Freezing African elephant semen as a new population management tool.

BACKGROUND: The captive elephant population is not self-sustaining and with a limited number of breeding bulls, its genetic diversity is in decline. One way to overcome this is to import young and healthy animals from the wild. We introduce here a more sustainable alternative method - importation of semen from wild bulls without removing them from their natural habitat. Due to the logistics involved, the only practical option would be to transport cryopreserved sperm. Despite some early reports on African elephant semen cryopreservation, the utility of this new population management tool has not been evaluated. METHODOLOGY/PRINCIPAL FINDINGS: Semen was collected by electroejaculation from 14 wild African savanna elephant (Loxodonta africana) bulls and cryopreserved using the directional freezing technique. Sperm treatments evaluated included the need for centrifugation, the use of hen or quail yolk, the concentration of glycerol (3%, 5% or 7%) in the extender, and maintenance of motility over time after thawing. Our results suggest that dilution in an extender containing hen yolk and 7% glycerol after centrifugation best preserved post-thaw sperm motility when compared to all other treatments (P≤0.012 for all). Using this approach we were able to achieve after thawing (mean ± SD) 54.6±3.9% motility, 85.3±2.4% acrosome integrity, and 86.8±4.6% normal morphology with no decrease in motility over 1 h incubation at 37°C. Sperm cryopreserved during this study has already lead to a pregnancy of a captive female elephant following artificial insemination. CONCLUSIONS/SIGNIFICANCE: With working techniques for artificial insemination and sperm cryopreservation of both African and Asian elephants in hand, population managers can now enrich captive or isolated wild elephant populations without removing valuable individuals from their natural habitat.

Variation in growth and potentially associated health status in Hermann's and spur-thighed tortoise (Testudo hermanni and Testudo graeca).

Captive reptiles often show higher growth rates than in the wild, possibly due to higher feeding intensity. Although health problems are usually linked to inappropriate diets, fast growth itself, such as triggered by appropriate diets fed in high amounts, has traditionally also been considered unfavorable for tortoises. We document growth rates (based on age and mass) from private Testudo hermanni and T. graeca breeders, which are generally higher than those reported for free-ranging specimens, but show enormous variation. Tortoise patients presented to an exotics clinic also covered the whole growth rate spectrum. To test whether fast growth was associated with diseases, the age-body mass relationship of these patients was tested, in a retrospective evaluation, for additional influence factors, such as dietary history and occurrence of certain diet and growth-related diseases. No indication was found that animals particularly heavy for their age were more prone to diet/growth-related disorders. In general, tortoises fed diets with meat/grain were heavier for their age than tortoises fed more appropriate diets; dietary history was not related to a particular disease. The results suggest the age-body mass relationship may not be suitable for testing effects of fast growth; an age-body length relationship would be more appropriate. Animals presented for a diet/growth-related disorder were younger than animals presented for other reasons; there was a significant negative correlation between the severity of pyramiding and age, suggesting that growth-related disorders may well limit the life expectancy of tortoises. Controlled clinical studies are required to fully test this hypothesis.

Condensing results of wet sieving analyses into a single data: a comparison of methods for particle size description.

Sieve analysis is used in feed analysis, and studies of digestive physiology with various approaches to describe an average value of particle size which can serve to compare different samples. To demonstrate the effects of such different approaches, we compared five particle size indicators to demonstrate advantages and disadvantages of each method, the modulus of fineness (MOF), the discrete mean (dMEAN) and median (dMED), and the continuous mean (cMEAN) and median (cMED), well aware of the fact that a gold standard for this procedure is lacking. Data were obtained from 580 individual faecal samples of different herbivore species by wet sieving over a cascade of nine sieves with mesh sizes ranging from 0.063 to 16 mm. MOF, dMEAN and dMED can be calculated directly from the results of sieve analysis, but cMEAN and cMED require a curve-fitting procedure. Across the whole sample size, dMEAN and cMEAN showed the highest correlation. The correlation between the respective MEAN and MED was higher for d than for c. As expected, MOF deviated most from the other measurements. Simulating different sieve sets resulted in a poor correlation between the results from the different sets in MOF and cMED, but a good correlation in dMEAN and cMEAN, suggesting that these latter measures can also be compared between studies that do not use identical sieve sets. As the calculation of dMEAN is comparatively simpler and less time-consuming than that of cMEAN, we propose the dMEAN as a standard for the description of a mean particle size value obtained from sieve analysis. For practical application, the good correlation of different simulated sieve sets indicates that sets with fewer sieves could be used in large-scale studies to reduce analytical workload.

