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Attempts to identify Clostridium botulinum toxin in milk from three experimentally intoxicated Holstein cows

Three adult lactating Holstein cows were injected in the subcutaneous abdominal vein with 175 ng/kg of body weight of Clostridium botulinum type C toxin (451 cow median toxic doses) to determine if this botulinum toxin crosses the blood–milk barrier. Whole blood (in sodium heparin) and clotted blood serum samples were taken at 0 min, 10 min, and 3, 6, 9, and 12 h postinoculation. Milk samples were taken at 0 min and at 3, 6, 9 and 12 h postinoculation. All samples were tested for the presence of the toxin using the mouse bioassay and immunostick ELISA test. The immunostick ELISA identified the toxin in whole blood and the mouse bioassay identified the toxin in serum at all times examined in all 3 animals. Toxin was not identified by either detection method in milk samples collected from the 3 animals. From these results, it appears that Clostridium botulinum type C toxin does not cross from the blood to the milk in detectable concentrations.

Journal of Dairy Science

Effect of diet on fecal and urinary estrogenic activity

The United States Environmental Protection Agency has identified estrogens from animal feeding operations as a major environmental concern, but few data are available to quantify the excretion of estrogenic compounds by dairy cattle. The objectives of this study were to quantify variation in estrogenic activity in feces and urine due to increased dietary inclusion of phytoestrogens. Ten Holstein heifers were assigned to 2 groups balanced for age and days pregnant; groups were randomly assigned to treatment sequence in a 2-period crossover design. Dietary treatments consisted of grass hay or red clover hay, and necessary supplements. Total collection allowed for sampling of feed refusals, feces, and urine during the last 4 d of each period. Feces and urine samples were pooled by heifer and period, and base extracts were analyzed for estrogenic activity (estrogen equivalents) using the yeast estrogen screen bioassay. Feces and urine samples collected from 5 heifers were extracted and analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to quantify excretion of 7 phytoestrogenic compounds. Excretion of 17-β estradiolequivalents in urine was higher and tended to be higher in feces for heifers fed red clover hay (84.4 and 120.2 mg/d for feces and urine, respectively) compared with those fed grasshay (57.4 and 35.6 mg/d). Analysis by LC-MS/MS indicated greater fecal excretion of equol, genistein, daidzein, coumestrol, and formononetin by heifers fed red clover hay (1634, 29.9, 96.3, 27.8, and 163 mg/d, respectively) than heifers fed grass hay (340, 3.0, 46.2, 8.8, and 18.3 mg/d, respectively). Diet had no effect on fecal biochanin A or 2-carbethoxy-5, 7-dihydroxy-4’-methoxyisoflavone. Four phytoestrogens were detected in urine (2-carbethoxy-5, 7-dihydroxy-4’-methoxyisoflavone, daidzein, equol, and formononetin) and their excretion was not affected by diet. Identifying sources of variation in estrogenic activity of manure will aid in the development of practices to reduce environmental estrogen accumulation.

Journal of Dairy Science

Environmental occurrence and shallow ground water detection of the antibiotic monensin from dairy farms

Pharmaceuticals used in animal feeding operations have been detected in various environmental settings. There is a growing concern about the impact on terrestrial and aquatic organisms and the development of antibiotic-resistant strains of microorganisms. Pharmaceutical use in milking cows is relatively limited compared with other livestock operations, except for the ionophore monensin, which is given to lactating cows as a feed. By weight, monensin can be the most significant antibiotic used in a dairy farm. This study investigates the potential of monensin to move from dairy operations into the surrounding ground water. Using two dairy farms in California as study sites, we twice collected samples along the environmental pathway-from flush lanes, lagoon waters, and shallow ground water beneath the dairies and beneath its associated manured fields. Monensin concentrations were determined using solid-phase extraction and liquid chromatography-tandem mass spectrometry with positive electrospray ionization. Monensin was detected in all of the flush lane and lagoon water samples. Theoretical maximum concentration estimated from the actual dosing rate and the theoretical excretion rate assuming no attenuation was one order of magnitude greater than observed concentrations, suggesting significant attenuation in the manure collection and storage system. Monensin was also detected, at levels ranging from 0.04 to 0.39 microg L(-1), in some of the ground water samples underneath the production area of the dairy but not from the adjacent manured fields. Concentrations in ground water immediately downgradient of the lagoons were one to two orders of magnitude lower than the concentrations detected in lagoons, suggesting attenuation in the subsurface. The data suggest the possibility of monensin transport into shallow (2-5 m) alluvial ground water from dairy management units, including manure storage lagoons and freestalls occupied by heifers, lactating cows, and dry cows.

