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.     

Effects of heat treatment on protein feeds evaluated in vitro by the method of estimating utilisable crude protein at the duodenum

The effect of heat treatment on the protein value of field beans, lupins and peas was studied using an in vitro method. Protein feeds were subjected to heat treatment for 30, 60 and 90 min in forced air oven at 120, 140 and 160 °C and in autoclave at 105, 120 and 135 °C. The heat‐treated protein feeds were incubated in buffered rumen fluid together with grass silage and barley in complete isonitrogenous diets. The gas production (GP) was recorded continuously, and ammonia‐N (NH₃‐N) concentrations were determined during the in vitro incubation at 8, 24 and 48 h and used to determine the utilisable crude protein (uCP) at the duodenum at 16 h of incubation (uCP₁₆). Heat treatments decreased the concentration of soluble crude protein and increased neutral detergent insoluble CP (NDICP) in all protein feeds compared to untreated. Inclusion of protein feeds to basal diet showed no increase in the uCP₁₆ in untreated field bean diet and only a small numerical increase in the uCP₁₆ concentrations from 160 g/kg dry matter (DM) to 166 and 172 g/kg DM in untreated lupine and pea diets, respectively, indicating high degradability of untreated feeds. Increasing the time and temperature of the heat treatment linearly increased the uCP₁₆ concentrations in field bean and pea diets, but not in lupin diets. Autoclave treatment was more effective in decreasing uCP₁₆ than oven treatment despite the lower temperatures used. However, the combination of highest temperatures and treatment time in autoclave increased acid detergent insoluble CP (ADICP) concentrations in protein feeds, indicating protein damage and decreased intestinal digestibility. Determining in vitro uCP and ADICP shows to be a promising method for evaluating protein value in heat‐treated animal feeds.     

Influence of foetal calf serum supplementation during in vitro embryo culture in Iberian red deer

Nowadays, the use of foetal calf serum (FCS) during in vitro embryo culture is very controversial. Whilst some authors have encouraged its use, others reject it because of its harmful effects. Although in vitro embryo production in red deer is a promising assisted reproductive technique, it is still in its infancy and a great effort is needed to update the protocols used. The aim of this study was to assess whether FCS supplementation in red deer embryo culture medium is necessary to produce blastocyst and, if so, when is the best time to add it in terms of blastocyst production and quality. In vitro blastocysts were cultured with FCS added at 24, 48 or 96 hours post‐insemination (hpi). In addition, a treatment without FCS was used as control. Six hundred and ninety‐four cumulus–oocyte complexes were collected for in vitro fertilization. Cleavage rate was examined at 48 hpi, and blastocyst yield was recorded on days 6, 7 and 8. FCS had no influence on cleavage and blastocyst rate for any of the treatments studied. However, the number of cells was higher (p = .025) in those blastocysts cultured with FCS from 48 hpi compared with FCS‐free culture media (93.88 ± 7.76 vs. 54.11 ± 8.36). In conclusion, the addition of FCS to the embryo culture medium at 48 hpi improves the quality of red deer blastocyst, although it does not affect the percentage of embryos obtained.     

Seroprevalence of Toxoplasma gondii in wild boar and deer in Brandenburg,Germany

Consumption of game in Germany has increased during the past 10 years. Wild boar (Sus scrofa), roe deer (Capreolus capreolus) and red deer (Cervus elaphus) are the most frequently hunted and consumed game animals in Germany, yet information on the occurrence of zoonotic pathogens in these animal species is scarce. To better estimate the public health risk emanating from handling and consumption of game, this study investigated seroprevalences of Toxoplasma gondii in game hunted in the German federal state of Brandenburg during two hunting seasons from 2017 to 2019. Toxoplasma gondii‐specific antibodies were detected in 24.4% (44/180, 95% CI: 18.4%–31.4%) of wild boar, 12.8% (16/125, 95% CI: 7.5%–20%) of roe deer and 6.4% (3/47, 95% CI: 1.3%–17.5%) of red deer using a commercial ELISA kit. Seroprevalences were similar in the two hunting seasons. Correlation between sex and seropositivity could not be observed. A rise in seroprevalence was seen with increasing age in all studied game species. Observed seroprevalences suggest that T. gondii is endemic in the sylvatic environment in the German federal state of Brandenburg and imply that game could represent a relevant source for human T. gondii infection.     