Prolonged storage of epididymal spermatozoa does not affect their capacity to fertilise in vitro-matured domestic cat (Felis catus) oocytes when using ICSI.

The impact of different storage conditions of epididymal spermatozoa (including prolonged storage, cryopreservation and freeze-drying) on their fertilisation capacity was tested using intracytoplasmic sperm injection (ICSI). This kind of information is urgently needed when applying assisted reproductive technology to endangered felids in zoos. In particular, the utilisation of epididymal spermatozoa of castrated or deceased felids often requires time-consuming transportation and is therefore susceptible to loss of gamete quality. Sperm cells were stored at 4 °C for up to 72 h followed by cryopreservation or freeze-drying. Thawed motile and immotile spermatozoa were used for ICSI and the embryo cleavage rate was assessed 36 h after injection. A significant impact on the fertilisation rate of oocytes could only be detected when using immotile thawed or rehydrated spermatozoa. Cryopreservation or storage at 4 °C showed no influence. The simulation of transport conditions using domestic cat spermatozoa revealed that in vitro production of felid embryos with gametes from euthanised individuals is possible if testes are stored cool and arrive at the laboratory within 72 h. An essential prerequisite is the application of ICSI to achieve fertilisation even with single motile spermatozoa. Additional cryopreservation of spermatozoa after transportation is possible and will allow the establishment of a sperm bank for felids.

Phylogenetic constraints on digesta separation: Variation in fluid throughput in the digestive tract in mammalian herbivores.

The relevance of the mean retention time (MRT) of particles through the gastrointestinal tract (GIT) is well understood and MRT(particle)GIT is an important parameter in digestion models. Solute markers have been used to estimate MRT(solute)GIT (or 'fluid passage') in animals, but the relevance of this measure is less evident and is usually sought in its relation to MRT(particle)GIT. The ratio between the two measures indicates the degree of 'digesta washing', with little washing occurring at ratios of 1, aborad washing at ratios >1 (where the solute marker travels faster than the particle marker), and orad (retrograde) washing at ratios <1 (where the solute marker travels slower than the particle marker). We analysed digesta washing in a dataset of 98 mammalian species including man of different digestion types (caecum, colon and nonruminant foregut fermenters, and ruminants), controlling for phylogeny; a subset of 72 species allowed testing for the influence of food intake level. The results indicate that MRT(solute)GIT and the degree of digesta washing are related to digestion type, whereas variation in MRT(particle)GIT is influenced mainly by effects of body mass and food intake. Thus, fluid throughput and digesta washing emerge as important correlates of digestive anatomy. Most importantly, primates appear constrained to little digesta washing compared to non-primate mammalian herbivores, regardless of their digestion type. These results may help explain the absence of primates from certain herbivore niches and represent a drastic example of a physiologic limitation in a phylogenetic group. More experimental research is required to illuminate relative benefits and costs of digesta washing.

Mating system, feeding type and ex situ conservation effort determine life expectancy in captive ruminants.

Zoo animal husbandry aims at constantly improving husbandry, reproductive success and ultimately animal welfare. Nevertheless, analyses to determine factors influencing husbandry of different species are rare. The relative life expectancy (rLE; life expectancy (LE) as proportion of longevity) describes husbandry success of captive populations. Correlating rLE with biological characteristics of different species, reasons for variation in rLE can be detected. We analysed data of 166 901 animals representing 78 ruminant species kept in 850 facilities. The rLE of females correlated with the percentage of grass in a species' natural diet, suggesting that needs of species adapted to grass can be more easily accommodated than the needs of those adapted to browse. Males of monogamous species demonstrate higher rLE than polygamous males, which matches observed differences of sexual bias in LE in free-living populations and thus supports the ecological theory that the mating system influences LE. The third interesting finding was that rLE was higher in species managed by international studbooks when compared with species not managed in this way. Our method facilitates the identification of biological characteristics of species that are relevant for their husbandry success, and they also support ecological theory. Translating these findings into feeding recommendations, our approach can help to improve animal husbandry.

