Peat and disinfectant powder used in swine husbandry systems - quantification of oral intake using toxic metals as potential markers.

The use of enrichment and bedding materials in pig husbandry intends to comply with the animals' behavioural needs to perform natural exploratory behaviour, which is strongly connected to foraging behaviour. It can thus be assumed that pigs will ingest a certain material quantity possibly posing a risk to animal health and food safety as previous studies identified contaminants in enrichment and bedding materials. However, risk assessment requires knowledge about the effective amount of ingested material. Voluntary material intake of pigs with free access to peat and disinfectant powder was estimated by measuring the tissue levels of toxic metals originating from the respective materials in 28 pigs (seven groups, n = 4) via inductively coupled plasma mass spectrometry and comparing the results to tissue levels of pigs fed with known amounts of metals. Additionally, as markers of consumption, n-alkanes and acid insoluble ash naturally occurring in the materials and titanium dioxide, added as an external marker to disinfectant powder, were analysed in pigs' faeces. Tissue levels of toxic metals as well as marker analyses in pigs' faeces could prove material consumption. Results revealed mean voluntary intake levels of peat and disinfectant powder by pigs up to 7% and 2% of the daily ration. Hence, a transfer of contained toxic metals into the food chain might occur. Although current maximum levels for toxic elements in animal tissues were not exceeded due to dietary inclusion of peat or disinfectant powder, dietary exposure through food of animal origin should be reduced to a possible minimum. This applies specifically for elements, where no health-based guidance values for humans could have been derived (e.g. arsenic). Thus, labelling guidelines for enrichment and bedding materials can be a perspective to limit the entry of toxic metals and trace elements into the environment.

Effects of 3-nitrooxypropanol and varying concentrate feed proportions in the ration on methane emission, rumen fermentation and performance of periparturient dairy cows.

The climate-relevant enteric methane (CH4) formation represents a loss of feed energy that is potentially meaningful for energetically undersupplied peripartal dairy cows. Higher concentrate feed proportions (CFP) are known to reduce CH4 emissions in cows. The same applies to the feed additive 3-nitrooxypropanol (3-NOP), albeit through different mechanisms. It was hypothesised that the hydrogen not utilised for CH4 formation through the inhibition by 3-NOP would be sequestered by propionate formation triggered by higher CFP so that it could thereby give rise to a synergistically reduced CH4 emission. In a 2 × 2-factorial design, low (LC) or high (HC) CFP were either tested without supplements (CONLC, CONHC) or combined with 3-NOP (NOPLC, 48.4 mg/kg dry matter (DM); NOPHC, 51.2 mg 3-NOP/kg DM). These four rations were fed to a total of 55 Holstein cows from d 28 ante partum until d 120 post partum. DM intake (DMI) was not affected by 3-NOP but increased with CFP (CFP; p < 0.001). CH4/DMI and CH4/energy-corrected milk (ECM) were mitigated by 3-NOP (23% NOPLC, 33% NOPHC) (p < 0.001) and high CFP (12% CON, 22% 3-NOP groups) (CFP × TIME p < 0.001). Under the conditions of the present experiment, the CH4 emissions of NOPLC increased to the level of the CON groups from week 8 until the end of trial (3-NOP × CFP × TIME; p < 0.01). CO2 yield decreased by 3-NOP and high CFP (3-NOP × CFP; p < 0.001). The reduced body weight loss and feed efficiency in HC groups paralleled a more positive energy balance being most obvious in NOPHC (3-NOP × CFP; p < 0.001). ECM was lower for NOPHC compared to CONHC (3-NOP × CFP; p < 0.05), whereas LC groups did not differ. A decreased fat to protein ratio was observed in HC groups and, until week 6 post partum, in NOPLC. Milk lactose and urea increased by 3-NOP (3-NOP; p < 0.05). 3-NOP and high CFP changed rumen fermentation to a more propionic-metabolic profile (3-NOP; CFP; p < 0.01) but did not affect rumen pH. In conclusion, CH4 emission was synergistically reduced when high CFP was combined with 3-NOP while the CH4 mitigating 3-NOP effect decreased with progressing time when the supplement was added to the high-forage ration. The nature of these interactions needs to be clarified.

The effect of crude protein content of the diet on renal energy losses in horses.

