The Ruminant sorting mechanism protects teeth from abrasives.

Dental wear due to ingestion of dust and grit has deleterious consequences. Herbivores that could not wash their food hence had to evolve particularly durable teeth, in parallel to the evolution of dental chewing surface complexity to increase chewing efficacy. The rumen sorting mechanism increases chewing efficacy beyond that reached by any other mammal and has been hypothesized to also offer an internal washing mechanism, which would be an outstanding example of an additional advantage by a physiological adaptation, but in vivo evidence is lacking so far. Here, we investigated four cannulated, live cows that received a diet to which sand was added. Silica in swallowed food and feces reflected experimental dietary sand contamination, whereas the regurgitate submitted to rumination remained close to the silica levels of the basal food. This helps explain how ruminants are able to tolerate high levels of dust or grit in their diet, with less high-crowned teeth than nonruminants in the same habitat. Palaeo-reconstructions based on dental morphology and dental wear traces need to take the ruminants' wear-protection mechanism into account. The inadvertent advantage likely contributed to the ruminants' current success in terms of species diversity.

The effect of size and density on the mean retention time of particles in reindeer (Rangifer tarandus).

Particle passage from the reticulorumen (RR) depends on particle density and size. A classic way of assessing these effects is the use of plastic markers of varying density and size that are recovered in the faeces. Here, we report results of an experiment where four fistulated reindeer (Rangifer tarandus, 96 ± 12 kg) were fed two different diets (browse, voluntary dry matter intake [DMI] 70 ± 10 g/kg0.75/d; or a pelleted diet, DMI 124 ± 52 g/kg0.75/d) and dosed via fistula with 8 different particle types combining densities of 1.03, 1.22 and 1.44 g/ml and sizes of 1, 10 and 20 mm. Generally, particles that passed the digestive tract intact (not ruminated) did so relatively early after marker dosing, and therefore had shorter mean retention times (MRT) than ruminated particles. On the higher intake, the overall mean retention time (MRT) of particles was shorter, but this was not an effect of shorter MRT for either intact or ruminated particles, but due to a higher proportion of intact particles at the higher intake. This supports the concept that ruminants do not adjust chewing behaviour depending on intake, but that a lower proportion of digesta is submitted to rumination due to pressure-driven escape from the forestomach at higher gut fills. Compared to cattle (Bos primigenius taurus), muskoxen (Ovibos moschatus) and moose (Alces alces) that had received the same markers, reindeer had a lower proportion of 1 mm particles that passed intact. Our results support the concept that the critical size threshold for particles leaving the ruminant forestomach is dependent on body size. While the results likely do not represent findings peculiar for reindeer, they indicate fundamental mechanisms operating in the forestomach of ruminants.

Comparative study of feeding and rumination behaviour of goats and sheep fed mixed grass hay of different chop length.

Rumination is reported to be more pronounced in sheep compared to goats. This study compared the feeding and rumination behaviour of small ruminants and consisted of two experiments (E1 and E2). In E1, four sheep and four goats were offered low-quality hay (NDFom: 692 g/kg dry matter [DM]), processed to two chop lengths (long hay [LH]: 35 mm; short hay [SH]: 7 mm) in a 2 × 2 factorial (2 species × 2 chop lengths), cross-over design. In E2, the same animals were offered moderate-quality hay (NDFom: 636 g/kg DM) processed as LH and SH. Hay was offered for ad libitum consumption. Feeding and rumination behaviour was evaluated using video recordings. Aspects of rumination like chewing frequency were evaluated for 30 min per day. Faecal samples were analysed for faecal-N and particle size. There was no species effect on feed intake and organic matter digestibility (faecal N as proxy); however, goats consumed more LH than SH in E1 and E2. There was an effect of species on rumination:eating duration (R:E) ratio (higher in sheep) in E1 but not in E2, where there was a tendency for a species effect on rumination duration. In E1 and E2, sheep had a higher R:E ratio for SH than for LH. For rumination behaviour, there was a species effect for number of daily boli, chewing frequency and chews per day (more in sheep) in E1 and E2. No effect of species was found for faecal particle size. Despite much concordance, feed comminution behaviour differed in some aspects between sheep and goats. In an evolutionary context, a shift of significance of rumination could be triggered by a higher amount of abrasives in natural diets of sheep, rendering a shift of chewing towards ruminally prewashed material a rewarding strategy.

