A Large, Refractory Nosocomial Outbreak of Klebsiella pneumoniae Carbapenemase-Producing Escherichia coli Demonstrates Carbapenemase Gene Outbreaks Involving Sink Sites Require Novel Approaches to Infection Control.

Carbapenem-resistant Enterobacteriaceae (CRE) represent a health threat, but effective control interventions remain unclear. Hospital wastewater sites are increasingly being highlighted as important potential reservoirs. We investigated a large Klebsiella pneumoniae carbapenemase (KPC)-producing Escherichia coli outbreak and wider CRE incidence trends in the Central Manchester University Hospital NHS Foundation Trust (CMFT) (United Kingdom) over 8 years, to determine the impact of infection prevention and control measures. Bacteriology and patient administration data (2009 to 2017) were linked, and a subset of CMFT or regional hospital KPC-producing E. coli isolates (n = 268) were sequenced. Control interventions followed international guidelines and included cohorting, rectal screening (n = 184,539 screens), environmental sampling, enhanced cleaning, and ward closure and plumbing replacement. Segmented regression of time trends for CRE detections was used to evaluate the impact of interventions on CRE incidence. Genomic analysis (n = 268 isolates) identified the spread of a KPC-producing E. coli outbreak clone (strain A, sequence type 216 [ST216]; n = 125) among patients and in the environment, particularly on 2 cardiac wards (wards 3 and 4), despite control measures. ST216 strain A had caused an antecedent outbreak and shared its KPC plasmids with other E. coli lineages and Enterobacteriaceae species. CRE acquisition incidence declined after closure of wards 3 and 4 and plumbing replacement, suggesting an environmental contribution. However, ward 3/ward 4 wastewater sites were rapidly recolonized with CRE and patient CRE acquisitions recurred, albeit at lower rates. Patient relocation and plumbing replacement were associated with control of a clonal KPC-producing E. coli outbreak; however, environmental contamination with CRE and patient CRE acquisitions recurred rapidly following this intervention. The large numbers of cases and the persistence of blaKPC in E. coli, including pathogenic lineages, are of concern.

Comparison of Nutrient and Metal Loadings with the Application of Swine Manure Slurries and Their Liquid Separates to Soils.

The accumulation of phosphorus (P) and metals is a serious concern with the continuous application of manure to agricultural soils. Solid-liquid separation of swine slurry is a promising approach to reduce P and metal loadings through application of separated liquid (SL) as a nutrient source. However, little information is available on nutrient and metal loadings with the application of SL compared with unseparated raw manure (RM). We analyzed element concentrations and calculated nutrient and metal loadings for RM and their respective SL applications, considering an application rate of 100 kg total nitrogen (N) ha. Samples of SL were obtained through three separation techniques: (i) centrifugation without a flocculant, (ii) centrifugation with a flocculant, and (iii) rotary press with a flocculant. Irrespective of separation technique, calculated P loadings with the application of SL were only 50 to 70% of that of RM at equivalent rates of total N yet exceeded crop removal rate. In contrast, calculated K and Na loadings with SL application were significantly greater than with RM, indicating a possible build-up of K and Na in soil. Calculated Ca and Mg loadings were significantly greater with RM than with SL. Loadings of Al, As, Ba, Cd, Cr, Fe, Mn, Ni, Pb, Sn, Se, Ti, and V were low, whereas Cu and Zn loadings were above crop removal rates for RM and SL. For solid-liquid separation to provide a lasting solution to the problem of P and metal accumulation, the SL must be supplemented with commercial N fertilizer to meet crop N demand.

Heavy-Metal Fractions in Solid and Liquid Separates of Swine Slurry Separated using Different Technologies.