Journal of Environmental Quality

Nitrogen losses from dairy manure estimated through nitrogen mass balance and chemical markers

Ammonia is an important air and water pollutant, but the spatial variation in its concentrations presents technical difficulties in accurate determination of ammonia emissions from animal feeding operations. The objectives of this study were to investigate the relationship between ammonia volatilization and ?? 15 N of dairy manure and the feasibility of estimating ammonia losses from a dairy facility using chemical markers. In Exp. 1, the N/P ratio in manure decreased by 30% in 14 d as cumulative ammonia losses increased exponentially. Delta 15 N of manure increased throughout the course of the experiment and ?? 15 N of emitted ammonia increased (p < 0.001) quadratically from -31??? to -15 ???. The relationship between cumulative ammonia losses and ?? 15 N of manure was highly significant (p < 0.001; r 2 = 0.76). In Exp. 2, using a mass balance approach, approximately half of the N excreted by dairy cows (Bos taurus) could not be accounted for in 24 h. Using N/P and N/K ratios in fresh and 24-h manure, an estimated 0.55 and 0.34 (respectively) of the N excreted with feces and urine could not be accounted for. This study demonstrated that chemical markers (P, K) can be successfully used to estimate ammonia losses from cattle manure. The relationship between manure ?? 15 N and cumulative ammonia loss may also be useful for estimating ammonia losses. Although promising, the latter approach needs to be further studied and verified in various experimental conditions and in the field. Copyright ?? 2009 by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America. All rights reserved.

Journal of Environmental Quality

Land management impacts on dairy-derived dissolved organic carbon in ground water

Dairy operations have the potential to elevate dissolved organic carbon (DOC) levels in ground water, where it may interact with organic and inorganic contaminants, fuel denitrification, and may present problems for drinking water treatment. Total and percent bioavailable DOC and total and carbon-specific trihalomethane (THM) formation potential (TTHMFP and STHMFP, respectively) were determined for shallow ground water samples from beneath a dairy farm in the San Joaquin Valley, California. Sixteen wells influenced by specific land management areas were sampled over 3 yr. Measured DOC concentrations were significantly elevated over the background as measured at an upgradient monitoring well, ranging from 13 to 55 mg L-1 in wells downgradient from wastewater ponds, 8 to 30 mg L-1 in corral wells, 5 to 12 mg L-1 in tile drains, and 4 to 15 mg L-1 in wells associated with manured fields. These DOC concentrations were at the upper range or greatly exceeded concentrations in most surface water bodies used as drinking water sources in California. DOC concentrations in individual wells varied by up to a factor of two over the duration of this study, indicating a dynamic system of sources and degradation. DOC bioavailability over 21 d ranged from 3 to 10%, comparable to surface water systems and demonstrating the potential for dairy-derived DOC to influence dissolved oxygen concentrations (nearly all wells were hypoxic to anoxic) and denitrification. TTHMFP measurements across all management units ranged from 141 to 1731 ??g L-1, well in excess of the maximum contaminant level of 80 ??g L-1 established by the Environmental Protection Agency. STHMFP measurements demonstrated over twofold variation (???4 to ???8 mmol total THM/mol DOC) across the management areas, indicating the dependence of reactivity on DOC composition. The results indicate that land management strongly controls the quantity and quality of DOC to reach shallow ground water and hence should be considered when managing ground water resources and in any efforts to mitigate contamination of ground water with carbon-based contaminants, such as pesticides and pharmaceuticals. Copyright ?? 2008 by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America. All rights reserved.

Journal of Environmental Quality

Sources and transformations of nitrate from streams draining varying land uses: Evidence from dual isotope analysis

Knowledge of key sources and biogeochemical processes that affect the transport of nitrate (NO 3 - ) in streams can inform watershed management strategies for controlling downstream eutrophication. We applied dual isotope analysis of NO 3 - to determine the dominant sources and processes that affect NO 3 - concentrations in six stream/river watersheds of different land uses. Samples were collected monthly at a range of flow conditions for 15 mo during 2004-05 and analyzed for NO 3 - concentrations, ?? 15 N NO3 , and ?? 18 O NO3 . Samples from two forested watersheds indicated that NO 3 - derived from nitrification was dominant at baseflow. A watershed dominated by suburban land use had three ?? 18 O NO3 values greater than +25???, indicating a large direct contribution of atmospheric NO 3 - transported to the stream during some high flows. Two watersheds with large proportions of agricultural land use had many ?? 15 N NO3 values greater than +9???, suggesting an animal waste source consistent with regional dairy farming practices. These data showed a linear seasonal pattern with a ?? 18 O NO3 :?? 15 N NO3 of 1:2, consistent with seasonally varying denitrification that peaked in late summer to early fall with the warmest temperatures and lowest annual streamflow. The large range of ?? 15 N NO3 values (10???) indicates that NO 3 - supply was likely not limiting the rate of denitrification, consistent with ground water and/or in-stream denitrification. Mixing of two or more distinct sources may have affected the seasonal isotope patterns observed in these two agricultural streams. In a mixed land use watershed of large drainage area, none of the source and process patterns observed in the small streams were evident. These results emphasize that observations at watersheds of a few to a few hundred km 2 may be necessary to adequately quantify the relative roles of various NO 3 - transport and process patterns that contribute to streamflow in large basins. Copyright ?? 2009 by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America. All rights reserved.

Journal of Environmental Quality