A longitudinal study on avian polyomavirus-specific antibodies in captive Spix's macaws (Cyanopsitta spixii).

Avian polyomavirus (APV) causes a range of disease syndromes in psittacine birds, from acute fatal disease to subclinical infections, depending on age, species, and other unidentified risk factors. To determine the prevalence of APV-specific antibodies in a captive population of Spix's macaws (Cyanopsitta spixii) in Quatar, 54 birds were tested by blocking enzyme-linked immunosorbent assay. A prevalence of 48.1% for APV antibodies, which indicates viral exposure, was found. Of 36 Spix's macaws that were serially tested over a period of 4 years, 50.0% were consistently positive, 36.1% were consistently negative, 5.5% had permanently declining antibody levels, and 2.8% showed variable results. By using polymerase chain reaction testing on whole blood samples, an apparent viremia was detected in 1 of 44 birds (2.3%), although contamination provides a likely explanation for this isolated positive result in a hand-reared chick. The white blood cell count was significantly higher in antibody-positive birds compared with antibody-negative birds (P < .05). Because antibody-positive and antibody-negative birds were housed together without a change in their respective antibody status, transmission of APV within the adult breeding population appeared to be a rare event.

Comparative ingestive mastication in domestic horses and cattle: a pilot investigation.

It is often assumed that horses chew food more intensively during ingestion than cattle, which - as ruminants - complete part of the mastication during rumination. This has been proposed as a reason for more robust mandibles, larger masseter insertion areas and larger masseter muscles in horses as compared to cattle and other grazing ruminants. In this study, we evaluate results of comparative feeding trials with three horses (338-629 kg) and three cows (404-786 kg), on four different roughages. Ingestion time (s/g dry matter) and chewing intensity (chews/g dry matter) differed among animals within a species, indicating an influence of body mass, and differed significantly between different forages. However, although numerical differences clearly suggest that horses have longer ingestion times and higher chewing intensities on high-fibre roughage than do cattle, this could not be proven in this dataset, most likely because of the small number of individuals sampled. Further studies are required to corroborate the suspected ingestive behaviour difference between equids and ruminants.

To chew or not to chew: fecal particle size in herbivorous reptiles and mammals.

A major difference between reptile and mammalian herbivores is that the former do not masticate their food. Actually, food particle size reduction by chewing is usually considered one of the adaptations facilitating the higher metabolic rates of mammals. However, quantitative comparisons of ingesta particle size between the clades have, to our knowledge, not been performed so far. We measured mean fecal particle size (MPS) in 79 captive individuals of 14 reptile herbivore species (tortoises, lizards, and Corucia zebrata) by wet sieving and compared the results with a mammalian dataset. MPS increased with body mass in both clades, but at a significantly higher level in reptiles. Limited evidence in free-ranging and captive individuals of Testudo hermanni indicates that in reptiles, the ability to crop food and food particle size significantly influence fecal particle size. The opportunistic observation of a drastic particle size difference between stomach and intestinal contents corroborates findings that in reptiles, in contrast to terrestrial mammals, significant ingesta particle size reduction does occur in the gastrointestinal tract, most likely owing to microbial action during very long ingesta retention. Whether behavioral adaptations to controlling ingesta particle size, such as deliberate small bite sizes, are adaptive strategies in reptiles remains to be investigated.

Comparative investigations on digestion in grazing (Ceratotherium simum) and browsing (Diceros bicornis) rhinoceroses.