Renal energy losses of horses are high in comparison with other species. In the present study, more data were obtained on this parameter to improve predictive equations for renal energy losses. Four adult ponies (247-344 kg body weight [BW]) were fed with eight different diets based on first cut hay, second cut hay, early first cut fresh grass, late cut herbs-grass mix, early cut clover-grass mix, sugar beet pulp, rice bran and straw. Feed intake was measured, and urine and faeces were quantitatively collected for 3 × 12 hr at daytime and afterwards 3 × 12 hr at nighttime. Feed was analysed for crude nutrients, gross energy, amino acids and neutral-detergent-insoluble crude protein (NDICP); faeces were analysed for crude nutrients and gross energy; and urine was analysed for nitrogen (N) and gross energy. Renal energy losses per gram dry matter (DM) intake (y; kJ/g DM) were strictly correlated to protein content in DM (x; g/kg DM): y = 0.325 + 0.00431x; r2  = .81; n = 38; p < .001. The data suggest that the intercept represents energy losses by detoxification products such as hippuric acid and the regression coefficient by protein metabolites such as urea.

Effect of oral supplementation of probiotic strains of Lactobacillus rhamnosus and Enterococcus faecium on the composition of the faecal microbiota of foals.

Effects of probiotics on the intestinal microbiota of foals are yet insufficiently studied. The aim of this study was to investigate whether supplementation of Lactobacillus rhamnosus (DSM 7133) and Enterococcus faecium (DSM 7134) influences the bacterial composition of the faecal microbiota of foals. A total of 34 newborn foals were randomly assigned to the placebo group (PG, n = 16) and the treatment group (TG, n = 18). From day 1 to day 14 of life, foals orally received 3 ml of either a probiotic preparation (1.05 × 109 CFU E. faecium and 4.50 × 108 CFU L. rhamnosus) or placebo (carrier) once a day. Faeces were collected directly from the rectum immediately after birth (meconium) and at day 14 and day 56 of life. Samples of 12 foals per group were selected for microbiological analysis. DNA was extracted and used for polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and quantitative PCR. No DNA or amplicons were obtained from meconium. There were no differences in richness of bands and Shannon index of diversity regarding the Clostridium cluster XIVa between groups. Cluster analysis and principal coordinate analysis of DGGE data showed a clear effect of age. Band-based similarity of bacterial clusters (Dice coefficient) decreased from day 14 to day 56 of life (p < 0.001) in PG foals only resulting in lower similarity in PG versus TG foals when 2 month old (p < 0.01). Five of thirty re-amplified bands were identified on species level. Others were assigned either to family (mainly Lachnospiraceae) or genus level (Akkermansia). The bands related to Akkermansia muciniphila or Akkermansia spp. appeared almost in all DGGE profiles. Two-week supplementation of the probiotic preparation to foals had no significant impact on the composition of the faecal microbiota but it appears to have prevented the reduction of bacterial similarity between 2 and 8 weeks of age observed in not treated foals.

Effects of oral supplementation of probiotic strains of Lactobacillus rhamnosus and Enterococcus faecium on diarrhoea events of foals in their first weeks of life.

Foal first diarrhoea is one of the most prominent problems in the early life of horses. Probiotics might have the potency to prevent or at least diminish neonatal diarrhoea. We hypothesised that the treatment of foals with probiotic strains of Lactobacillus rhamnosus and Enterococcus faecium starting early after birth and then daily over 2 weeks would prevent or mitigate foal heat diarrhoea. The influence of this probiotic treatment on diarrhoea incidence and growth and health performance of young foals was investigated. Thirty-four foals were randomly allocated to two groups. From day 1 to 14 of life, the foals received either placebo (PG, n = 16) or the probiotic treatment (TG, n = 18). Clinical examination was performed, and the faeces consistency score (FCS, 1-5; with diarrhoea defined by ≤3) was recorded once per day in weeks 1 and 2 and once weekly in weeks 3-8 of life (WL). The body height was measured at birth and after two and eight WL. Diarrhoea occurred in the 1st WL in 19% and 61% of PG and TG foals respectively. In the 1st WL, diarrhoea lasted 0.3 ± 0.8 and 1.6 ± 1.4 days in PG and TG foals respectively. In the 2nd WL, diarrhoea occurred in 94% and 84% of PG and TG foals, respectively, and lasted for 3.0 ± 1.5 and 3.7 ± 1.6 days respectively. At least two periods of diarrhoea developed in 33% and 65% of PG and TG foals respectively. The TG foals grew slightly slower than the PG foals. The results indicated that the probiotic treatment of neonatal foals as performed in this study was not suitable to reduce diarrhoea within the first two WL, because contrary to the hypothesis, the TG foals suffered more frequently and for longer periods from diarrhoea than the PG foals.

Development and Evaluation of qPCR Assay for Quantitation of Kazachstania slooffiae and Total Yeasts Occurring in the Porcine Gut.