Accumulation of metals is a concern with continuous application of swine slurry to agricultural soils. Solid-liquid separation is a promising approach for reducing phosphorus and total metal loadings with swine manure application to farmlands. However, very little work has been performed on the partitioning of different metal fractions in swine slurry to separated solids and liquids. This study examined the distribution of various metal fractions in raw manures (RM), their separated liquids (SL), and separated solids (SS). The three separation techniques used were centrifuge without flocculant (CNF), centrifuge with flocculant (CFL), and rotary press with flocculant (RFL). Concentrations of Cd, Cu, Zn, Ni, and Se in manure and separates were determined by a modified Sposito's sequential chemical fractionation scheme to extract water-soluble, exchangeable, organically bound, carbonate-precipitated, and residual fractions. The greatest concentrations of metals were recovered in the residual fraction, with the organically bound and carbonate-precipitated concentrations much greater than water-soluble and exchangeable fractions. Separation index () (i.e., percentage partitioned to SS) ranged from 13 to 66%, 9 to 87%, 16 to 93%, and 23 to 96% for water-soluble, exchangeable, organically bound, and carbonate-precipitated fractions, respectively. The values in general, were significantly ( < 0.05) greater for flocculant-based separation techniques than for CNF. For organically bound and carbonate-precipitated fractions, the greatest was obtained with the RFL for most metals. Our results suggest that applying the SL from RFL separation would minimize metal loading to farmlands compared with SL from CNF and CFL techniques. However, further validation is required using more sources of manure and different flocculants.

Phosphorus fractions in solid and liquid separates of Swine slurry separated using different technologies.

Solid-liquid separation is a manure management option whereby P-rich solid is separated from N-rich liquid, allowing the separated liquid to be used as a fertilizer without oversupplying P. Little information is available on how the different P fractions in manures are partitioned to solid and liquid during separation. We examined the distribution of various P fractions in liquid and solid separates of swine manure, separated using different techniques, to gain information useful for making choices regarding the optimum use of manure separates. Samples of raw manure (RM) and their separated solid (SS) and liquid (SL) were obtained using three different separation techniques: (i) centrifugation without flocculant (CNF), (ii) centrifugation with a flocculant (CFL), and (iii) rotary press with a flocculant (RFL). These were subsequently analyzed for P using a modified Hedley fractionation scheme. Only a small proportion of RM, ranging from 5 to 12%, was recovered in SS, an advantage if SS is to be transported off-site. Concentrations of molybdate-reactive P and total P in all P fractions were less in SL than in the corresponding RM on a fresh-weight basis. The separation index (percentage partitioned to SS) for total labile P (water-extractable + NaHCO-extractable P) was 63, 81, and 75% for CNF, CFL, and RFL, respectively. The proportion of total P in labile form was significantly lower in SL than in RM. Therefore, using SL as a fertilizer instead of RM may help to avoid excessive buildup of soil test P with manure applications.

Impact of manure phosphorus fractions on phosphorus loss from manured soils after incubation.

The risk of P loss from manured soils is more related to P fractions than total P concentration in manure. This study examined the impact of manure P fractions on P losses from liquid swine manure- (LSM), solid cattle manure- (SCM), and monoammonium phosphate- (MAP) treated soils. Manure or fertilizer was applied at 50 mg P kg soil, mixed, and incubated at 20°C for 6 wk to simulate the interaction between applied P and soil when P is applied well in advance of a high risk period for runoff. Phosphorus fractions in manure were determined using the modified Hedley fractionation scheme. We used simulated rainfall (75 mm h⁻¹ for 1 h) to quantify P losses in runoff from two soils (sand and clay loam). The proportion of total labile P (total P in water+NaHCO fractions) in manure was significantly greater in LSM (70%) than SCM (44%). Mean dissolved reactive P (DRP) load in runoff over 60 min was greatest from MAP-treated soil (18.1 mg tray⁻¹), followed by LSM- (14.0 mg tray⁻¹) and SCM- (11.0 mg tray⁻¹) treated soils, all of which were greater than mean DRP load from the check (5.2 mg tray⁻¹). Total labile P (water+NaHCO) in manure was a more accurate predictor of runoff DRP loads than water extractable P, alone, for these two soils. Therefore, NaHCO extraction of manure P may be a useful tool for managing the risk of manure P runoff losses when manure is applied outside a high risk period for runoff loss.

The seasonality of nitrate and phosphorus leaching from manure and chemical fertilizer added to a chernozemic soil in Canada.