Is there a difference in ruminal fermentation control between cattle and sheep? A meta-analytical test of a hypothesis on differential particle and fluid retention.

Ruminant species differ in digestive physiology. The species-specific ratio of mean retention time of particles and fluid (MRTparticle/MRTfluid) in the reticulorumen has been interpreted as controlling ruminal fermentation: a higher ratio indicates of a more distinct 'washing' of particulate digesta by liquid. This should increase the harvest of microbes from the reticulorumen, and keep the microbiome in a state of more intense growth; at the same time, this should increase the metabolic losses of faecal nitrogen of microbial origin, leading to lower values for the apparent digestibility of crude protein (aD CP). A systematic difference has been hypothesized between cattle (higher ratio) and sheep (lower ratio), with a lower MRTfluid in cattle due to a higher saliva production. Here, we test these hypotheses in a meta-analysis, using only studies that investigated cattle and sheep simultaneously. The datasets included 12 studies on MRT (of which 11 contained information on feed intake), yielding 102 (or 89) individual data; and 26 studies on protein digestibility (of which 18 contained information on intake), yielding 349 individual data. Cattle had a higher MRTparticle/MRTfluid (2.1) than sheep (1.7), mainly due to longer MRTparticle; only if body mass was included in the model, MRTfluid was significantly shorter in cattle in the larger MRT dataset (and tended to be shorter in the slightly smaller dataset). Cattle had a significantly lower aD CP than sheep, while there was no such difference in overall (dry or organic matter) digestibility. The dataset confirms a shift in fermentation strategy towards microbial production in cattle. While this has been suggested for ruminants in general, cattle appear particularly far on an evolutionary trajectory of maximizing microbial yield from the forestomach. The application of more specific digestive physiology data (like endogenous losses) gained from sheep to cattle should be done bearing these differences in mind.

Relationship between reticular pH parameters and potential on-farm indicators in the early lactation of dairy cows.

Subacute ruminal acidosis (SARA) is an important nutritional disorder affecting animal welfare and economy of milk production. Definitions rely on ruminal pH but due to limitations of its measurement, indicators reflecting low pH are highly desirable. The aim of this study was to investigate the relationship between reticular pH and 18 on-farm indicators in milk, blood, faeces, urine and chewing behaviour in early lactating dairy cows. Ten farms were visited for 3 weeks and in total samples of 100 cows (10 per farm) were taken. The statistics and graphical visualization were performed using Pearson correlation and linear regression models on an animal individual level as well as with linear mixed models. Eight indicators (milk fat, fat-to-protein ratio, rumination time, feed intake time, rumination frequency, rumination boluses, lying time and faecal pH) were statistically significant associated with the daily animal individual reticular pH average. However, none of the models including the potential explanatory variables explained more than 5% of the pH variations. The study confirms the necessity of pH measurement to detect SARA risk animals in early lactation dairy cows.

Individual differences in digesta retention and their relation to chewing in cattle-A pilot investigation.

