Jeffrey L. Ullman

pH and temperature effects on the hydrolysis of three β-lactam antibiotics: ampicillin, cefalotin and cefoxitin.

An understanding of antibiotic hydrolysis rates is important for predicting their environmental persistence. Hydrolysis rates and Arrhenius constants were determined as a function of pH and temperature for three common β-lactam antibiotics, ampicillin, cefalotin, and cefoxitin. Antibiotic hydrolysis rates at pH4-9 at 25 °C, 50 °C, and 60 °C were quantified, and degradation products were identified. The three antibiotics hydrolyzed under ambient conditions (pH7 and 25 °C); half-lives ranged from 5.3 to 27 d. Base-catalyzed hydrolysis rates were significantly greater than acid-catalyzed and neutral pH hydrolysis rates. Hydrolysis rates increased 2.5- to 3.9-fold for a 10 °C increase in temperature. Based on the degradation product masses found, the likely functional groups that underwent hydrolysis were lactam, ester, carbamate, and amide moieties. Many of the proposed products resulting from the hydrolysis of ampicillin, cefalotin, and cefoxitin likely have reduced antimicrobial activity because many products contained a hydrated lactam ring. The results of this research demonstrate that β-lactam antibiotics hydrolyze under ambient pH and temperature conditions. Degradation of β-lactam antibiotics will likely occur over several weeks in most surface waters and over several days in more alkaline systems.

The effects of the antibiotics ampicillin, florfenicol, sulfamethazine, and tylosin on biogas production and their degradation efficiency during anaerobic digestion.

The impacts of four common animal husbandry antibiotics (ampicillin, florfenicol, sulfamethazine, and tylosin) on anaerobic digestion (AD) treatment efficiency and the potential for antibiotic degradation during digestion were evaluated. Sulfamethazine and ampicillin exhibited no impact on total biogas production up to 280 and 350 mg/L, respectively, although ampicillin inhibited biogas production rates during early stages of AD. Tylosin reduced biogas production by 10-38% between 130 and 913 mg/L. Florfenicol reduced biogas by ≈ 5%, 40% and 75% at 6.4, 36 and 210 mg/L, respectively. These antibiotic concentrations are higher than commonly seen for mixed feedlot manure, so impacts on full scale AD should be minimal. Antibiotic degradation products were found, confirming AD effectively degraded ampicillin, florfenicol, and tylosin, although some products were persistent throughout the process. Contamination of AD solid and liquid effluents with sulfamethazine and antibiotic transformation products from florfenicol and tylosin could present an environmental concern.

Urine from Treated Cattle Drives Selection for Cephalosporin Resistant Escherichia coli in Soil

The U.S. Food and Drug Administration recently issued new rules for using ceftiofur in food animals in part because of an increasing prevalence of enteric bacteria that are resistant to 3rd-generation cephalosporins. Parenteral ceftiofur treatment, however, has limited effects on enteric bacteria so we tested the hypothesis that excreted ceftiofur metabolites exert significant selection pressure for ceftiofur-resistant Escherichia coli in soil. Test matrices were prepared by mixing soil with bovine feces and adding urine containing ceftiofur metabolites (CFM) (0 ppm, ∼50 ppm and ∼100 ppm). Matrices were incubated at 23°C or 4°C for variable periods of time after which residual CFM was quantified using a bioassay. Bla CMY-2 plasmid-bearing ceftiofur resistant (cefR) E. coli and one-month old calves were used to study the selection effects of CFM and transmission of cefR bacteria from the environment back to animals. Our studies showed that urinary CFM (∼13 ppm final concentration) is biologically degraded in soil within 2.7 days at 23°C, but persists up to 23.3 days at 4°C. Even short-term persistence in soil provides a >1 log10 advantage to resistant E. coli populations, resulting in significantly prolonged persistence of these bacteria in the soil (∼two months). We further show that resistant strains readily colonize calves by contact with contaminated bedding and without antibiotic selection pressure. Ceftiofur metabolites in urine amplify resistant E. coli populations and, if applicable to field conditions, this effect is far more compelling than reported selection in vivo after parenteral administration of ceftiofur. Because ceftiofur degradation is temperature dependent, these compounds may accumulate during colder months and this could further enhance selection as seasonal temperatures increase. If cost-effective engineered solutions can be developed to limit ex vivo selection, this may limit proliferation for ceftiofur resistant enteric bacteria while preserving the ability to use this important antibiotic in food animal production.

Hydrolysis of amphenicol and macrolide antibiotics: Chloramphenicol, florfenicol, spiramycin, and tylosin

Antibiotics that enter the environment can present human and ecological health risks. An understanding of antibiotic hydrolysis rates is important for predicting their environmental persistence as biologically active contaminants. In this study, hydrolysis rates and Arrhenius constants were determined as a function of pH and temperature for two amphenicol (chloramphenicol and florfenicol) and two macrolide (spiramycin and tylosin) antibiotics. Antibiotic hydrolysis rates in pH 4-9 buffer solutions at 25°C, 50°C, and 60°C were quantified, and degradation products were characterized. All of the antibiotics tested remained stable and exhibited no observable hydrolysis under ambient conditions typical of aquatic ecosystems. Acid- and base-catalyzed hydrolysis occurred at elevated temperatures (50-60°C), and hydrolysis rates increased considerably below pH 5 and above pH 8. Hydrolysis rates also increased approximately 1.5- to 2.9-fold for each 10°C increase in temperature. Based on the degradation product masses found, the functional groups that underwent hydrolysis were alkyl fluoride, amide, and cyclic ester (lactone) moieties; some of the resultant degradation products may remain bioactive, but to a lesser extent than the parent compounds. The results of this research demonstrate that amphenicol and macrolide antibiotics persist in aquatic systems under ambient temperature and pH conditions typical of natural waters. Thus, these antibiotics may present a risk in aquatic ecosystems depending on the concentration present.