Red urine from red deer grazed on pure red clover swards

Twenty-four red deer hinds with their calves were released on to a newly established pure red clover sward and, 2 days later, red staining of the tail, perineum and hocks was observed. This was presumed to be of urinary origin. Observation of micturition showed that when urine was passed, it was a normal straw colour but it turned scarlet-red about 1 hour after exposure to air. Midstream urine remained the normal colour when held under a pure nitrogen atmosphere immediately after micturition, but it turned red when held in air in the dark, suggesting that the colour change was due to an oxidative rather than a photosensitive reaction. All deer grazing red clover were affected but this did not occur in deer grazing ryegrass/white clover swards. No adverse effects were observed in the deer grazing the red clover, and calf growth was significantly higher than on ryegrass/white clover, suggesting that the red urine had no effect on health or productivity.

Blood and urine analyses showed no signs of haemolysis, haematuria or haemoglobinuria. Preliminary chemical analyses suggest that the compounds involved are not those found in the urine of sheep grazing oestrogenic clover. The nature of the compounds have yet to be determined.     

Diurnal Patterns of Ruminal Concentrations and Portal Appearance Rates of Short-chain Fatty Acids in Sheep Fed a Hay or a Concentrate/Straw Diet in Two Meals Daily

Abstract

Three rumen fistulated and catheterized sheep were meal-fed and used to study ruminal and arterial concentrations of short-chain fatty acids (SCFA) as well as portal appearance rates of SCFA and irreversible loss rate (ILR) of acetate in 24 h periods on a hay and a concentrate/straw diet, respectively.

Ruminal and arterial concentrations as well as portal appearance rates of SCFA and ILR of acetate were significantly affected by the intake of feed. Generally, the highest concentrations and appearance rates were obtained 2 h after feeding. The portal recovery of arterial acetate was not affected by feeding or diet. The 24 h means were 0.68 ± 0.01 and 0.67 ± 0.01 on the hay and the concentrate/straw diet, respectively. Partial correlation coefficients corrected for the effects of time, sheep, and diet were calculated for the relationships evaluated. The portal appearance rate of acetate (r = 0.52, P <0.001) and the portal net appearance rate of propionate (r = 0.68, P <0.001) were linearly related to the ruminal concentrations of the two SCFA. The logarithm of the portal net appearance rate of butyrate seemed to be linearly related to the logarithm of the ruminal concentration of undissociated butyric acid (r = 0.70, P <0.001) when the effect of time was omitted from the model. The portal appearance rate of acetate (r = 0.22, P <0.05) and the portal net appearance rate of propionate (r = 0.63, P <0.001) as well as butyrate (r = 0.55, P <0.001) were linearly related to the arterial concentration of the respective SCFA. The results show that within animal and diet the ruminal as well as arterial concentrations are good predictors of SCFA portal appearance rates in sheep fed roughage at maintenance. Ruminal and arterial concentrations of SCFA seem less reliable predictors of portal appearance rates of SCFA between diets and sheep. The portal appearance of SCFA, 5.6 ± 0.5 and 7.0 ± 0.3 mol d^?1, accounted for 44 ± 4 and 43 ± 2% of the calculated metabolizable energy intake on the hay and the concentrate/straw diets, respectively.     