While information on individual differences in digesta mean retention time (MRT) might be interesting when selecting phenotypes for digestive efficiency, MRT measurements are prohibitively labour-intensive for large-scale application. Therefore, more easily measured proxies of MRT might be helpful. We used the opportunity of an experiment applying saliva stimulant in cattle to investigate the effect of different individual chewing behaviour on fluid and particle MRT with a consistent diet. Four non-lactating cattle (670-850 kg body mass [BM]) were used in a 4 × 4 Latin square design, treated with the saliva stimulant pilocarpine in dosages of 0, 1, 2.5 and 5 mg/kg BM per day. The cattle were fed hay with dry matter intake (DMI) assigned according to their metabolic body weight. MRT in the whole gastrointestinal tract (GIT), the reticulorumen (RR) and the distal tract were measured using Co-EDTA, Cr-mordanted fibre and La-mordanted fibre as markers representing fluid, small particles (2 mm) and large particles (1 cm), respectively. The chewing behaviour was measured via noseband pressure sensor and expressed as chewing frequency (chews per time) and chewing intensity (chews per DMI), both for total chewing (ingestion plus rumination) and rumination chewing alone. The animals differed considerably in chewing behaviour and MRT measures. BM did not show a significant effect on chewing behaviour and MRT measures, though it tended to negatively correlated to total chewing intensity. Chewing intensity exerted a significant negative influence on MRT of fluid and particles in the RR, which was not the case for chewing frequency. Chewing frequency showed a significant relationship with MRT of large particles in the GIT. We suggest that chewing behaviour could influence MRT in two ways: (i) by affecting saliva production via the masticatory-salivary reflex and subsequently, the fluid inflow to the RR; (ii) by contributing to particle size reduction. Should the link between chewing behaviour and MRT be corroborated in larger studies, chewing measures, with their large interindividual variation, could emerge as an easy-to-measure proxy for MRT characteristics.

Effect of induced saliva flow on fluid retention time, ruminal microbial yield and methane emission in cattle.

Both in vitro and animal studies indicated that a higher dilution rate is related to a more efficient microbial synthesis and a lower methane (CH4 ) yield. The latter could be a consequence of the former, as an increase in microbial cell synthesis offers an alternative hydrogen sink competing with methanogenesis. To test this assumption in live animals, we applied a saliva stimulant, pilocarpine, to modify liquid flow rate in cattle. Four non-lactating cows (750 ± 71 kg) were fed forage only (restricted to constant intake) in a 4 × 4 Latin square design with oral doses of 0, 1, 2.5 and 5mg pilocarpine/kg body weight and day. We quantified feed and water intake, ruminal and total tract mean retention time (MRT) of solute and particle markers, ruminal microbial yield (via urinary purine bases or metabolic faecal nitrogen), CH4 emission, digestibility, chewing behaviour, reticular motility and rumen fluid parameters. The effect of induced saliva flow was evident by visibly increased salivation and water intake. Increasing the pilocarpine dosages resulted in a linearly decreased MRT of fluid and small particles (p < 0.001 and p< 0.05, respectively) and methane yield as related to digested DM (p < 0.05), the latter at a magnitude of 5%. No effect of treatment was found on ruminal microbial yield estimated via purine derivates. Metabolic faecal N as an indicator of microbial growth linearly correlated with pilocarpine dosages (p < 0.05). No significant relationship was found between pilocarpine dosages and large particle MRT, nutrient digestibility, ruminal pH and short-chain fatty acids. In conclusion, different from some in vitro studies, there was little indication of a reciprocal effect of CH4 and microbial biomass production in cows fed a forage-only diet.

A pilot investigation on the effect of induced saliva flow on digestive parameters in sheep, and a comparison with cattle.