Identifying seasons sensitive to nutrient losses could help farmers and policymakers to formulate effective nutrient loss reduction strategies. This long-term study monitored water percolation as well as nitrate (NO3 -N) and total phosphorus (TP) leaching from liquid swine manure and chemical fertilizer applied to intact core lysimeters in a sandy loam soil in Manitoba, Canada. Water percolation, NO3 -N, and TP leaching were monitored from 2005 to 2016. Chemical fertilizer showed greater average annual mean water percolation (p = .01), annual flow-weighted mean concentration (FWMC) of NO3 -N (22 mg L-1 ; p < .001), and annual NO3 -N leaching (36 kg N ha-1 ; p = .002) compared with the manure treatment (FWMC NO3 -N, 15 mg L-1 ; NO3 -N leaching load, 22 kg N ha-1 ). Average annual mean TP loss did not differ between treatments (p = .86). Spring (April-June) was the most sensitive season, when >75% of annual percolation, >80% of annual NO3 -N, and >68% of annual TP leaching losses occurred from both manure and chemical fertilizer. Annual NO3 -N and TP leaching increased exponentially with cumulative winter and spring precipitation (control, r2  = .69; manure, r2  = .79; chemical fertilizer, r2  = .63) and decreased with winter and spring air temperatures. The largest spring NO3 -N and TP leaching losses were observed in 2013, which followed the dry year of 2012, indicating the potential for nutrient flushing. The findings emphasize the need for environmentally sound N and P management strategies in cold North American regions underlain by coarse-textured soils, particularly during the spring season.

Phosphorus characteristics of Canada-wide animal manures and implications for sustainable manure management with a cleaner environment.

Animal manure can be a serious environmental concern if improperly managed, particularly with regard to phosphorus (P). A good understanding on manure P chemistry is required for improving manure management in an environmentally sustainable manner. In the study, 102 representative manure samples collected from farms of major intensive livestock areas of Canada were sequentially fractionated with H2O, 0.5 M NaHCO3, 0.1 M NaOH, and 1.0 M HCl, respectively, for measuring inorganic (Pi) and organic P (Po). Across the manures, total P (TP) ranged from 3.71 to 17.3 g kg-1, with total Pi and available P (i.e., the sum of H2O-Pi and NaHCO3-Pi) accounting for 67.0-92.4 % and 35.6-67.3 % of TP, respectively. Composting reduced the percentages of available P and Po in TP, and meanwhile increased the percentages of moderately stable HCl-Pi. Compared to other P fractions, manure available P increased more rapidly with increases in TP; once manure TP reached 7.8-9.6 g kg-1, further TP increase enhanced transformation to more recalcitrant P (i.e., NaOH-Pi and HCl-Pi). Under Canadian conditions, manure application to meet the demand of crops for N would lead to P buildup in the soil at rates of 6.1-41.6 kg P ha-1 yr-1, increasing runoff P loss risk. Manure compost and poultry manure tend to cause rapid P buildup in the soil after land application and become a long-term P source for runoff losses. The results help to develop scientifically-sound manure-specific P management technologies which would enable farmers to achieve sustainable crop production with improved environment.

Phosphorus fractions in manure from growing pigs receiving diets containing micronized peas and supplemental enzymes.

Different livestock feeds manipulations have been reported to reduce the total P concentration in manure. Information on the influence of these dietary manipulation strategies on the forms of P in manure is, however, limited. This study was, therefore, conducted to investigate the effect of diet manipulation through feed micronization and enzyme supplementation on the forms of P in swine manure. Eight growing pigs were fed four diets: barley-raw pea (BRP), barley-micronized pea (BMP), barley-raw pea with enzyme (BRPE), and barley-micronized pea with enzyme (BMPE) in a 4 x 4 Latin square design. Because we are interested in the effect of enzyme cocktail and pea micronization on manure P, we did not reduce the non-phytate P with enzyme addition in this study. The fecal material and urine were collected and analyzed for total P. Fecal material was fractionated to determine the total P in H2O-, NaHCO3-, NaOH-, and HCl-extractable fractions. The total P in the residual fractions was also determined. About 98% of the total P excreted by the pigs was found in the fecal material. Inclusion of micronized pea in pig diet did not have any significant effect (p > 0.1) on either the total P or the different P fractions in the manure. The labile P (the sum of H2O-P and NaHCO3-P) was significantly reduced (p < 0.05) by the addition of enzyme to swine diets. Pigs fed the BRPE and BMPE had 14 and 18% lower labile P, respectively, compared with pigs fed the BRP. Enzyme addition to pig diets reduced not only the total P in manure, but also the labile P fraction, which is of great environmental concern. Thus, the potential of P loss to runoff and the subsequent eutrophication can be reduced by enzyme addition to pig diets.