Kazachstania slooffiae is the dominating yeast in pig's gut. No methods others than cultivation were applied for enumeration of yeasts within this ecosystem. Therefore, the aim of this study was to develop a real-time quantitative polymerase chain reaction (qPCR) assay to quantitate total yeasts and K. slooffiae in the porcine gut. This work demonstrated that the copy numbers in gDNA can be determined by qPCR using PCR amplicons as a calibrator and one-point calibration method. The gDNA were then used as a calibrator for further analysis. The values of quantitation cycle and PCR amplification efficiency of gDNA calibrator were highly reproducible. DNA was extracted from feces and from 10 different cultured yeasts found in pigs' intestine. The qPCR results using primers NL1/LS2 encoding 26S rDNA correlated (r = 0.984, P < 0.0001) with cultivation results. From two primer sets developed, one set encoding act1 gene was suitable for quantitation of K. slooffiae. The copy numbers of K. slooffiae could be determined by 40% analyzed animals, amounting to about 70% of total yeasts. The application of this method in next studies will help to get more information about K. slooffiae and total yeasts in the gut of pigs.

Higher body fatness in intrauterine growth retarded juvenile pigs is associated with lower fat and higher carbohydrate oxidation during ad libitum and restricted feeding.

PURPOSE: A thrifty energy metabolism has been suggested in intrauterine growth restricted (IUGR) offspring. We characterized energy metabolism and substrate oxidation patterns in IUGR pigs in response to food restriction (FR) and refeeding (RFD). METHODS: Female pigs with low (L; 1.1 kg; n = 20) or normal birth weight (N; 1.5 kg; n = 24) were fed ad libitum after weaning. Half of L and N pigs were food restricted (R; LR, NR) from days 80 to 100 (57% of ad libitum) and refeed from days 101 to 131, while the remaining pigs were fed ad libitum (control, C). Using indirect calorimetry, carbohydrate and fat oxidation (COX, FOX), energy expenditure (EE) and balance (EB), resting metabolic rate (RMR) [all related to kg body weight(0.62) (BW)] and RQ were determined at 4 days before (day 76) and after (day 83) beginning of FR, 4 days before (day 97) and after (day 104) end of FR and 25 days after beginning of RFD (day 125). Body fat and muscle weights were determined at day 131. RESULTS: In spite of higher relative food intake (FI), BW was lower in L pigs. In L pigs, physical activity was lower at age 76 and 83 days compared to N pigs. IUGR did not affect EE or RMR, but resulted in higher COX and lower FOX, causing greater and earlier onset of fat deposition. During FR, EE and RMR of R pigs dropped below that of C pigs, and BW gain was delayed by 30% irrespective of birth weight. In response to FR, COX decreased and FOX increased. During FR, in LR pigs FOX was ~50% of that in NR pigs. After 4 days, but not 25 days of RFD, EB and fat synthesis were higher in pigs previously subjected to FR, indicating early catch-up fat. In R pigs, BW and the abdominal fat proportion were lower at 131 days. CONCLUSIONS: Differences in food intake and substrate oxidation pattern, but not in EE and RMR, between L and N pigs were reflected in higher body fat proportions but lower body and muscle weights in L pigs. Refeeding following FR was initially associated with increased FI, a more positive EB and a more intense stimulation of fat synthesis which did not persist after 25 days of refeeding.

Effect of ensiling treatment on secondary compounds and amino acid profile of tropical forage legumes, and implications for their pig feeding potential.

BACKGROUND: Smallholders in the tropics depend on local protein supplements to balance pig diets. Thus, various tropical forage legumes are a potential feeding option. Ensiling allows converting forages into a ready-to-feed-out choice, but the lactic acid fermentation may influence various (anti)nutritional components. The purpose of this study was to evaluate the effects of sucrose (SU) and a tropical Lactobacillus plantarum strain (LAB) as ensiling treatments (control, SU, LAB, LAB + SU) on the potential nutrient availability of 10 forage legume species. RESULTS: Ensiling commonly reduced antinutritional compounds such as tannins (by 49-84%) and trypsin inhibitory activity (by 74-78%), as well as oxalic acid (by 51-100%). An improved potential absorbability of protein and minerals for pigs is thus inferred. There was no major loss in total amino acids. In general, the species effect was stronger than the treatment effect. A clear effect of the treatments SU, LAB and LAB + SU over all 10 forage species was only observed for oxalic acid, single amino acids and trypsin inhibitory activity. CONCLUSION: Ensiling is a viable option to enhance nutrient utilization of tropical forages for pigs. Species-specific treatment of forage legumes is recommended.

In vitro ruminal degradability of wheat straw cultivated with white-rot fungi adapted to mushroom farming conditions.