Sheep with a relatively low methane yield were observed to have shorter fluid and particle mean retention times (MRT). Because the application of pilocarpine, a saliva stimulant, was successful in reducing retention times in ruminants in previous studies, we applied this substance to sheep, expecting a reduction in MRT and methane yield. Three non-pregnant sheep (74 ± 10 kg) were fed a hay-only diet in a 3 × 3 Latin square design with oral doses of 0, 2.5 and 5 mg pilocarpine/kg body weight and day. Measurements included feed and water intake, MRT of liquid and particulate phases in the reticulorumen (RR) and total gastrointestinal tract (GIT), ruminal microbial yield (via urinary purine bases and metabolic faecal nitrogen), total tract methane emission, apparent nutrient digestibility and rumen fluid parameters. Data were investigated for linear and quadratic effects using orthogonal polynomial contrasts. The MRT of liquid and small particles in the RR and total GIT, and the short-chain fatty acid concentration in rumen fluid, linearly declined with increasing pilocarpine dosage, while no quadratic relationship was detected. Intake of feed DM and water, apparent nutrient digestibility, methane yield and microbial yield were not affected by pilocarpine. When combining the sheep data with that of a similar experiment in cattle, we found that the MRT of the liquid phase was positively associated with estimated NDF digestibility and with methane production per digested NDF, but was not associated with microbial yield or the ratio of acetate to propionate. The ratio between MRT of the particulate and the liquid phase was smaller for sheep than that for cattle, and was not affected by treatment. Differences in this ratio might explain why species reacted differently to the saliva-inducing agent, which might help to explain the discrepancy between species in the effect of induced saliva flow on digestive parameters.

Macrowear effects of external quartz abrasives of different size and concentration in rabbits (Oryctolagus cuniculus).

External quartz abrasives are one of the driving forces of macrowear in herbivorous animals. We tested to what extent different sizes and concentrations influence their effect on tooth wear. We fed seven pelleted diets varying only in quartz concentration (0%, 4%, and 8%) and size (fine silt: ∼4 µm, coarse silt: ∼50 µm, fine sand: ∼130 µm) to rabbits (Oryctolagus cuniculus, n = 16) for 2 weeks each in a randomized serial experiment. Measurements to quantify wear and growth of incisors and the mandibular first cheek tooth, as well as heights of all other cheek teeth, were performed using calipers, endoscopic examination, and computed tomography scans before and after each feeding period. Tooth growth showed a compensatory correlation with wear. Absolute tooth height (ATH) and relative tooth height (RTH); relative to the 0% quartz "control" diet) was generally lower on the higher concentration and the larger size of abrasives. The effect was more pronounced on the maxillary teeth, on specific tooth positions and the right jaw side. When offered the choice between different sizes of abrasives, the rabbits favored the silt diets over the control and the fine sand diet; in a second choice experiment with different diets, they selected a pelleted diet with coarse-grained sand, however. This study confirms the dose- and size-dependent wear effects of external abrasives, and that hypselodont teeth show compensatory growth. The avoidance of wear did not seem a priority for animals with hypselodont teeth, since the rabbits did not avoid diets inducing a certain degree of wear.

Reticular contraction frequency and ruminal gas dome development in goats do not differ between grass and browse diets.

In investigations of differences between ruminant species feeding on browse or grass, it is often unclear whether observed differences are animal- or forage-specific. Ruminant species have been classified as 'moose-type', with little rumen content stratification, or 'cattle-type' with a distinct rumen contents stratification, including a gas layer. To which extent putative differences in forestomach motility are involved in these patterns is unknown. Using sonography, we investigated the frequency of reticular contractions and the stratification of rumen contents in goats fed exclusively on grass hay (n = 6) or dried browse (n = 5) directly after feeding, and after another 6 and 12 h with no access to feed. The frequency of reticular contractions decreased from immediately after feeding (1.8 ± 0.3 min-1 ) to 6 h afterwards (1.2 ± 0.2 min-1 ) and then remained constant, with no difference between diets. A gas dome became more visible over time, but neither its incidence nor its extent differed between diets. The results are in accord with classifying goats as 'cattle-type' in terms of their digestive physiology, and they add to a growing body of evidence that differences in digestive physiology between ruminant species are more due to species characteristics than different kinds of ingested forages.

Tooth wear, growth and height in rabbits (Oryctolagus cuniculus) fed pelleted or extruded diets with or without added abrasives.