Ontogenesis of the rumen: A comparative analysis of the Merino sheep and Iberian red deer

The aim of this study is to describe differences in the ontogenesis of the rumen in the sheep (domestic ruminant) and deer (wild ruminant). A total of 50 embryos and fetuses of Merino sheep and 50 of Iberian deer were used, from the first stages of prenatal life until birth. For the study, the animals were divided into five experimental groups according to the most relevant histological characteristics. The appearance of the rumen from the primitive gastric tube was earlier in the sheep (22% gestation, 33 days) than in the deer (25% gestation, 66 days). In both cases it displayed a primitive epithelium of a stratified, cylindrical, non‐ciliary type. At around 28% gestation in the sheep (42 days) and 26% (67 days) in the deer, the rumen was configured of three clearly‐differentiated layers: internal or mucosal, middle or muscular and external or serosal. In both species the stratification of the epithelial layer was accompanied by modifications in its structure with the appearance of the ruminal pillars and papillae. The pillars appeared before the papillae and the appearance of both structures was always earlier in the deer (pillars: 70 days, 27% gestation; papillae: 97 days, 36% gestation) than in the sheep (pillars: 42 days, 28% gestation; papillae: 57 days, 38% gestation). The outlines of the ruminal papillae appeared as evaginations of the basal zone toward the ruminal lumen, dragging in their formation the basal membrane, the lamina propria and the submucosa. The tegumentary mucosa of the rumen was without secretion capability in the first embryonic phases. From 67 days (26% gestation) the neutral mucopolysaccharides appeared in the deer and at 46 days (30% gestation) in the sheep. In both cases they continued to decrease until birth, this diminution being more pronounced in the deer. Finally, the presence of neuroendocrine and glial cells was detected in the deer at earlier stages than in the sheep.     

Influence of calf genotype on dam lactation and calf growth in farmed red deer (Cervus elaphus)

Multiparous red deer (Cervus elaphus scoticus) hinds (n=17) were either mated to a red deer stag or artificially inseminated with semen from a wapiti bull (C.e. nelsoni) to produce red deer or F₁ crossbred (C.e. scoticus × C.e. nelsoni) calves to test the hypotheses that (1) red deer hinds rearing crossbred calves will produce significantly more milk to support calf growth than hinds rearing red deer calves, and (2) hind live weight and body condition may suffer as a result. Hinds and calves were grazed on ryegrass and white clover pastures and supplemented with pasture silage and barley grain when pasture supply was inadequate. Calves were left with their mothers until 9–10 months of age when lactation had terminated naturally. Mean body condition score (BCS) loss was greater in hinds rearing F₁ calves than in hinds rearing red calves in the period from late gestation to mid-lactation (September–March, P<0.05). F₁ calves grew significantly faster than red deer calves and were heavier at all ages. From February onwards there was a consistent decline in milk intake, and by the latest measurement (22 July) milk production was negligible, and most hinds had ceased lactating. Milk intake by the F₁ calves was significantly greater until day 114, after which the measured milk intake was not significantly different. Pasture intake of the F₁ calves was significantly greater at day 114 but not at other times (P<0.05). The average hind pasture intake was also not significantly affected by calf genotype when adjusted for post calving hind live weight. The results support the hypothesis that a red deer hind rearing an F₁ calf produces significantly more milk than a hind rearing a red deer calf, suggesting that the growth of a red deer calf is limited by its milk demand rather than milk supply by the hind. The study also supported the second hypothesis. Hinds rearing F₁ calves exhibited greater losses of body condition score relative to the hinds rearing red deer calves, indicating that the hinds rearing F₁ calves were in a greater negative energy balance that hinds rearing red deer calves. This highlights the additional energetic demands on a hind from feeding an F₁ calf and in a farmed context the need to ensure that adequate nutrition is provided.     

Intake and utilization of sorghum and maize green fodder in spotted deer under captivity

Purpose  

The feeding value and nutritional adequacy of two cereal fodder, viz. sorghum (Sorghum vulgare) and maize (Zea mays), were evaluated in spotted deer (Axis axis) and compared with that in sheep in two consecutive feeding trials of 35 days each.     