Biological treatment of cereal straw for ruminant nutrition purposes might present an environmentally friendly option of valorizing a widely available by-product of grain production for farming systems with low external input. Several strains of white-rot fungi have been selected in the past under mostly controlled laboratory conditions for their capacity of lignin degradation. The study adapted to conditions on farm for upscaling purposes. The development of the in vitro straw digestibility with two different moistening pre-treatments and inoculated with three different fungi species, namely Pleurotus ostreatus, Ceriporiopsis subvermispora and Volvariella volvacea, was determined up to 42 days of fermentation with five sampling times. The effect of physical straw pre-treatments on nutritional parameters was evaluated. The neutral detergent fiber digestibility (NDFD30h), enzymatically soluble organic substance (ELOS) and the gas production (Hohenheim Feed value Test, HFT) as indicators for in vitro ruminal degradability decreased over time independent of the fungus: HFT, ELOS and NDFD30h by up to 50, 35 and 30% of the original straw. Remoistening and autoclaving the straw increased the gas production significantly by 2.6 mL/200 g dry matter (DM), and ELOS and NDFD30h by 45 and 51 g/kg DM compared to the original straw (34.9 mL/200 mg DM, 342 g/kg DM, 313 g/kg NDF).

Feed Clusters According to In Situ and In Vitro Ruminal Crude Protein Degradation.

Effective degradation (ED) of crude protein (CP) was estimated in vitro at 0.02, 0.05 and 0.08 h−1 assumed ruminal passage rates for a total of 40 feedstuffs, for which in situ ED was available and used as reference degradation values. For this, the Streptomyces griseus protease test was used. The differences between in vitro CP degradation and the in situ CP degradation values were lowest in legume grains and highest in cereal by-products and barley. The differences between in situ and in vitro ED were expressed using a degradation quotient (degQ), where degQ = (EDin vitro − EDin situ)/EDin situ. Among the tested feedstuffs, eight specific clusters were identified according to degQ for the assumed passage rates. The feedstuffs clustered in an unspecific way, i.e., feedstuffs of different nutrient composition, origin or treatment did not necessarily group together. Formaldehyde−treated rapeseed meal, soybean meal, wheat, a treated lupin, sunflower meal and barley could not be assigned to any of the clusters. Groupwise degradation (range of degQ for assumed passage rates are given in brackets) was detected in grass silages (−0.17, −0.11), cereal by-products together with sugar beet pulp (−0.47, −0.35) and partly in legume grains (−0.14, 0.14). The clustering probably based on different specific nutrient composition and matrix effects that influence the solubility of feed protein and limit the performance of the protease. The matrix can be affected by treatment (chemically, thermally or mechanically), changing the chemical and physical structure of the protein within the plant. The S. griseus protease test had reliable sensitivity to reflect differences between native feedstuffs and treatments (thermally or chemically) that were found in situ. The in situ results, however, are mostly underestimated. The clustering results do not allow a clear conclusion on the groupwise or feed-specific use of carbohydrate-degrading enzymes as pre- or co-inoculants as part of the S. griseus protease test and need to be tested for its potential to make this test more conform with in situ data.

Preference and possible consumption of provided enrichment and bedding materials and disinfectant powder by growing pigs.

BACKGROUND: Domestic pigs have an evolutionary conserved exploratory behaviour. To comply with this requirement, the European Union aims at setting standards for appropriate enrichment materials for pigs (Council Directive 2008/120/EC). As recommended characteristics include 'chewable' and 'edible', pigs might also consume these materials (Commission Recommendation (EU) 2016/336), which are often additionally advertised to enhance lying comfort and hygienic conditions in stables. To date, a wide range of bedding, enrichment and disinfectant materials is available on the market to ensure environmental enrichment, a dry, hygienic environment or lying comfort. Previous studies revealed considerable amounts of undesirable substances in some of these materials possibly being a risk for food safety considering oral uptake by the animal. To determine interest and indicators for consumption of different types of materials by pigs during exploratory behaviour, a camera-assisted observational study with 12 female pigs (German Landrace) was conducted. We tested their preference for a disinfectant powder, peat, biochar and straw as reference material in a 4 × 6 factorial arrangement. RESULTS: Pigs manipulated and consumed all offered materials. However, longest manipulation time per pig was observed for biochar (63 min/day) and peat (50 min/day) (p < 0.05). Analyses of the bulk molecular-chemical composition and n-alkanes and acid insoluble ash as markers in the materials and in faeces clearly revealed the consumption of these materials by pigs. CONCLUSIONS: Whether the consumption of considerable amounts together with certain levels of undesirable substances represents a risk for pig and consumer health could yet not be established. Future studies will address the quantitative contribution of undesirable substances by oral ingestion of bedding and enrichment materials and disinfectant powders to the daily feed ration.