Among the different factors thought to affect dental wear, dietary consistency is possibly the least investigated. To understand tooth wear of herbivorous animals consuming different dietary consistencies with different abrasive potential, we fed 14 rabbits (Oryctolagus cuniculus) exclusively with a timothy grassmeal-based diet in either pelleted or extruded form, or the same diets with an addition of 5% fine sand abrasives (mean size 130 µm). First, we offered the rabbits the pelleted and extruded diets as well as the pelleted control and pelleted abrasive diet in a two-stage preference experiment. Then, the rabbits received each diet for 2 weeks in a randomised serial feeding experiment, where each animal served as its own control. Tooth measurements for wear, growth and height were achieved using a manual calliper, endoscopic examination and CT scans. The analysis of the diets as fed showed almost identical mean particle size, but the extruded diet had a lower density (volume/mass) and softer consistency compared to the pelleted one and was favoured by most rabbits. The rabbits selected against the diet with sand during the preference experiment, possibly because it caused more tooth wear, especially on the teeth most exposed to wear along the upper tooth row (upper P4 and M1). The maxillary teeth also showed evidence of an increased chewing laterality by the end of the experiment. The extruded diet led to a significantly lower cheek teeth height than the pelleted diet, potentially due to the higher chewing effort needed for a similar dry matter intake. The results suggest that dietary hardness alone is a poor predictor of dental wear. The regrowth of the teeth matched wear consistently.

Increasing feed intake in domestic goats (Capra hircus): Measured effects on chewing intensity are probably driven by escape of few, large particles from the forestomach.

On the one hand, oral processing - mastication - is considered a relatively inflexible component of mammalian feed acquisition that constrains instantaneous intake rates. On the other hand, experimental data shows that the level of feed intake affects faecal particle size and hence net chewing efficiency in ruminants, with larger particles occurring in the faeces at higher intakes. Here, we report the effect of an increased feed intake during maintenance (L1), late (200% of L1) and peak lactation (300% of L1) of a consistent diet (hay:concentrates 50:50) in eight domestic goats on various measures of digestive physiology including faecal mean particle size (MPS). Increasing intake led to an increased gut fill, a reduction in digesta retention times, and an increase in faecal MPS (from 0.57 to 0.72 mm). However, this was an effect of the large particle fraction (>2 mm) being disproportionately excreted at higher intakes; if MPS was assessed on the basis of particles below the typical escape threshold (≤1 mm), there was no difference between intake levels. These findings suggest that the effect of intake on the calculated net chewing efficiency in ruminants may rather be an effect of increased large particle escape from the forestomach than a reduced chewing intensity per bolus during ingestion or rumination.

Dietary phosphorus restriction affects bone metabolism, vitamin D metabolism and rumen fermentation traits in sheep.

Homeostasis of calcium (Ca) and phosphate (Pi ) is maintained by a concerted interplay of absorption and reabsorption via the gastrointestinal tract and the kidney and by storage and mobilization from the bone regulated mainly by parathyroid hormone (PTH), 1,25-dihydroxycholecalciferol and calcitonin. The present study aimed at characterizing the effects of dietary P restriction on bone, vitamin D metabolism and rumen fermentation traits reflecting the endogenous P cycle maintaining the ruminal P supply for microbial metabolism. The experiments were done in eleven female, non-pregnant, non-lactating four- to nine-year-old Black Headed Mutton sheep allotted to two feeding groups: "P-restricted" (0.11% P/kg DM and 0.88% Ca/kg DM) and "Control" (0.38% P/kg DM and 0.88% Ca/kg DM). Dietary P restriction did not lead to hypophosphataemia, probably due to a compensation by bone mobilization, demonstrated by increased serum concentrations of a resorption marker and altered gene expression in bone tissue. In addition, the RNA expression of fibroblast growth factor 23, a bone-derived factor involved in the regulation of vitamin D metabolism, was significantly reduced with dietary P restriction. Furthermore, several genes related to vitamin D metabolism and plasma concentrations of 1,25-(OH)2 D were associated with serum concentrations of phosphate (Pi ). In the parotid gland, the expression of the Pi transporter NaPi2b was negatively associated with serum Pi and positively with parathyroid PTH expression. Although Pi concentrations in saliva and the gastrointestinal tract were significantly reduced, we found no adverse effects of the P-restricted ration on the production of short chain fatty acids, but slight differences in the production of butyrate as well as its relationship to rumen Pi and ammonia concentrations that might indicate an impact on ruminal fermentation.