Rhinoceroses represent the largest extant herbivores with extensive dietary specialization for plant groups like browse (black rhino Diceros bicornis) or grass (white rhino Ceratotherium simum). However, it is not clear to what extent such diet selection patterns are reflected in adaptations of digestive physiology of the respective feeding types. In this study, feeding trials with four black and five white rhinos were conducted in four zoos. The animals had ad libitum access to the same batch of grass hay (second cut; neutral detergent fiber (NDF) 63% dry matter (DM), crude protein 10.2% DM). Total intake, fecal N content, in vitro digestibility of NDF residues of feces, fecal particle size and mean retention time (MRT) of particles (Cr-mordanted fiber; 1-2mm) and fluid (Co-EDTA) were quantified. The average daily DM intake was 70+/-12 g/kg BW(0.75) for white and 73+/-10 g/kg BW(0.75) for black rhinos. In the in vitro fermentation test fecal NDF residues of black rhinos resulted in higher gas productions at fermentation times of 12 to 24h, indicating that white rhinos have a superior capacity to digest NDF. Average MRT for fluids and particles was 28+/-4h and 43+/-5h in white and 34+/-4h and 39+/-4h in black rhinos. The selectivity factor (SF=MRT(particle)/MRT(fluid)) was higher for white (1.5+/-0.2) than for black rhinos (1.2+/-0.1) (p=0.016). In a comparison of 12 ruminant and 3 rhino species, SF was correlated to percentage of grass in diet (R=0.75). Mean fecal particle size was higher in white (9.1+/-1.94 mm) than in black rhinos (6.1+/-0.79 mm) (p=0.016). The results demonstrate differences between white and black rhinos in terms of retention times and fiber digestibility. The more selective retention of particles by the white rhino corresponds with the higher digestion of fiber measured indirectly. Furthermore there is indication for a general pattern of high SF in grazing ruminants and rhinos. The difference in fecal particle size between both rhino species might be due to the considerable difference in body weight.

Relevance of management and feeding regimens on life expectancy in captive deer.

OBJECTIVE: To establish a demographic approach to facilitate the comparison of husbandry success for deer species in zoos and to test for factors that influence the performance of deer species in captivity. SAMPLE POPULATION: Data collected from 45,736 zoo-kept deer that comprised 31 species. PROCEDURES: Data had been collected by the International Species Information System during the last 3 decades on zoo-kept deer around the world. The relative life expectancy (rLE) of a species (ie, mean life expectancy as a proportion of the maximum recorded life span for that species) was used to describe zoo populations. The rLE (values between 0 and 1) was used to reflect the husbandry success of a species. RESULTS: A significant positive correlation was found between the rLE of a species and the percentage of grass in the natural diet of the species, suggesting that there are more problems in the husbandry of browsing than of grazing species. The 4 species for which a studbook (ie, record of the lineage of wild animals bred in captivity) was maintained had a high rLE, potentially indicating the positive effect of intensive breeding management. CONCLUSION AND CLINICAL RELEVANCE: The rLE facilitated the comparison of husbandry success for various species and may offer the possibility of correlating this quotient with other biological variables. Ultimately, identifying reasons for a low husbandry success in certain species may form the basis for further improvements of animal welfare in captivity.

Passage marker excretion in red kangaroo (Macropus rufus), collared peccary (Pecari tajacu) and colobine monkeys (Colobus angolensis, C. polykomos, Trachypithecus johnii).