Environmental index for estimating the risk of phosphorus loss in calcareous soils of Manitoba.

The degree of phosphorus saturation (DPS) has been used in evaluating the risk of P loss from soil to runoff. While techniques are available for calculating DPS for acid soils, no widely used technique exists for neutral to calcareous soils that are typical of the Northern Great Plains, including Manitoba (Canada) soils. This study aimed to develop techniques of calculating the DPS of neutral to alkaline soils. Four measures of soil labile P and ten indices of P sorption capacity were used to calculate the DPS of 115 Manitoba soils. The various DPS calculated were evaluated using water-extractable ((H2O)) P as an index of P susceptibility to runoff loss. The DPS obtained using Olsen-extractable ((Ols)) P and the Langmuir adsorption maximum (ES(max)) ranged from 0.5 to 31.9% while those obtained from P(Ols) and the single-point adsorption index (P(150)) ranged from 0.9 to 73.9%. Of all the DPS evaluated, those that included P(Ols) and Mehlich 3-extractable ((M3)) P as the numerator with either P(150) or ES(max) as the denominator were fairly well correlated with P(H2O) (r values ranged between 0.45 and 0.63). Along with ES(max) and P(150), a new method of calculating DPS was formulated as the ratio of P(Ols) or P(M3) to Ca(M3) or (Ca + Mg)(M3). We found that the ratio of ammonium oxalate-extractable ((ox)) P to (Al + Fe)(ox), which has been widely used to calculate DPS in acid soils, was not suitable for neutral to alkaline soils of Manitoba. In these neutral to alkaline soils, Ca(M3) or (Ca + Mg)(M3) were better indices of P sorption capacity while P(Ols) and P(M3) provided better estimates of labile soil P. The DPS calculated using Ca(M3) or (Ca + Mg)(M3) were well correlated with P(H2O); however, they were numerically smaller than those obtained from the Langmuir adsorption maximum. As such, a saturation coefficient (alpha) with a value of 0.2 was generated to improve the numerical values of the newly estimated DPS. This new approach can be used to estimate the DPS in neutral and calcareous soils without the need to generate a P adsorption maximum.

Laboratory characterization of phosphorus in fresh and oven-dried organic amendments.

This study was performed to determine the forms of P and to examine the influence of oven-drying on P forms in different organic amendments. Samples of biosolids, beef and dairy cattle manures, and hog manures from sow and nursery barns were used in this study. Both fresh and oven-dried amendments were analyzed for inorganic (Pi), organic (Po), and total phosphorus using a modified Hedley fractionation technique. Water extracted about 10% of total biosolids P and 30 to 40% of total hog and cattle manure P. The amount of P extracted by NaHCO3 ranged from 21 to 32% of total P in all organic amendments except in the dairy cattle manure with 45% of total P. The labile P fraction (sum of H2O- and NaHCO3-extractable P) was 24% of biosolids P, 60% of hog manure P, and 70% of dairy cattle manure P. The residual P was about 10% in biosolids and cattle manures and 5 to 8% in hog manures. Oven-drying caused a transformation in forms of P in the organic amendments. In hog manures, H2O-extractable Po was transformed to Pi, while in the dairy manure NaHCO3-extractable P was converted to H2O-extractable Pi with oven-drying. Therefore, caution should be exercised in using oven-drying for studies that evaluate forms of P in organic amendments. Overall, these results indicate that biosolids P may be less susceptible to loss by water when added to agricultural land.