Preliminary evidence for a forestomach washing mechanism in llamas (Lama glama).

Dust and grit are ingested by herbivores in their natural habitats along with the plants that represent their selected diet. Among the functions of the rumen, a washing of ingesta from adhering dust and grit has recently been demonstrated. The putative consequence is a less strenuous wear on ruminant teeth by external abrasives during rumination. The same function should theoretically apply to camelids, but has not been investigated so far. We fed six llamas (Lama glama) a diet of grass hay and a lucerne-based pelleted food in which fine sand had been included at about 8% of ingredients, for ad libitum consumption for 6 weeks. Subsequently, animals were slaughtered and content of the different sections of the gastrointestinal tract was sampled for the analysis of dry matter (DM), total ash, and acid detergent insoluble ash (ADIA, a measure for silica). Additionally, two of the animals were subjected to whole-body computer tomography (CT) after death in the natural sternal resting position. No clinical problems or macroscopic changes in the faeces were observed during the experimental period. The results indicate an accumulation of ADIA in the C3 compartment of the stomach complex, in particular in the posterior portion that is the equivalent of the abomasum in ruminants. By contrast, contents of the C1, from which material is recruited for regurgitation and rumination, were depleted of ADIA, indicating that the contents had largely been washed free of sand. The washing effect is an unavoidable side effect of the flotation- and sedimentation-based sorting mechanisms in the ruminant and the camelid forestomachs. In theory, this should allow ruminants and camelids to live in similar habitats as nonruminant herbivores at lower degrees of hypsodonty.

Validation of the RumiWatch Converter V0.7.4.5 classification accuracy for the automatic monitoring of behavioural characteristics in dairy cows.

The objective of the present study was to validate the accuracy of algorithms, implemented in the currently available RumiWatch Converter (RWC) version V0.7.4.5 of the RumiWatch System (RWS), for the classification of behavioural characteristics from jaw and head movements which are monitored by a noseband halter comprising a pressure sensor and a triaxial accelerometer. The accurate classification of behavioural characteristics in different time resolutions is critical for the usage of the RWS for scientific and practical purposes as chewing behaviour provides essential indicators for the assessment of diet adequacy in dairy cows. To validate the RWC V0.7.4.5 classification accuracy for behavioural characteristics of rumination, eating, drinking, other activity and ruminating chews per bolus by direct observation as reference method, 14 dairy cows participated in the trial. Concordance between the consolidated 1-min and 1-h classification results was assessed. The RWC V0.7.4.5 classified only rumination and ruminating chews per bolus precisely, whereas an algorithm optimisation for the classification of eating, drinking and other activity is required. Additionally, classification results from the 1-min and 1-h time summaries were not in agreement with each other except for rumination.

Digesta passage in nondomestic ruminants: Separation mechanisms in 'moose-type' and 'cattle-type' species, and seemingly atypical browsers.

