Targeted 16S rRNA high-throughput sequencing to characterize microbial communities during composting of livestock mortalities.

AIM: A comprehensive understanding of the microbial community is necessary to ensure a significant reduction in pathogens during the composting process. METHODS AND RESULTS: Two biosecure, static composting systems containing cattle mortalities were constructed at subzero temperatures. Temperature at each sampling site was measured continuously and samples were grouped as either ≤50 or ≥55°C, based on temperature exposure required for effective pathogen inactivation during composting. High-throughput 454 sequencing was used to characterize the bacterial communities within each sample. Clustering of bacterial communities was observed according to temperature. However, neither richness nor diversity differed between temperature groups. Firmicutes was the most abundant phylum within both temperature groups but was more pronounced (63·6%) in samples ≥55°C (P < 0·05). Similarly, members of Clostridia, Clostridium sensu stricto (3·64%), Clostridium XI (0·59%), UF (Clostridiaceae 1) (5·29%) and UF (Clostridiales Incertae Sedis XI) (6·20%), were prominent at ≥55°C (P < 0·05), likely a reflection of spore survival and/or anaerobic microenvironments within passively aerated compost piles. Members of Thermobifida (3·54%), UO (Actinomycetales) (12·29%) and UO (Bacillales) (19·49%) were also prominent at ≥55°C (P < 0·05). CONCLUSION: Substantial spatial diversity exists within bacterial communities in field-scale compost piles. Localized temperature at the site of sampling may be one of the factors contributing to this phenomenon. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to describe the microbial community profile with the use of targeted 16S rRNA high-throughput sequencing in passively aerated composted livestock mortalities.

Effects of the chop lengths of alfalfa silage and oat silage on feed intake, milk production, feeding behavior, and rumen fermentation of dairy cows.

Effects of chop length (shorter: 6 mm, or longer: 19 mm) of alfalfa silage and oat silage were determined in 16 mid-lactation Holstein cows, 4 of which were rumen cannulated, using a replicated 4 x 4 Latin square design with a 2 x 2 arrangement of treatments. Experimental periods were 21 d long and consisted of 14 d of adaptation and 7 d of sampling. Cows received a total mixed ration containing [dry matter (DM) basis] 42.0% barley grain-based energy supplement, 10% protein supplement, and 24% of DM longer chop or shorter chop alfalfa silage and 24% of DM longer chop or shorter chop oat silage. Rumen pH was measured continuously, and rumen liquid flow rates were determined in rumen-cannulated cows. Feeding behavior was determined by videotaping, and meal patterns were determined by continuously weighing the feed in the bunk of 8 cows. Reducing the chop length of alfalfa silage and oat silage reduced the average geometric particle length from 14.2 to 10.9 mm and from 13.4 to 10.4 mm, respectively. Reducing the alfalfa chop length did not affect feed intake, whereas reducing the oat silage chop length increased DM intake from to 19.4 to 21.2 kg/d. Reducing the chop lengths of alfalfa silage and oat silage chop length did not affect milk production, rumen fermentation, feeding behavior, meal patterns, and blood metabolites. Daily milk yield, milk fat percentage, and milk protein percentage averaged 36.1 kg/d, 3.00%, and 3.16%, respectively, across diets. The low milk fat percentages suggest that the diets induced subacute ruminal acidosis. This was also substantiated by the rumen pH, which was below 5.6 for more than 122 min/d for all diets. The onset of subacute ruminal acidosis despite apparently adequate dietary neutral detergent fiber content and particle size distribution as well as the long duration of chewing might be attributed to sorting against long feed particles.

Effects of chop length of alfalfa and corn silage on milk production and rumen fermentation of dairy cows.