Effect of Hay Steaming on the Estimated Precaecal Digestibility of Crude Protein and Selected Amino Acids in Horses.

Steaming hay is increasingly used to treat low-quality forage because it was proven to reduce inhalable allergens such as mould spores, bacteria, and airborne dust particles. Preliminary results have shown a substantial loss of precaecal (pc) digestibility (D) of crude protein (CP) and amino acids (AA). For this purpose, six different batches of hay from central Germany were divided into four subsamples, and each one was individually steamed. Native hay and four replicates of each steamed subsample were analysed for CP, AA, neutral detergent insoluble crude protein (NDICP), neutral detergent soluble crude protein (NDSCP) as well as pepsin insoluble CP (piCP). Based on the analytical parameters, pcD of CP, protein solubility (PS), piCP (% CP) and precaecal digestible (pcd) CP and pcdAA contents were calculated. Selected Maillard reaction products (MRP), namely furosine and carboxymethyllysine (CML), were also analysed. Steaming did not affect CP content (native = 69, steamed = 67 g/kg dry matter, DM; p > 0.05), but it had an impact on the insoluble part of CP. Thus, NDICP increased by 57% (native = 27, steamed = 42 g/kg DM; p < 0.05) and piCP by 15% overall (native = 40, steamed = 46% of CP; p < 0.05). This could be a consequence of the heat damage and the associated increase in MRP. The content of furosine rose by 67% (native = 17.6, steamed = 29.4 mg/100 g DM; p < 0.05). The content of CML increased by 120% (native = 5.1, steamed = 11.3 mg/100 g DM; p < 0.05). We chose to analyse these two MRPs because they represent the reaction products with the limiting AA lysine. In contrast, the soluble fractions of CP declined, while PS as a percentage of CP decreased by 38% as a result of the treatment, and NDSCP was reduced by as much as 41% (p < 0.05). In line with this, the steaming process decreased the pcD of CP (native = 56%, steamed = 35%; p < 0.05) and pcdCP (native = 37.9, steamed = 22.5 g/kg DM; p < 0.05), respectively. The same effects were shown for selected AA; e.g., sulphuric AA pcd methionine plus pcd cysteine decreased by 45%, pcd threonine decreased by 41%, and the limited AA pcd lysine decreased by more than 50% (p < 0.05). In conclusion, the high temperatures generated during steaming lead to protein damage and consequently to a reduction in the pcD of CP and essential AA. Nevertheless, steaming successfully reduces viable microorganisms and binds dust particles. Therefore, steamed hay is still a proper and sometimes the only possible roughage for horses suffering from respiratory diseases such as equine asthma. Essentially, horse diets based on steamed hay should be balanced accordingly.

Physicochemical characterisation of barley straw treated with sodium hydroxide or urea and its digestibility and in vitro fermentability in ruminants.