Ruminants have been classified as having a 'moose-type' or 'cattle-type' digestive physiology. 'Cattle-type' ruminants have a clear difference in the mean retention time (MRT) of fluid vs. small particles in the reticulorumen (RR), with a high 'selectivity factor' (SF = MRTparticle/ MRTfluid, >1.80), and are typically grazers and intermediate feeders. 'Moose-type' ruminants have lower SF (<1.80), possibly because of defensive salivary proteins that constrain amounts of (high-viscosity) saliva, and are typically restricted to browsing. To further contribute to testing this physiology-diet correlation, we performed 55 individual passage measurements in 4/6 species that have/have not been investigated previously, respectively. Co-EDTA was used as a solute (fluid) and Cr-mordanted hay particles (<2 mm) as particle markers. Results are related to the percentage of grass in the natural diet taken from the literature. Moose (Alces alces, n = 4 on 4 to 5 diets each and n = 2 on a single diet, 5% grass, SF 1.46 ±â€¯0.22) and giraffe (Giraffa camelopardalis, n = 3 on 3 to 5 diets each, 1%, 1.42 ±â€¯0.23) as classical 'moose-type', and cattle (Bos taurus, n = 2, 70%, 2.04) as classical 'cattle-type' ruminants yielded results similar to those previously published, as did waterbuck (Kobus ellipsiprymnus, n = 5, 84%, 2.46 ±â€¯0.49), corroborating that the SF represents, to a large extent, a species-specific characteristic. Results in oryx (Oryx leucoryx, n = 1, 75%, 2.60) and sitatunga (Tragelaphus spekii, n = 4, 68%, 1.81 ±â€¯0.21) correspond to the concept of 'cattle-type' ruminants being grazers or intermediate feeders. However, European bison (Bison bonasus, n = 1, 10%, 2.74), nyala (T. angasii, n = 6, 20%, 1.95 ±â€¯0.25), bongo (T. eurycerus, n = 3, 13%, 2.39 ±â€¯0.54) and gerenuk (Litocranius walleri, n = 1, 0%, 2.25) appear as 'cattle-type' ruminants, yet have a browse-dominated diet. While the results do not challenge the view that a 'moose-type' digestive physiology is an adaptation to browse diets, they indicate that it may not be the only adaptation that enables ruminants to use browse. Apparently, a 'cattle-type' digestive physiology with a high SF does not necessarily preclude a browsing diet niche. High-SF browsers might have the benefit of an increased harvest of RR microbiota and grit removal prior to rumination; how they defend themselves against secondary plant compounds in browse remains to be investigated.

Microbial protein formation of different carbohydrates in vitro.

The aim of this study was to investigate the microbial protein yield of different pure carbohydrates to contribute to a more precise prediction of the microbial protein formed in the rumen. In a first experiment, sucrose, wheat starch, microcrystalline cellulose and citrus pectin were incubated for 8 and 24 hr in the modified Hohenheim gas test (HGT) system (3 runs × 2 syringes) including gas production, ammonia and short-chain fatty acid concentration measurements. Ammonia values were used for estimation of the microbial protein formation. In a second experiment, the same substrates were incubated for 96 hr in the HGT system (2 runs × 3 syringes) and gas production was measured after 2, 4, 6, 8, 12, 16, 24, 30, 36, 48, 60, 72 and 96 hr of incubation to obtain the fermentation kinetics and the time of half-maximal gas production (t1/2 ) of the substrates. The substrates differed considerably in their fermentation kinetics, and therefore, comparison on the basis of t1/2 was chosen as the most meaningful. At t1/2 , microbial protein yield [g/kg dry matter] was higher for cellulose than for sucrose and pectin and higher for starch than for sucrose. The microbial protein expressed in g/L gas production was higher for starch and cellulose than for sucrose and pectin at t1/2 . Effects of carbohydrates related to ruminal pH may remain undetected in in vitro trials.

Contribution of different rumen microbial groups to gas, short-chain fatty acid and ammonium production from different diets-an approach in an in vitro fermentation system.