Effects of chop length (shorter = 10 mm or longer = 19 mm) of alfalfa silage and corn silage were determined in 16 midlactation Holstein cows using a 4 x 4 Latin square design with a 2 x 2 arrangement of treatments. Experimental periods were 21 d long and consisted of 14 d of adaptation and 7 d of sampling. Cows received total mixed ration containing (dry matter basis) 44.0% barley grain-based energy supplement, 12.6% protein supplement, and 21.7% longer chop or shorter chop alfalfa silage and 21.7% longer chop or shorter chop corn silage. Reducing the chop length of alfalfa silage and corn silage reduced the average geometric particle length from 14.4 to 11.0 mm and from 14.2 to 10.4 mm, respectively. Reducing the chop length of both silages reduced the proportion of the diets retained by the 8- and 19-mm screen of the Penn State Particle Separator from 55.0 to 46.0% of dry matter. Reducing the alfalfa chop length increased total rumen volatile fatty acids at 4 to 5 h after feeding but did not affect rumen pH at 4 to 5 h after feeding, feed intake, and milk production. Reducing the corn silage chop length increased dry matter intake from 22.3 to 23.2 kg/d, increased rumen pH at 4 to 5 h after feeding from 6.12 to 6.20, but did not alter rumen volatile fatty acids at 4 to 5 h after feeding or milk production. Daily milk yield, milk fat percentage, and milk protein percentage averaged 38.2 kg/d, 2.62%, and 3.29%, respectively, across all diets. The low milk fat percentages suggest that all diets induced subacute ruminal acidosis (SARA), whereas the rumen pH did not indicate SARA. This discrepancy could be due to a difference in the time of rumen pH measurement and the time of the lowest rumen pH. Hence, the pH data need to be interpreted with caution. Diets could have induced SARA, because for all experimental diets the content of forage neutral detergent fiber was lower than recommended for barley grain-based diets.

BOARD-INVITED REVIEW: Quantifying water use in ruminant production.

The depletion of water resources, in terms of both quantity and quality, has become a major concern both locally and globally. Ruminants, in particular, are under increased public scrutiny due to their relatively high water use per unit of meat or milk produced. Estimating the water footprint of livestock production is a relatively new field of research for which methods are still evolving. This review describes the approaches used to quantify water use in ruminant production systems as well as the methodological and conceptual issues associated with each approach. Water use estimates for the main products from ruminant production systems are also presented, along with possible management strategies to reduce water use. In the past, quantifying water withdrawal in ruminant production focused on the water demand for drinking or operational purposes. Recently, the recognition of water as a scarce resource has led to the development of several methodologies including water footprint assessment, life cycle assessment, and livestock water productivity to assess water use and its environmental impacts. These methods differ with respect to their target outcome (efficiency or environmental impacts), geographic focus (local or global), description of water sources (green, blue, and gray), handling of water quality concerns, the interpretation of environmental impacts, and the metric by which results are communicated (volumetric units or impact equivalents). Ruminant production is a complex activity where animals are often reared at different sites using a range of resources over their lifetime. Additional water use occurs during slaughter, product processing, and packaging. Estimating water use at the various stages of meat and milk production and communicating those estimates will help producers and other stakeholders identify hotspots and implement strategies to improve water use efficiency. Improvements in ruminant productivity (i.e., BW and milk production) and reproductive efficiency can also reduce the water footprint per unit product. However, given that feed production makes up the majority of water use by ruminants, research and development efforts should focus on this area. More research and clarity are needed to examine the validity of assumptions and possible trade-offs between ruminants' water use and other sustainability indicators.

Application of fecal near-infrared reflectance spectroscopy profiling for the prediction of diet nutritional characteristics and voluntary intake in beef cattle.