The development of strategies to overcome the shortage of forage due to persistently low rainfall is becoming a central task for animal nutrition in research and practice. In this study, it was investigated how the treatment of straw with NaOH or feed urea in a practicable procedure for modern farms affects rumen fermentation (gas production and greenhouse gas concentration) as well as the digestibility of feed energy and nutrients. For this purpose, the treatments were tested individually and in different proportions in a total mixed ration (TMR) in ruminal batch cultures in vitro and in a digestibility trial with sheep. In order to explain the observed effects at the molecular level, descriptive data from 13C solid state nuclear magnetic resonance (NMR) and Fourier-transform infrared (FTIR) spectroscopy were obtained. NaOH treatment of straw increased crude ash (CA), non-fibrous carbohydrates, digestible energy (DE), and metabolizable energy (ME) concentration, whereas the proportion of neutral detergent fibre (aNDFom) and hemicellulose decreased. In urea treated straw, NH3-N and crude protein increased, whereas acid detergent lignin (ADL), DE, and ME decreased. The physically effective fibre (peNDF8) concentration increased in TMR containing 18% of NaOH or urea treated straw (p < 0.01). The application of straw treatments as pure substrates (not as part of a TMR) increased gas production and decelerated ruminal fermentation (p < 0.05). In vitro organic matter digestibility (IVOMD) of the straw (0.31) increased after NaOH (0.51; p < 0.05) and urea treatment (0.41; p > 0.05). As part of a TMR, straw treatments had no distinct effect on gas production or IVOMD. Concentrations of CH4 and CO2 were likewise not affected. Apparent total tract digestibility of aNDFom, acid detergent fibre (ADFom), hemicellulose, and cellulose increased in the TMR by approximately 10% points following NaOH treatment (p < 0.05). The inclusion of urea treated straw did not affect apparent digestibility. Calculated true digestibility of aNDFom was 0.68, 0.74, and 0.79, of ADFom 0.58, 0.57, and 0.65, and of ADL 0.02, 0.13, and 0.08 in TMR including untreated, NaOH treated, and urea treated straw, respectively. 13C NMR and FTIR analyses consistently revealed that the global structure and crystallinity of the carbohydrates (cellulose and hemicellulose) was not altered by treatment and the concentration of lignin was likewise not affected. Depolymerisation of lignin did not occur. However, NMR signals assigned to acetyl groups were significantly altered indicating that straw treatments disrupted linkages between hemicelluloses and lignin. Moreover, the acetates signal was affected. This signal can be assigned to linkages between ferulic acids and hemicelluloses (arabinoxylans). FTIR spectra of straw treatments mainly differed at a wavelength of 1730 cm-1 and 1240 cm-1. Disappearance of the 1730 cm-1 peak suggests removal of hemicelluloses or lignin related compounds by treatment. The disappearance of the lignin peak at 1240 cm-1 could be due to conjugated ketone (phenyl-carbonyl) removal or the removal of ferulic and p-coumaric acid acetyl groups. Both treatments are supposed to release fermentable cell wall components (hemicelluloses) from lignin-associated bonds and as a result, straw fibre can be better fermented in the rumen. This contributes to energy supply and increased fibre digestibility at least in the TMR that contained NaOH treated straw. The alkaline straw treatments probably induced a release of phenolics such as ferulic acid and p-coumaric acid, which can be metabolised in the gut and the liver and metabolites might be excreted with the urine. This could notably contribute to metabolic energy losses.

Changes in Composition and Diversity of Epiphytic Microorganisms on Field Pea Seeds, Partial Crop Peas, and Whole Crop Peas during Maturation and Ensiling with or without Lactic Acid Bacteria Inoculant.

The present study was conducted under the hypothesis that, in field peas, type of plant material, stage of maturity, ensiling, silage additive, and aerobic stress affect the composition and diversity of epiphytic microbial communities. Epiphytic microbial composition and diversity of pea seeds, partial crop peas, and whole crop peas was analyzed at different stages of late maturity, before and after ensiling, and with or without the use of lactic acid bacteria (LAB) as inoculant. Suitable combinations among pea crop variants, maturity stages, and inoculant use for the production of stable silages with sufficient aerobic stability after opening and during feed-out were identified. Genomic DNA was extracted, and 16S and 18S rRNA gene amplicons were sequenced. To assess the quality of the various silages, nutrient concentration, pH value, concentration of lactic acid, short chain fatty acids, and alcohols, and aerobic stability were determined. Pea seeds were barely colonized by epiphytic microorganisms. In partial and whole crop peas, composition and α-diversity (Shannon index) of bacterial communities did not differ between crop variants but differed among maturity stages. Epiphytic eukaryotes were rarely found on partial and whole crop peas. Bacterial composition and α-diversity were affected by ensiling and subsequent aerobic storage. In partial and whole crop peas, plant maturation caused an increase of the relative abundance of naturally occurring LAB (Weissella, Pediococcus, and Lactobacillus spp.). As a possible result, natural LAB support stable ensiling conditions even without the use of inoculants beginning with a maturity of 78 on the BBCH scale. This corresponded with a dry matter (DM) concentration of 341 and 363 g/kg in partial and whole crop peas, respectively. Addition of LAB inoculants, however, reduced ammonia, acetic acid, and butanol concentrations, and supported aerobic stability. Earlier stages of plant maturity (BBCH 76 and 77, 300 g DM/kg or less) were more prone to microbial spoilage. Stable pea seed silages can be produced at a maturity between BBCH 78 (427 g DM/kg) and 79 (549 g DM/kg), but they undoubtedly require LAB inoculation or application of other ensiling agents. IMPORTANCE Field peas are important protein suppliers for human and animal nutrition. They can be grown in many areas of the world, which may reduce imports of protein plants and has beneficial economic and ecological effects. Ensiling is a method of preserving feed that can be implemented easily and cost-effectively at the farm. Peas harvested as seeds, partial crop, or whole crop at different maturities enable a wide range of applications. The study characterized epiphytic microbial communities on peas in terms of composition and diversity depending on the maturity of the plants and feed conservation by ensiling as they play an essential role for the production of silages. Even if this study did not consider year, site, or cultivar effects, the results would show which part of the plant is probably well suited for the production of stable and high-quality silages and at which stage of maturity.

