measure of P already adsorbed by a soil to its P adsorption capacity, could be a good indicator of that soil's P release capability. Our primary objective was to find a suitable analytical protocol for determining DPS and to examine the possibility of defining a threshold DPS value for Florida's sandy soils. Four farmer-owned dairy sprayfields were selected within the Suwannee River basin and soil profiles were randomly obtained from each site, as well as from adjacent unimpacted sites. The soil samples were divided either by horizon or depth, and DPS was determined for each soil sample using ammonium-oxalate (DPS(Ox)), Mehlich-1 (DPS(M1)), and Mehlich-3 (DPS(M3)) extracts. All methods of DPS calculations were linearly related to one another (r2 > 0.94). Relationships between water-soluble P and DPS indicate that the respective change points are: DPS(Ox) = 20%, DPS(M1) = 20%, and DPS(M3) = 16%. These relationships include samples from Ap, E, and Bt horizons, and various combinations thereof, suggesting that DPS values can be used as predictors of P loss from a soil irrespective of the depth of the soil within a profile. Taking into consideration the change points, confidence intervals, agronomic soil test values, and DPS values from other studies, we suggest replacing Mehlich-1 P values in the Florida P Index with the three DPS categories (DPS(M1) = <30, 30-60, and >60%) to assign different P loss ratings in the P Index.

Comparison of three techniques for estimating the forage intake of lactating dairy cows on pasture.

Quantifying DMI is necessary for estimation of nutrient consumption by ruminants, but it is inherently difficult on grazed pastures and even more so when supplements are fed. Our objectives were to compare three methods of estimating forage DMI (inference from animal performance, evaluation from fecal output using a pulse-dose marker, and estimation from herbage disappearance methods) and to identify the most useful approach or combination of approaches for estimating pasture intake by lactating dairy cows. During three continuous 28-d periods in the winter season, Holstein cows (Bos taurus; n = 32) grazed a cool-season grass or a cool-season grass-clover mixture at two stocking rates (SR; 5 vs. 2.5 cows/ha) and were fed two rates of concentrate supplementation (CS; 1 kg of concentrate [as-fed] per 2.5 or 3.5 kg of milk produced). Animal response data used in computations for the animal performance method were obtained from the latter 14 d of each period. For the pulse-dose marker method, chromium-mordanted fiber was used. Pasture sampling to determine herbage disappearance was done weekly throughout the study. Forage DMI estimated by the animal performance method was different among periods (P < 0.001; 6.5, 6.4, and 9.6 kg/d for Periods 1, 2, and 3, respectively), between SR (P < 0.001; 8.7 [low SR] vs. 6.3 kg/d [high SR]) and between CS (P < 0.01; 8.4 [low CS] vs. 6.6 kg/d [high CS]). The period and SR effect seemed to be related to forage mass. The pulse-dose marker method generally provided greater estimates of forage DMI (as much as 11.0 kg/d more than the animal performance method) and was not correlated with the other methods. Estimates of forage DMI by the herbage disappearance method were correlated with the animal performance method. The difference between estimates from these two methods, ranging from -4.7 to 5.4 kg/d, were much lower than their difference from pulse-dose marker estimates. The results of this study suggest that, when appropriate for the research objectives, the animal performance or herbage disappearance methods may be useful and less costly alternatives to using the pulse-dose method.

Arsenic distribution in Florida urban soils: comparison between Gainesville and Miami.

Arsenic contamination is of concern due to its effect as a carcinogen. Understanding the distribution of arsenic in urban soils is important for establishing baseline concentrations from which anthropogenic effects can be measured. The soil cleanup target level (SCTL) for arsenic in Florida (0.8 and 3.7 mg kg(-1) in residential and commercial areas, respectively) is lower than in most states and is near the arsenic background concentrations in Florida soils. The objective of this study was to characterize the distribution of arsenic in the soils of two Florida cities, Gainesville and Miami. More than 200 soil samples were collected from three land-use classes in each city (residential, commercial, and public land), digested with USEPA Method 3051a, and analyzed with graphite furnace atomic absorption spectrophotometry. Arsenic concentrations varied greatly in Gainesville, ranging from 0.21 to approximately 660 mg kg(-1) with a geometric mean (GM) of 0.40 mg kg(-1) (after discarding outliers), which was significantly lower than the GM of 2.81 mg kg(-1) in Miami, although Miami samples ranged only from 0.32 to approximately 110 mg kg(-1). Arsenic concentrations in 29 and 4% of the Gainesville soil samples and 95 and 33% of the Miami samples exceeded the Florida residential and commercial SCTL, respectively. This study is the first to provide information on arsenic distribution in urban soils of Florida, and the data are useful for assessing arsenic contamination and determining the need for remediation.