The objective of this study was to evaluate the use of fecal near-infrared reflectance spectroscopy (NIRS) profiling to predict diet nutritional characteristics and voluntary DMI in beef cattle. Fecal samples were collected for growing cattle across 11 experiments in which individual animal performance and DMI was measured. Dried and ground fecal composite samples collected from each animal were subjected to fecal NIRS analysis by a Foss NIRS 6500 scanning monochromator (Foss, Eden Prairie, MN) at the Grazingland Animal Nutrition Laboratory (Temple, TX). Fecal spectra were then used to develop equations to predict diet composition (trials 1 to 11; = 408), digestibility (trials 1 to 5; = 155), and DMI (trials 1 to 11; = 408). Coefficients of determination for calibration () and cross-validation () for prediction of diet nutritional characteristics were lower for NDF ( = 0.85; = 0.82) than for CP ( = 0.90; = 0.88). For the prediction of DMI, and ranged from 0.69 and 0.67 for the prediction of trial-average DMI to 0.76 and 0.73 for the prediction of fecal-collection-period DMI. While the and obtained for the prediction of DMI were lower than those obtained for the prediction of diet composition or digestibility, fecal NIRS prediction equations for DMI were successful in predicting the mean DMI of groups, as no differences were found for the prediction of fecal-collection-period DMI (Diff. = 1.10; = 0.72) or trial DMI (Diff. = -0.47; = 0.86).

The relationship between scrotal circumference at weaning and at one year of age in beef bulls.

Scrotal circumference data obtained from performance test station bulls were corrected to 240 and 365 days of age. Data were analyzed, retrospectively, to determine the outcome for scrotal circumference at 365 days of age, if bulls were culled at selected cut-off values for scrotal circumference at 240 days of age. The data indicate that scrotal circumference measurements in weaned bulls may not be useful as a culling tool, since a large proportion of bulls, irrespective of breed, fell below the selected cut-off measurement for scrotal circumference at weaning, but met the minimum scrotal circumference at one year of age. Scrotal circumference measurements at 240 days of age may, however, be useful as a tool for selecting bulls with a greater than 80% probability of achieving the minimum scrotal circumference measurement at one year of age.

Reducing GHG emissions through genetic improvement for feed efficiency: effects on economically important traits and enteric methane production.

Genetic selection for residual feed intake (RFI) is an indirect approach for reducing enteric methane (CH4) emissions in beef and dairy cattle. RFI is moderately heritable (0.26 to 0.43), moderately repeatable across diets (0.33 to 0.67) and independent of body size and production, and when adjusted for off-test ultrasound backfat thickness (RFI fat) is also independent of body fatness in growing animals. It is highly dependent on accurate measurement of individual animal feed intake. Within-animal repeatability of feed intake is moderate (0.29 to 0.49) with distinctive diurnal patterns associated with cattle type, diet and genotype, necessitating the recording of feed intake for at least 35 days. In addition, direct measurement of enteric CH4 production will likely be more variable and expensive than measuring feed intake and if conducted should be expressed as CH4 production (g/animal per day) adjusted for body size, growth, body composition and dry matter intake (DMI) or as residual CH4 production. A further disadvantage of a direct CH4 phenotype is that the relationships of enteric CH4 production on other economically important traits are largely unknown. Selection for low RFI fat (efficient, -RFI fat) will result in cattle that consume less dry matter (DMI) and have an improved feed conversion ratio (FCR) compared with high RFI fat cattle (inefficient; +RFI fat). Few antagonistic effects have been reported for the relationships of RFI fat on carcass and meat quality, fertility, cow lifetime productivity and adaptability to stress or extensive grazing conditions. Low RFI fat cattle also produce 15% to 25% less enteric CH4 than +RFI fat cattle, since DMI is positively related to enteric methane (CH4) production. In addition, lower DMI and feeding duration and frequency, and a different rumen bacterial profile that improves rumen fermentation in -RFI fat cattle may favor a 1% to 2% improvement in dry matter and CP digestibility compared with +RFI fat cattle. Rate of genetic change using this approach is expected to improve feed efficiency and reduce enteric CH4 emissions from cattle by 0.75% to 1.0% per year at equal levels of body size, growth and body fatness compared with cattle not selected for RFI fat.

Screening of condensed tannins from Canadian prairie forages for anti-Escherichia coli O157:H7 with an emphasis on purple prairie clover (Dalea purpurea Vent).