In this study, the relative contribution of different microbial groups to ruminal metabolism was investigated for different diets. The rumen microbial cultures included whole rumen fluid, fungi + protozoa, bacteria + protozoa, protozoa and bacteria + fungi and were established by physical and chemical methods. Gas production, short-chain fatty acid (SCFA) and ammonium production were measured at 24 hr in in vitro incubations using the Hohenheim gas test (HGT) procedure. Seven donor animal diets with different concentrate-to-roughage ratios (C:R: 10:90, 30:70, 50:50, 70:30, 70:30BC (BC = NaHCO3 ), 90:10 and 90:10BC) and five HGT diets (C:R: 10:90, 30:70, 50:50, 70:30 and 90:10) were formulated. Incubations in the HGT were always based on inoculum from sheep diets with the respective C:R ratio. Gas and ammonium production increased (p < 0.001) as a result of a gradual increase in concentrate proportion of the diets. In general, SCFA production followed the same trend. Whole rumen fluid and bacteria + fungi produced approximately 50% higher gas volume than protozoa and fungi + protozoa fractions, whereas gas production with bacteria + protozoa was at an intermediate level. Coculture of protozoa either with bacteria or with fungi produced more ammonium. Populations without bacteria were characterized by a particularly high acetate/propionate ratio. Although an interaction between microbial group and diet was observed for several variables, no clear direction could be established. Manipulating rumen fluid by selectively suppressing specific rumen microbial groups may be a helpful tool in elucidating their role in nutrient degradation and turnover in vitro.

Controlled feeding experiments with diets of different abrasiveness reveal slow development of mesowear signal in goats (Capra aegagrus hircus).

Dental mesowear is applied as a proxy to determine the general diet of mammalian herbivores based on tooth-cusp shape and occlusal relief. Low, blunt cusps are considered typical of grazers and high, sharp cusps typical of browsers. However, how internal or external abrasives impact mesowear, and the time frame the wear signature takes to develop, still need to be explored. Four different pelleted diets of increasing abrasiveness (lucerne, grass, grass and rice husks, and grass, rice husks and sand) were fed to four groups of a total of 28 adult goats in a controlled feeding experiment over a 6-month period. Tooth morphology was captured by medical CT scans at the beginning and end of the experiment. These scans, as well as the crania obtained post mortem, were scored using the mesowear method. Comparisons between diet groups showed few significant differences after 6 months, irrespective of whether CT scans or the real teeth were scored. Only when assessing the difference in signal between the beginning and the end of the experiment did relevant, significant diet-specific effects emerge. Diets containing lower phytolith content caused a more pronounced change in mesowear towards sharper cusps/higher reliefs, while the feed containing sand did not result in more extreme changes in mesowear when compared with the same feed without sand. Our experiment suggests that the formation of a stable and hence reliable mesowear signal requires more time to develop than 6 months.

Decreasing methane yield with increasing food intake keeps daily methane emissions constant in two foregut fermenting marsupials, the western grey kangaroo and red kangaroo.

Fundamental differences in methane (CH4) production between macropods (kangaroos) and ruminants have been suggested and linked to differences in the composition of the forestomach microbiome. Using six western grey kangaroos (Macropus fuliginosus) and four red kangaroos (Macropus rufus), we measured daily absolute CH4 production in vivo as well as CH4 yield (CH4 per unit of intake of dry matter, gross energy or digestible fibre) by open-circuit respirometry. Two food intake levels were tested using a chopped lucerne hay (alfalfa) diet. Body mass-specific absolute CH4 production resembled values previously reported in wallabies and non-ruminant herbivores such as horses, and did not differ with food intake level, although there was no concomitant proportionate decrease in fibre digestibility with higher food intake. In contrast, CH4 yield decreased with increasing intake, and was intermediate between values reported for ruminants and non-ruminant herbivores. These results correspond to those in ruminants and other non-ruminant species where increased intake (and hence a shorter digesta retention in the gut) leads to a lower CH4 yield. We hypothesize that rather than harbouring a fundamentally different microbiome in their foregut, the microbiome of macropods is in a particular metabolic state more tuned towards growth (i.e. biomass production) rather than CH4 production. This is due to the short digesta retention time in macropods and the known distinct 'digesta washing' in the gut of macropods, where fluids move faster than particles and hence most likely wash out microbes from the forestomach. Although our data suggest that kangaroos only produce about 27% of the body mass-specific volume of CH4 of ruminants, it remains to be modelled with species-specific growth rates and production conditions whether or not significantly lower CH4 amounts are emitted per kg of meat in kangaroo than in beef or mutton production.