Effect of tropical browse leaves supplementation on rumen enzymes of sheep and goats fed <Emphasis Type="Italic">Dichanthium annulatum</Emphasis> grass-based diets

In a switch-over experiment, eight male animals, four each of sheep and goats of local breeds with mean body weight of 26. 8 ± 2.0 and 30.0 ± 2.1 kg, were fed Dichanthium annulatum (DA) grass and four browse species viz. Helictris isora, Securengia virosa, Leucaena leucocephala (LL) and Hardwickia binnata (HB) in four feeding trials to assess their supplementary effect on activity of rumen enzymes. The sheep and goats were offered DA grass with individual browse in 75:25 and 50:50 proportions, respectively, for more than 3 months during each feeding trial, and rumen liquor samples were collected twice at 0 and 4 h post feeding after 60 and 90 days of feeding. Glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and glutamate dehydrogenase (GDH) enzymes were determined in the bacteria and protozoa fractions of rumen liquor, while cellulase enzyme activity was measured in mixed rumen liquor. LL and HB had the highest and lowest contents of CP, while fibre contents were lower in early than later browse leaves. Supplementation of browse leaves significantly (P < 0.05) affect the specific activity of GDH enzyme in bacteria fraction of rumen liquor of animal species, while GDH activity was similar in protozoa fraction of rumen liquor of sheep and goats on all DA grass–browse-supplemented diets except DA–HB (42.8 units/mg protein), where activity was significantly (P < 0.05) low. Specific activities of GOT and GPT enzymes in both bacteria and protozoa fractions of rumen liquor differ significantly (P < 0.05) due to supplementation of browse leaves to DA grass. Browse leaves significantly (P < 0.05) affect the cellulase enzyme activity in animal rumen liquor, being highest on DA–LL (193.4) and lowest on DA–HB diet (144.8 μg sugar/mg protein). Goat exhibited higher activities of GOT and GPT than sheep in both bacteria and protozoa fraction of rumen liquor, while cellulase activity was similar between the animal species on the grass–browse leaves diets. Results indicate that browse leaves supplementation affect the enzyme activities of sheep and goats rumen, while the goats rumen liquor had higher activities of GOT, GPT and GDH enzyme than sheep.     

Experimental tuberculosis in red deer (Cervus elaphus)

This study was designed to investigate experimental Mycobacterium bovis infection of red deer (Cervus elaphus). Three intravenously inoculated deer (dose 10µg–1000µg) developed miliary tuberculosis of the lungs and all died within 28 days of being infected. No clinical illnesses were observed in four subcutaneously (dose 1 uµg–100uµg) and three intratracheally (dose lµg–100µg) inoculated deer. At the conclusion of the experiment six weeks post inoculation, these seven animals reacted to 2 mg/ml of bovine purified protein derivative. The principal lesions in the intravenously inoculated deer were in the lungs which had multiple foci of necrosis containing very large numbers of acid fast bacilli. A gradation of changes was seen in the subcutaneously inoculated deer. The animal receiving the 1µg dose only had lesions at the injection site and the draining prescapular lymph node. Deer receiving higher doses also had histopathological changes in the lungs and liver. Microscopic changes in the intratracheally infected animals were restricted to the thoracic cavity. The ability of the deer to control infection was related to the route of inoculation.     

Leptospiral antibodies in Iberian red deer (Cervus elaphus hispanicus), fallow deer (Dama dama) and European wild boar (Sus scrofa) in Asturias,Northern Spain

Serum samples collected from Iberian red deer (Cervus elaphus hispanicus; n = 472), fallow deer (Dama dama; n = 293) and European wild boar (Sus scrofa; n = 174) in Asturias, Northern Spain, from 1999 to 2005 were examined for antibodies against a reference panel of 14 Leptospira spp. serovars. Positive antibody titres at a microscopic agglutination test cut-off of 1:80 were detected against serovars Pomona (1.6%, 5.8%, 5.2%), Bratislava (1.1%, 0.7%, 4.7%), Grippotyphosa (0.7%, 2.4%, 1.7%), Muenchen (2.6%, 0%, 0%), Pyrogenes (0.4%, 2.4%, 1.2%), Panama (1.2%, 1.7%, 0%), Copenhageni (0%, 0.7%, 0.6%), Autumnalis (0.4%, 0%, 0.6%) and Icterohaemorrhagiae (0%, 0%, 0.6%) in Iberian red deer, fallow deer and European wild boar, respectively.     