Tannins from forages grown (n = 10) on the Canadian prairie, as well as from Quebracho, Rhus semialata, and brown seaweed (Ascophyllum nodosum), were screened for anti-Escherichia coli O157:H7 activity against E. coli O157:H7 strain 3081 at a concentration of 400 µg/ml for each tannin type, except for brown seaweed, which was at 50 µg/ml. Growth of the bacteria was assessed by measuring the optical density at 600 nm over 24 h. Tannin from seaweed at a concentration of 50 µg/ml inhibited growth of strain 3081. Among the terrestrial forages, only condensed tannins (CT) from purple prairie clover (Dalea purpurea Vent; PPC) increased (P < 0.05) the lag time and reduced (P < 0.05) the growth rate of E. coli O157:H7. The anti-E. coli O157:H7 activity of PPC CT was further assessed by culturing E. coli strain ATCC 25922 and eight strains of E. coli O157:H7 with PPC CT at 0, 25, 50, 100, or 200 µg/ml. Selected strains were enumerated after 0, 6, and 24 h of incubation, and fatty acid composition was determined after 24 h of incubation. E. coli strain 25922 was cultured with 0, 50, or 200 µg of CT per ml and harvested during the exponential growth phase for examination by transmission electron microscopy. Increasing CT concentration linearly increased (P < 0.001) the lag times of seven strains and linearly reduced (P < 0.001) the growth rates of eight E. coli O157:H7 strains. Proportions of unsaturated fatty acids in the total fatty acids were decreased (P < 0.01) by CT at 50 µg/ml. Transmission electron microscopy showed that CT disrupted the outer membrane structure. Anti-E. coli O157:H7 activity of PPC CT at levels of up to 200 µg/ml was bacteriostatic rather than bactericidal, and the mechanism of anti-E. coli activity may involve alteration in the fatty acid composition and disruption of the outer membrane of the cell.

Efficacy of ionophores in cattle diets for mitigation of enteric methane.

Use of ionophores in cattle diets has been proposed as a strategy for mitigation of enteric CH4 emissions. Short- and long-term effects of feeding a single ionophore (monensin) or rotation of 2 ionophores (monensin and lasalocid) on enteric CH4 emissions were evaluated in 36 Angus yearling steers (328 +/- 24.9 kg of BW) over a 16-wk period. Steers were randomly assigned to 6 dietary treatments of 6 steers each. The 6 diets were low-concentrate without ionophore supplementation, low-concentrate with monensin supplementation, low-concentrate with a 2-wk rotation of monensin and lasalocid supplementation, high-concentrate without ionophore supplementation, high-concentrate with monensin supplementation, and high-concentrate with a 2-wk rotation of monensin and lasalocid supplementation. Daily enteric CH4 emissions, as measured using the SF(6) tracer gas technique, ranged from 54.7 to 369.3 L/steer daily. Supplementing ionophores decreased (P < 0.05) enteric CH4 emissions, expressed as liters per kilogram of DMI or percentage of GE intake, by 30% for the first 2 wk and by 27% for the first 4 wk, for cattle receiving the high-concentrate and low-concentrate diets, respectively. Cattle fed a rotation of ionophores did not (P > 0.05) exhibit a greater decrease and did not (P > 0.05) have a longer period of depressed enteric CH4 emissions compared with cattle receiving monensin only. Ionophore supplementation did not (P > 0.05) alter total ruminal fluid VFA concentration; however, the acetate:propionate ratio and ammonia-N concentration in ruminal fluid were decreased (P < 0.001) from the time that ionophores were introduced to the time they were removed from the diets. Both monensin and the rotation of monensin and lasalocid decreased (P < 0.001) total ciliate protozoal populations by 82.5% in the first 2 wk and by 76.8% in the first 4 wk during which they were supplemented in the high-concentrate and low-concentrate diets, respectively. Original ciliate protozoal populations were restored by the fourth and sixth week of supplementation when cattle were fed the high- or low-concentrate diets, respectively. No significant change was observed thereafter. These data suggest that the effects of ionophores on enteric CH(4) production are related to ciliate protozoal populations and that ciliate protozoal populations can adapt to the ionophores present in either low- or high-concentrate diets. Rotation of monensin and lasalocid did not (P > 0.05) prevent ciliate protozoal adaptation to ionophores.

Ochratoxin A as a contaminant in the human food chain: a Canadian perspective.