Ruminants are characterized by an efficient particle-sorting mechanism in the forestomach (FRST) followed by selective rechewing of large food particles. For the nonruminating foregut fermenter pygmy hippo it was demonstrated that large particles are excreted as fast as, or faster than, the small particles. The same has been suggested for other nonruminating foregut fermenters. We determined the mean retention time of fluids and different-sized particles in six red kangaroos (Macropus rufus), seven collared peccaries (Pecari tajacu) and three colobine monkeys (Colobus angolensis, C. polykomos, Trachypithecus johnii). We fed Co-EDTA as fluid and mordanted fiber as particle markers (Cr, Ce). Mean (+ or - SD) total tract retention time for fluids, small and large particles was 14 + or - 2, 29 + or - 10 and 30 + or - 9 hr in red kangaroos, 26 + or - 2, 34 + or - 5 and 32 + or - 3 hr in collared peccaries and 57 + or - 17, 55 + or - 19 and 54 + or - 19 hr in colobine monkeys, respectively. Large and small particles were excreted simultaneously in all species. There was no difference in the excretion of fluids and particles in the colobine monkeys, in contrast to the other foregut fermenters. In the nonprimate, nonruminant foregut fermenters, the difference in the excretion of fluids and small particles decreases with increasing food intake. On the contrary, ruminants keep this differential excretion constant at different intake levels. This may be a prerequisite for the sorting of particles in their FRST and enable them to achieve higher food intake rates. The functional significance of differential excretion of fluids and particles from the FRST requires further investigations.

Physical characteristics of reticuloruminal contents of oxen in relation to forage type and time after feeding.

The stratification of reticulorumen (RR) contents has been described in domestic ruminants, but suspected to be absent in certain wild ruminant species. To investigate how such stratification could be quantified, we tested variables indicating stratification in three oxen with rumen cannulae, fed once daily for 3 h, one of three different forages (fresh grass FG, grass hay GH, lucerne hay LH). Ingesta samples from dorsal rumen (DR), ventral rumen (VR) and reticulum (RET) were taken at 0, 3, 6, 12 and 24 h after the last meal and analyzed for dry matter (DM) concentration, mean particle size and proportion of floating/sedimenting particles. Viscosity was measured in centrifuged RR fluid. There was no relevant variation in rumen fluid viscosity over time or with feeding regime. Dry matter content in DR was always higher than in VR or RET and increased after feeding. When LH and FG were fed, DR always contained larger particles than VR, except at 6 and 12 h of feeding GH. The proportion of floating particles was higher in DR than VR except at GH 6 h and GR between 6 and 12 h. The floating particle fraction mostly contained larger particles, except for GH 3 h in both DR and VR and for FG 3-12 h in DR. Changes in the distribution of particle size and proportion of floating particles over time comply with the theoretical course of fermentative digestion for different forages, but make these variables unreliable indicators of stratification in less controlled settings, such as investigations in wild ruminants. In contrast, viscosity of rumen fluids appears constant over time and feeding regime and might be used as a species-specific variable. The difference in DM content in DR and VR is a reliable indicator for RR contents stratification characterized by an oversaturation of the contents with fluid, leading to lower VR DM contents.

Changes in body weight, hematology and serum chemistry in captive plains viscachas (Lagostomus maximus) with presumptive diabetes type II after a diet change.

We report body weights (BW) and blood and serum analyses for 6 fully mature and 8 not-yet-mature captive plains viscachas before and 3, 6 and 9 months after switching from a low-fiber, high-energy diet to a high-fiber, low-energy diet. Initially, body weights, serum glucose, fructosamine and cholesterol levels were above the reference range in the fully mature animals. Furthermore, 4 of these animals had bilateral cataracts. After the diet change, these parameters dropped into the reference range. However, 9 months later, a slightly increased BW became evident again. The findings are consistent with a type II diabetes mellitus and underline the importance of dietary prevention.

The intraruminal papillation gradient in wild ruminants of different feeding types: Implications for rumen physiology.

Browsing and grazing ruminants are thought to differ in the degree their rumen contents are stratified-which may be due to different characteristics of their respective forages, to particular adaptations of the animals, or both. However, this stratification is difficult to measure in live animals. The papillation of the rumen has been suggested as an anatomical proxy for stratification-with even papillation indicating homogenous contents, and uneven papillation (with few and small dorsal and ventral papillae, and prominent papillae in the atrium ruminis) stratified contents. Using the surface enlargement factor (SEF, indicating how basal mucosa surface is increased by papillae) of over 55 ruminant species, we demonstrate that differences between the SEF(dorsal) or SEF(ventral) and the SEF(atrium) are significantly related to the percentage of grass in the natural diet. The more a species is adapted to grass, the more distinct this difference, with extreme grazers having unpapillated dorsal and ventral mucosa. The relative SEF(dorsal) as anatomical proxy for stratification, and the difference in particle and fluid retention in the rumen as physiological proxy for stratification, are highly correlated in species (n = 9) for which both kind of data are available. The results support the concept that the stratification of rumen contents varies among ruminants, with more homogenous contents in the more browsing and more stratified contents in the more grazing species.