Comparative analysis of the merino sheep and Iberian red deer abomasum during prenatal development

The aim of this study is to describe differences in the ontogenesis of the abomasum in sheep (domestic ruminant) and deer (wild ruminant). Histomorphometric and immunohistochemical analysis were carried out on 50 embryos and fetuses of the sheep and 50 red deer from the first prenatal stages until birth. To compare similar periods of gestation in both species, we calculate the percentages of gestation. The appearance of the abomasum was earlier in the red deer (22% gestation) than in the sheep (25% gestation). Throughout development the epithelium happened sequentially, being of the types pseudostratified to simple cylindrical. This important modification was earlier in the red deer than the sheep. At 46% gestation in red deer and 50% in sheep, gastric pits were observed on the surface of abomasal folds. Our studies suggest a close link between the initial formation of these pseudoglandular structures and the clear separation of lamina propria and submucosa separated by de muscularis mucosae. At 54% gestation in red deer and at 60% in sheep, in the bottom of these pits the first outlines of glands were distinguishable. Finally, the presence of neuroendocrine and glial cells were detected in deer at earlier stages than in sheep.     

Keratitis in red deer

Twenty-five of 40 red deer (Cervus elaphus) exhibited corneal opacities and ulceration resembling mild bovine infectious keratitis. Attempts to incriminate either a bacterial or viral causal agent were unsuccessful.

Subconjunctival injections of a penicillin/streptomycin mixture were administered in both eyes of affected and in-contact animals. Resolution over an 8 week period was uneventful.     

Comparison of concentrations of copper in plasma and serum from farmed red deer (Cervus elaphus)

AIM: To determine the relationship between the concentrations of Cu in plasma and serum in red deer, and to compare this relationship with those previously reported in cattle and sheep.

METHODS: Paired serum and heparinised plasma samples from 114 red deer from 10 herds (n=6-20 per herd) were analysed for concentrations of Cu. Samples were collected either at slaughter (n=84; eight herds) or by jugular venepuncture (n=30; two herds). Thirty-nine of the samples taken at slaughter were from adult hinds from four herds, while other samples were taken from 10–14-month-old males, except for one herd (10 samples) where an equal number of 8–9-month-old males and females were sampled. The effect of age, gender and herd on the relationship between concentrations of Cu in plasma and serum was assessed using univariate ANOVA. The individual results for concentrations of Cu in serum were compared with those in plasma, using limits-of-agreement plotting.

RESULTS: The mean concentration of Cu in plasma was not significantly different from that of serum (0.048; 95% CI=-0.14 to 0.24 µmol/L). There was no effect of age, sex or herd on this relationship.

CONCLUSIONS: In deer, there was no significant difference between concentrations of Cu in plasma and serum regardless of age, sex or herd of origin.

CLINICAL RELEVANCE: In contrast to the situation in cattle and sheep, the concentration of Cu in serum can be used interchangeably with that in plasma for the estimation of concentration of Cu in blood of red deer.     

Animal,feed and rumen fermentation attributes associated with methane emissions from sheep fed brassica crops