Penetration of ochratoxin A into the human food chain in Canada was assessed by analysing stored grains, porcine blood and, finally, human blood. The potential for mycological growth and production of ochratoxin A was determined in 164 samples of stored grain (barley, wheat, maize and silage) collected from producers in Manitoba. A total of 34% were found to have the capacity to produce ochratoxin A; 14.5% had concentrations greater than 1.0 mg/kg. In 1988, 1200 blood samples were obtained from swine destined for slaughter in western Canada. High-performance liquid chromatography demonstrated that 3.6% of the 194 blood samples collected in February and March and 4.2% of the 1006 collected in May, June and July had concentrations of ochratoxin A that exceeded 20 ng/ml. In a subsequent survey of porcine blood carried out in 1989-90, 16-65% of the samples had detectable levels of ochratoxin A, at mean concentrations of 5.4-19.4 ng/ml. Subsequently, human blood samples were collected from 159 individuals, 69 of whom had some form of renal impairment. Of the latter, 40% had detectable levels of ochratoxin A, and 12% had concentrations greater than 0.5 ng/ml. Of the non-renal patients, 39% had detectable levels of ochratoxin A, and 11% had concentrations greater than 0.5 ng/ml. These studies demonstrate that ochratoxin A is present in the blood of people in Canada and that two possible points of entry of this toxin into the human food chain are contaminated grain and pork products.

Physiological and production responses to feeding schedule in lactating dairy cows exposed to short-term, moderate heat stress.

The objective of this research was to characterize the production responses of lactating dairy cows during and after short-term, moderate heat exposure, and to determine whether evening (p.m.) feeding would alleviate the associated production losses. In a two-period, cross-over design, eight mature lactating cows were fed a total mixed ration at either 0830 or 2030 h. Each 15-d period consisted of a 5-d thermoneutral phase, a 5-d heat stress phase and a 5-d thermoneutral recovery phase. Mean daily vaginal temperature and respiration rate increased by 0.6 +/- 0.04 degrees C and 27 +/- 1.3 breaths/min, respectively, during short-term heat exposure. Daily dry matter intake, milk yield and solids-not-fat were depressed by 1.4 +/- 0.13 kg, 1.7 +/- 0.32 kg and 0.07 +/- 0.023%, respectively, during heat exposure. During the recovery phase, dry matter intake remained depressed, milk protein declined by 0.05 +/- 0.020%, and daily milk yield exhibited a further decline of 1.2 +/- 0.32 kg. Time of feeding had no effect on vaginal temperature, respiration rate, dry matter intake, water intake, milk yield, fat-corrected milk, protein percent, solids-non-fat percent or somatic cell count during heat exposure or during the recovery period that followed. Fat percent was, however, significantly lower in p.m.-fed animals. These data indicate that short-term, moderate heat stress, which occurs during the spring and summer months in Canada and the Northern United States, will significantly decrease production in the lactating cow. Shifting from morning to evening feeding did not alleviate production losses associated with this type of heat stress.

The incidence and distribution of ochratoxin A in western Canadian swine.

A survey of swine destined for slaughter in Manitoba was conducted to examine the incidence of ochratoxin A (OA) in swine herds from different regions of Manitoba throughout the year 1989-90. Thirty-six percent of the serum samples which were collected from 1600 pigs contained detectable levels of OA. The identity of this toxin was confirmed using liquid chromatography-mass spectrometry and enzymatic hydrolysis. There was a significant effect of the region from which the herds originated, as well as the season in which the samples were collected on both the incidence (p < 0.001) and concentration of OA (p < 0.001). In July, 65% of the samples contained detectable levels of OA, compared with 38, 21 and 17%, in April, October and January respectively. Furthermore, 24% of the samples collected in July contained greater than 15 ng/ml of OA, while only 2, 9, and 1% of the samples collected in April, October and January respectively, contained greater than 15 ng/ml of OA. Based on the six samples collected from each herd, it appears that the presence and concentration of OA within a herd may be estimated from a limited number of animals per herd.