Comparative evaluation of laboratory-scale silages using standard glass jar silages or vacuum-packed model silages.

BACKGROUND: The objective of this study was to compare the fermentation variables of laboratory-scale silages made in glass preserving jars (GLASS) and vacuum-packed plastic bags (Rostock model silages, ROMOS). Silages were prepared from perennial ryegrass (fresh and wilted, 151 g kg(-1) and 286 g kg(-1) dry matter (DM), respectively) and remoistened coarsely ground rye grain (650 g kg(-1) DM) either with or without the addition of a lactic acid bacteria inoculant (3×10(5) colony forming units (cfu) g(-1) , LAB). Quintuplicate silos were opened on days 2, 4, 8, 49 and 90. RESULTS: Silage pH (P=0.073), acetic acid content (P=0.608) and ethanol content (P=0.223) were not influenced by the ensiling method. The contents of DM (P<0.001) and propionic acid (P=0.008) were affected by the ensiling method, but mean differences were only marginal. In ROMOS the concentration of lactic acid was increased (P=0.007) whereas butyric acid was produced less (P=0.001) when compared to GLASS. This suggested slightly better ensiling conditions for ROMOS. CONCLUSIONS: ROMOS represents a reasonable alternative to glass jar silages and opens the possibility for further investigations, e.g. studying the impact of packing density as well as the quantitative and qualitative analysis of fermentation gases.

Hypoglycin A in Cow's Milk-A Pilot Study.

Hypoglycin A (HGA) originating from soapberry fruits (litchi, and ackee) seeds or seedlings from the sycamore maple (SM) tree (related to Sapindaceae) may cause Jamaican vomiting sickness in humans and atypical myopathy in horses and ruminants. A possible transfer into dairy cow's milk cannot be ruled out since the literature has revealed HGA in the milk of mares and in the offal of captured deer following HGA intoxication. From a study, carried out for another purpose, bulk raw milk samples from four randomly selected dairy farms were available. The cows were pastured in the daytime. A sycamore maple tree was found on the pasture of farm No. 1 only. Bulk milk from the individual tank or milk filling station was sampled in parallels and analyzed for HGA by LC-ESI-MS/MS. Measurable concentrations of HGA occurred only in milk from farm No. 1 and amounted to 120 and 489 nmol/L. Despite low and very variable HGA concentrations, the results indicate that the ingested toxin, once eaten, is transferred into the milk. However, it is unknown how much HGA the individual cow ingested during grazing and what amount was transferred into the bulk milk samples. As a prerequisite for a possible future safety assessment, carry-over studies are needed. Furthermore, the toxins' stability during milk processing should also be investigated as well.

Methylenecyclopropylglycine and hypoglycin A intoxication in three Pére David's Deers (Elaphurus davidianus) with atypical myopathy.

BACKGROUND: Hypoglycin A (HGA) and methylenecyclopropylglycine (MCPrG) from seeds/seedlings of Sycamore maple (SM, Acer pseudoplatanus) causes atypical myopathy (AM) in horses. AM was not known to occur in wild ruminants until several fatalities in milus (Elaphurus davidianus) following the ingestion of HGA in SM seeds. However, a role for MCPrG has not previously been evaluated. OBJECTIVES: To test the hypothesis that MCPrG is also a major factor in AM in milus, three milus (M1, M2, M3) from the Zoo Dresden (aged 7-11 years, 2 females and 1 male, in good nutritional condition) that developed AM were studied. METHODS: Serum, urine and methanol extracts from the liver, kidney, rumen digesta and faeces were analysed by ultrahigh-performance liquid chromatography-tandem mass spectrometry for HGA, MCPrG and for conjugates of carnitine (C) and glycine (G): Methylenecyclopropylacetyl (MCPA)-G, MCPA-C, Methylenecyclopropylformyl (MCPF)-G, MCPF-C, butyryl-C and isobutyryl-C. RESULTS: HGA in serum was high (M2 480 nmol/L; M3 460 nmol/L), but MCPrG was not. HGA and MCPrG were found in rumen and faeces extracts, and MCPrG was also identified in the liver. Metabolites of HGA and MCPrG were high in serum, urine and liver, but not in the rumen or faeces. CONCLUSIONS: This study shows that MCPrG is involved in the pathophysiology of AM in milus. The metabolism of MCPrG is considered to be faster because, after ingestion, the specific metabolites appear highly concentrated in the serum. The high toxin concentration in the liver suggests that a possible transfer into products for human consumption may pose a risk.

Fermentation Characteristics and In Vitro Digestibility of Fibers and Fiber-Rich Byproducts Used for the Feeding of Pigs.