Methane emissions from ruminants enhance global warming and lead to a loss of feed energy. The emissions are low when fed brassica crops, but the factors contributing to low emissions are unknown. A meta-analysis was conducted with individual animal data collected from seven experiments. In these experiments, methane emissions were measured using respiration chambers. Animal characteristics, feed chemical composition and rumen fermentation parameters were included for the analysis using multiple regression models. Feed intake level, animal live weight and age were animal factors that were weakly and negatively related to methane yield (g/dry matter intake). The duration in which sheep were fed brassica crops was a significant contributor in the model, suggesting that the effect on emissions diminishes with time. Among a range of feed chemical composition characters, acid detergent fibre and hot-water-soluble carbohydrate contributed significantly to the model, suggesting that both structural and soluble carbohydrates affect methane formation in the rumen. There was no significant correlation between the concentration of sulphate in brassicas and emissions, but nitrate was moderately and negatively correlated with methane yield (r = −.53). Short-chain fatty acid profiles in the rumen of animals fed brassicas were different from those fed pasture, but these parameters only moderately correlated to methane emissions (r = .42). Feeding forage rape resulted in low rumen pH. The pH before morning feeding was strongly correlated to methane yield (r = .90). Rumen pH, together with microbial communities mediated by pH, might lead to low emissions. Bacteria known to produce hydrogen were relatively less abundant in the rumen contents of brassica-fed animals than pasture-fed animals. In conclusion, animal and feed factors, rumen fermentation and microbial communities all affect methane emissions to some extent. The interactions of these factors with each other thus contribute to methane emissions from brassica-fed sheep.     

Comparison of feed intake,digestion and rumen function among domestic ruminant species grazing in upland vegetation communities

This study aimed to compare feed intake, digestion, rumen fermentation parameters and bacterial community of 5 beef cows, 12 crossed ewes and 12 goats grazing together in spring–early summer on heather–gorse vegetation communities with an adjacent area of improved pasture. Organic matter intake (OMI) and digestibility (OMD) were estimated using alkane markers. Ruminal fluid samples were collected for measuring fermentation parameters, and studying the bacterial community using terminal restriction fragment length polymorphism (T‐RFLP). Spot samples of urine were taken to determine purine derivative (PD) and creatinine concentrations to estimate microbial protein synthesis in the rumen. Herbaceous species were the main dietary component in all animal species. Cattle had higher (p < 0.05) daily OMI (g/kg LW^0.75) and OMD, whereas sheep and goats showed similar values. The highest ammonia concentration was observed in sheep. Total VFA, acetate and butyrate concentrations were not influenced by animal species, while propionate concentrations in goats were 1.8 times lower (p < 0.05) than in sheep. Acetate:propionate ratio was greater (p < 0.05) in goats, whereas cattle excreted more allantoin (p < 0.05). Estimated supply of microbial N was higher in cows (p < 0.01), whereas the efficiency of microbial protein synthesis was lower (p < 0.01) in this animal species. Hierarchical clustering analysis indicated a clear effect of animal species on rumen bacterial structure. Differences among animal species were also observed in the relative frequency of several T‐RFs. Certain T‐RFs compatible with Lachnospiraceae, Proteobacteria and Clostridiales species were not found in goats, while these animals showed high relative frequencies of some fragments compatible with the Ruminococcaceae family that were not detected in sheep and cattle. Results suggest a close relationship between animals’ grazing behaviour and rumen bacterial structure and its function. Goats seem to show a greater specialization of their microbial populations to deal with the greater fibrous and tannin content of their diet.     

The supply of wild ruminants with major and trace elements. 2. The manganese content of winter grazing and the manganese status of red deer, fallow deer, roes and mouflons