Statistical issues in the analysis of low-dose endocrine disruptor data.

UNLABELLED: The National Institute of Environmental Health Sciences (NIEHS) and the U.S. Environmental Protection Agency (U.S. EPA) recently cosponsored the Endocrine Disruptors Low-Dose Peer REVIEW: The purpose of this meeting was to examine data supporting the presence or absence of low-dose effects of endocrine disruptors in specific studies and then to evaluate the likelihood and significance of these and/or other potential low-dose effects for humans. All invited speakers agreed to provide their raw data in advance of the meeting to a Statistics Subpanel, which was asked to reevaluate the authors' experimental design, data analysis, and interpretation of experimental results. The purpose of this statistical reevaluation was to provide an independent assessment of the experimental design and data analysis used in each of the studies and to identify key statistical issues relevant to the evaluation and interpretation of the data. This paper presents a summary of the Statistics Subpanel's evaluation. Specific examples are presented to illustrate problems that arose in the experimental design and data analysis of certain studies. The statistical principles and issues that are discussed in this paper are not unique to endocrine disruptor studies and should provide important guidelines regarding appropriate experimental design and statistical analysis for other types of laboratory investigations.

Discriminant analysis of ribotype profiles of Escherichia coli for differentiating human and nonhuman sources of fecal pollution.

Estuarine waters receive fecal pollution from a variety of sources, including humans and wildlife. Escherichia coli is a ubiquitous bacterium in the intestines of warm-blooded animals and is used as an indicator of fecal pollution. However, its presence does not specifically differentiate sources of pollution. A total of 238 E. coli isolates from human sources (HS) and nonhuman sources (NHS) were collected from the Apalachicola National Estuarine Research Reserve, from associated sewage treatment plants, and directly from animals and tested for ribotype (RT) profile. HS and NHS isolates showed 41 and 61 RT profiles, respectively. At a similarity index of ca. 50%, HS and NHS isolates demonstrated four clusters, with the majority of HS and NHS isolates located in clusters C and D; isolates obtained directly from human and animal feces also could be grouped within these clusters. Discriminant analysis (DA) of RT profiles showed that 97% of the NHS isolates and 100% of the animal fecal isolates were correctly classified. The average rate of correct classification for HS and NHS isolates was 82%. We conclude that DA of RT profiles may be a useful method for identifying HS and NHS fecal pollution and may potentially facilitate management practices.

Association of multiple-antibiotic-resistance profiles with point and nonpoint sources of Escherichia coli in Apalachicola Bay.

A total of 765 Escherichia coli isolates from point and nonpoint sources were collected from the Apalachicola National Estuarine Research Reserve, and their multiple-antibiotic-resistance (MAR) profiles were determined with 10 antibiotics. E. coli isolates from point sources showed significantly greater resistance (P < 0.05) to antibiotics and higher MAR indices than isolates from nonpoint sources. Specifically, 65 different resistance patterns were observed among point source isolates, compared to 32 among nonpoint source isolates. Examples of this contrast in MAR profiles included percentages of isolates with resistance to chlortetracycline-sulfathiazole of 33.7% and to chlortetracycline-penicillin G-sulfathiazole of 14.5% for point source isolates versus 15.4 and 1.7%, respectively, for nonpoint source isolates. MAR profile homology, based on coefficient similarity, showed that isolates from point sources were markedly more diverse than isolates from nonpoint sources. Seven clusters were observed among point source isolates, with a coefficient value of approximately 1.8. In contrast, only four clusters were observed among nonpoint source isolates. Covariance matrices of data displayed six very distinct foci representing nonpoint source E. coli isolates. Importantly, E. coli isolates obtained directly from human and animal feces also clustered among point and nonpoint sources, respectively. We conclude that E. coli MAR profiles were associated with point and nonpoint sources of pollution within Apalachicola Bay and that this method may be useful in facilitating management of other estuaries.