Dietary fibers may have positive impact on health and wellbeing of pigs. The study examined physicochemical properties of two lignocelluloses (including and excluding bark), powdered cellulose, Aspergillus niger mycelium, lucerne chaff, soybean shells, wheat bran, and sugar beet pulp in relation to fermentability and digestibility using in vitro batch-culture incubation. Maize starch and a purified cellulose were used as standardized substrates for classification of the test substrates. The substrates covered a wide range regarding their physicochemical properties. Swelling capacity (SC) was 9-411%, water binding capacity (WBC) was 4.4-14.3 g/g dry matter (DM), and water holding capacity (WHC) was 4.1-10.6 g/g DM. Gas production and other fermentation parameters-namely post-incubation pH, CH4, NH3, and short chain fatty acids (SCFA) concentrations-revealed a significant fermentation of sugar beet pulp, soybean shells, lucerne chaff, wheat bran, A. niger mycelium, and powdered cellulose, whereas the lignocelluloses were not fermented. Significant correlations were found between the physicochemical properties and the fermentation parameters (p < 0.05). Enzymatic pre-digestion mostly reduced gas, NH3, and SCFA production. In vitro digestibility of DM (IVDMD) and organic matter (IVOMD) was mostly negligible after enzymatic pre-digestion. Fermentation alone led to only 0.10-0.15 IVDMD and 0.14-0.15 IVOMD in lignocelluloses and powdered cellulose, respectively, but 0.44-0.37 IVDMD and 0.46-0.38 IVOMD in the remainder of substrates (p < 0.05). In vitro digestibility was again correlated with the physicochemical properties of the substrates and the fermentation parameters (p < 0.05). The fiber preparations and fiber-rich byproducts were fermented to a relevant extent. In contrast, lignocelluloses were not fermented and can be used rather as bulk material.

Feed Intake Parameters of Horses Fed Soaked or Steamed Hay and Hygienic Quality of Hay Stored following Treatment.

Horses suffering from equine asthma must consume low-dust forage, with soaking and steaming being suitable methods of hay treatment. The impacts of this treated hay's subsequent storage and effects on the horses' chewing activity are largely unknown. Meadow hay was soaked (10-15 °C, 15 min) or steamed (100 °C, 60 min). Microbial counts (colony forming units (CFU)) were determined by culture before and after soaking or steaming, and subsequent storage at 10 and 25 °C for 6, 12 and 24 h (three replicates each). Six horses were fed native, soaked and steamed hay, according to a cross-over design, and chewing parameters were measured. Steaming reduced (p < 0.05) typical mold vs. soaking (0 vs. 50 CFU/g) and yeasts vs. native and steamed hay (0 vs. 102 and 90 CFU/g). Storing soaked hay elevated bacteria, mold, and yeasts (p < 0.05). Within the first 60 min of hay intake, the steamed hay and soaked hay were eaten slower (19.5 and 21.5 g dry matter/min, respectively; p < 0.05) and the steamed hay was chewed more intensely (steamed hay: 3537; native: 2622; and soaked: 2521 chewing cycles/kg dry matter, p < 0.05). Steaming particularly improves the hygienic quality of hay. Soaked hay is not stable when stored and is less accepted by horses.

Degradation of Monosaccharides, Disaccharides, and Fructans in the Stomach of Horses Adapted to a Prebiotic Dose of Fructooligosaccharides and Inulin.

For a period of 20 days, 12 horses either received a prebiotic supplementation with fructooligosaccharides and inulin via Jerusalem artichoke meal (JAM) or corncob meal without grains (CMG) as placebo. The horses were euthanized 1 hour postprandial, gastric digesta was sampled from pars nonglandularis (PNG) and pars glandularis (PG), and concentrations of starch, mono- and disaccharides, fructans, d- and l-lactic acid, and short chain fatty acids were analyzed. Concentrations of starch and simple sugars were widely the same in JAM supplemented and not supplemented meals. However, fructans were less than half as much without supplementation as with supplementation of JAM. Glucose, fructose, sucrose, and fructans disappeared to a larger extent with prebiotic supplementation than without (106.6% vs. 86.7% glucose, 73.1% vs. 66.8% fructose, 91.5% vs. 14.7% sucrose, and 68.3% vs. 35.4% fructans remained in PNG; 81.9% vs. 38.3% glucose, 52.2% vs. 53.4% fructose, 47.1% vs. 0% sucrose, and 48.5% vs. 31.7% fructans remained in PG with CMG vs. JAM feeding). Disappearance of simple sugars and fructans was primarily associated with appearance of n-butyric acid (r = -0.21 - r = -0.33).