Analysis of wheat prolamins, the causative agents of celiac sprue, using reversed phase high performance liquid chromatography (RP-HPLC) and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS).

Wheat prolamins, commonly known as “gluten”, are a complex mixture of 71–78 proteins, which constitute ~80% of the proteins in the wheat grains and supply 50% of the global dietary protein demand. Prolamins are also responsible for numerous gluten-induced disorders and determine the unique visco-elastic properties of the wheat dough. These properties necessitate the reliable determination of the prolamin composition in wheat grains and their derived products. Therefore, this study examined the impact of HPLC conditions, including column type, column temperature, flow rate, and the gradient of polar and non-polar solvents in the mobile phase, to improve the analytical resolution of prolamins. The following conditions were found optimal for analyses: column temperature 60 °C, flow rate 1.0 mL/min and an elution gradient of 20%–60% of 0.1% trifluoroacetic acid + acetonitrile in 60 min. For further improvement of resolution, gliadin and glutenin extracts were analyzed using MALDI-TOF-MS in combination with HPLC fractionation. Two semi-quantitative methods, densitometry of stained polyacrylamide gels and HPLC, were used to determine relative prolamin quantities and the correspondence between the methods was established. The combinatorial gluten analyses approach developed during the present study was used to analyze prolamin profiles of wheat transformants expressing DEMETER silencing artificial microRNA, and the results are discussed.

Intensified nitrogen removal of constructed wetland by novel integration of high rate algal pond biotechnology.

High rate algal pond (HRAP) was combined with constructed wetland (CW) to intensify nitrogen removal through optimizing nitrification and denitrification. Nitrification and denitrification process mainly depends on the oxygen content and carbon source level in CWs. Algal biomass was enriched in HRAP, and dissolved oxygen (DO) concentration was increased via photosynthesis. Algal debris increased COD as degradable bioresource. The results showed that HRAP-CW hybrid systems effectively promoted the nitrogen removal performance due to rich DO and COD. The extension of hydraulic retention time in HRAP significantly improved NH4-N and TN removals by 10.9% and 11.1% in hybrid systems, respectively. The highest NH4-N and TN removals in hybrid systems respectively reached 67.2% and 63.5%, which were significantly higher than those in single CW. The study suggested that the hybrid system had the application potentials in nitrogen removal from wastewater.

Sediment and Nutrient Reduction in Irrigation Return Flows by Vegetated Filter Strips on Surface Irrigated Fields

Nonpoint sources account for nearly two-thirds of nutrient loading to the nations surface waters (Beutel et al., 2009; Sharpley et al., 2001). Vegetated filter strips can mitigate the effects of agricultural activities by presenting a physical barrier to sediment, nutrients, and pesticides being carried into streams (Munoz-Carpena and Parsons, 2004; Krutz et al., 2005). Filter strips may also reduce the flux of soluble nutrients through plant uptake or by supporting environmental conditions that favor chemical transformations such as denitrification (Haycock and Pinay, 1993; Cooper and Gilliam, 1987; Hickey and Doran, 2004).

The Development and Evaluation of a Portable Rainfall Simulator Capable of Mimicking Variable Rainfall Patterns

Rainfall simulators present a useful tool for investigating the impacts of land management practices on runoff and erosion processes associated with storm events. However, it is critical that these simulators generate accurate rainfall patterns that are representative of the area studied. Researchers have developed rainfall simulators specific for use in a particular region, but these systems largely lack the capability to mimic rainfall characteristics across differing climactic regimes. This inflexibility can be problematic when working in a region that exhibits considerable rainfall variability. For instance, the Pacific Northwest displays a wide range of precipitation attributes that vary from maritime to mountainous to inland climates.

Impacts of Chemical Fallow Practices in Conservation Tillage Management Systems on Runoff and Erosion in the Pacific Northwest

Annual precipitation rates of 350-400 mm (14-16 in) in the western portions of the Palouse region in eastern Washington limit dryland wheat production. Growers typically adopt fallow management practices as part of a regular rotation to increase available soil moisture for the subsequent crop. Traditionally, repeated tillage (tillage fallow) has been used to mechanically kill weeds to eliminate transpirational water loss. Conservation tillage in the region often entails the use of direct seed tillage systems coupled with herbicide use as an alternative weed control agent during the fallow period (chemical fallow or chem-fallow) to minimize soil disturbance. Although previous studies have examined chemical fallow implementation, most research has been conducted at research field stations. The objective of this study is to evaluate the potential conservation benefits of using chemical fallow on operational farms, providing insight into runoff and erosion control under “real world” scenarios.