The winter grazing of wild ruminants on mantles of slate waste in the Harz mountains and in Gera county were richest in Mn whereas those on mantles of granite waste in the Erzgebirge were poorest. The flora of the shell-limestone, keuper and loess areas contains much less Mn than that of those acid habitats. The Mn-requirement of the wild ruminants grazing in the forests is met however, since bilberry plants (2,080 mg/kg), spruce twigs (984 mg/kg), spruce bark (827 mg/kg), oak twigs (791 mg/kg) and heather (754 mg/kg dry matter) in addition to many other plant species store extremely high amounts of Mn. Solely sallow twigs were poor in Mn (28 mg/kg). Based on 601 samples examined, the Mn supply of wild ruminants is extensively described. The rumen content reflected the plentiful Mn-supply of the wild ruminants living in forests (greater than 400 mg/kg) and the far worse one of field roes, particularly in Mn-deficiency areas for domesticated ruminants (mantle of shell-limestone waste 37 mg Mn/kg dry matter of rumen content). The indicator organs of the Mn-status (liver, covering hair, kidneys) verify the statements made concerning red deer, fallow deer and mouflons, of which a total of 170 head from 14 biotopes were examined. An Mn-deficit of field roes in Mn-deficiency habitats in winter cannot completely be excluded. Mouflons have not yet been able to adapt themselves to the excessive Mn-supply of the acid forest habitats in Central Europe. They stored significantly higher amounts of Mn in liver, covering hair, kidneys, cerebrum and ribs than the other wild ruminants and sheep and cattle. The normal Mn-content of the liver and the cerebrum of red deer, fallow deer and roes corresponds to that of sheep and cattle. Roes and fallow deer have winter covering hair poor in Mn (less than 4.0 mg/kg) in comparison to sheep and cattle.     

Impact of variation in structure of condensed tannins from sainfoin (Onobrychis viciifolia) on in vitro ruminal methane production and fermentation characteristics

Our study investigated the effects of condensed tannins (CT) on rumen in vitro methane (CH₄) production and fermentation characteristics by incubating lucerne in buffered rumen fluid in combination with different CT extracts at 0 (control), 40, 80 and 120 g CT/kg of substrate DM. Condensed tannins were extracted from four sainfoin accessions: Rees ‘A’, CPI63763, Cotswold Common and CPI63767. Gas production (GP) was measured using a fully automated GP apparatus with CH₄ measured at distinct time points. Condensed tannins differed substantially in terms of polymer size and varied from 13 (Rees ‘A’) to 73 (CPI63767) mean degree of polymerization, but had relatively similar characteristics in terms of CT content, procyanidin: prodelphinidin (PC: PD) and cis:trans ratios. Compared to control, addition of CT from CPI63767 and CPI63763 at 80 and 120 g CT/kg of substrate DM reduced CH₄ by 43% and 65%, and by 23% and 57%, respectively, after 24‐h incubation. Similarly, CT from Rees ‘A’ and Cotswold Common reduced CH₄ by 26% and 46%, and by 28% and 46% respectively. Addition of increasing level of CT linearly reduced the maximum rates of GP and CH₄ production, and the estimated in vitro organic matter digestibility. There was a negative linear and quadratic (p < 0.01) relation between CT concentration and total volatile fatty acid (VFA) production. Inclusion of 80 and 120 g CT/kg of substrate DM reduced (p < 0.001) branched‐chain VFA production and acetate: propionate ratio and was lowest for CPI63767. A decrease in proteolytic activity as indirectly shown by a change in VFA composition favouring a shift towards propionate and reduction in branched‐chain VFA production varied with type of CT and was highest for CPI63767. In conclusion, these results suggest that tannin polymer size is an important factor affecting in vitro CH₄ production which may be linked to the CT interaction with dietary substrate or microbial cells.     

Red urine from red deer grazed on pure red clover swards

Twenty-four red deer hinds with their calves were released on to a newly established pure red clover sward and, 2 days later, red staining of the tail, perineum and hocks was observed. This was presumed to be of urinary origin. Observation of micturition showed that when urine was passed, it was a normal straw colour but it turned scarlet-red about 1 hour after exposure to air. Midstream urine remained the normal colour when held under a pure nitrogen atmosphere immediately after micturition, but it turned red when held in air in the dark, suggesting that the colour change was due to an oxidative rather than a photosensitive reaction. All deer grazing red clover were affected but this did not occur in deer grazing ryegrass/white clover swards. No adverse effects were observed in the deer grazing the red clover, and calf growth was significantly higher than on ryegrass/white clover, suggesting that the red urine had no effect on health or productivity. Blood and urine analyses showed no signs of haemolysis, haematuria or haemoglobinuria. Preliminary chemical analyses suggest that the compounds involved are not those found in the urine of sheep grazing oestrogenic clover. The nature of the compounds have yet to be determined.