More efficient mastication allows increasing intake without compromising digestibility or necessitating a larger gut: comparative feeding trials in banteng (Bos javanicus) and pygmy hippopotamus (Hexaprotodon liberiensis).

The digestion of plant material in mammalian herbivores basically depends on the chemical and structural composition of the diet, the mean particle size to which the forage is processed, and the ingesta retention time. These different factors can be influenced by the animal, and they can presumably compensate for each other. The pygmy hippopotamus, a non-ruminating foregut fermenter, has longer mean retention times than ruminants; however hippos do not achieve higher (fibre) digestibilities on comparable diets, which could be due to ineffective mastication. We performed feeding trials with six pygmy hippos (Hexaprotodon liberiensis) and six banteng cattle (Bos javanicus) on a grass diet. As predicted, both species achieved similar dry matter, organic matter, crude protein and gross energy digestibilities. However, neutral and acid detergent fibre digestibility was lower in pygmy hippos. Apparently, in these species, fibre digestibility was more influenced by particle size, which was larger in pygmy hippos compared to banteng, than by retention time. In spite of their higher relative food intake, the banteng in this study did not have greater relative gut fills than the hippos. Ruminants traditionally appear intake-limited when compared to equids, because feed particles above a certain size cannot leave the rumen. But when compared to nonruminating foregut fermenters, rumination seems to free foregut fermenters from an intrinsic food intake limitation. The higher energy intakes and metabolic rates in wild cattle compared to hippos could have life-history consequences, such as a higher relative reproductive rate.

No easy solution for the fractionation of faecal nitrogen in captive wild herbivores: results of a pilot study.

Faecal nitrogen (FN) concentration is used as a marker for habitat quality and digestive efficiency in free-ranging herbivores. In herbivores, FN can be separated into undigested plant N (analysed as the N concentration of the neutral detergent residue) and metabolic faecal N (MFN). It has been suggested that by differential analysis of the faecal fibre-bound N, the MFN fraction can be further split into a bacterial N and an endogenous N fraction [Hesta et al., Br. J. Nutr. 90 (2003) 1007]. We applied these methods to 96 faecal samples of 48 mammalian herbivore species from zoos. Species were grouped into coprophageous and non-coprophageous hindgut fermenters and ruminating and non-ruminating foregut fermenters. Diet was not controlled. The FN decreased with body mass, possibly reflecting higher proportions of concentrates in diets of smaller animals. The proportion of MFN increased with FN, indicating that higher quality food might enhance the gastrointestinal bacterial flora. The only outlier to this pattern was the lesser panda (Ailurus fulgens), confirming the low relevance of fermentative digestion in this herbivorous 'carnivore'. No relevant differences between the four digestion types were noted. The proportion of endogenous faecal N (32-80% of FN) was always higher than that of bacterial faecal N (7-30%), which contradicts basal understanding of herbivore digestive physiology. Thus, the method of Hesta et al. (2003) does not appear applicable to herbivores. While the results do not exclude the possibility that detailed differences might occur between digestion types, they indicate a high degree of similarity between herbivores that rely on bacterial fermentation, regardless of their digestion type, with respect to metabolic faecal losses.

Split-sample comparison of directional and liquid nitrogen vapour freezing method on post-thaw semen quality in white rhinoceroses (Ceratotherium simum simum and Ceratotherium simum cottoni).

To increase the quality of cryopreserved sperm in white rhinoceros, the liquid nitrogen vapour (LN vapour) freezing and the multi-thermal gradient directional freezing methods were compared. Sixteen white rhinoceros (Ceratotherium simum sp.) were electro-ejaculated. Semen samples were diluted with cryoextender (Tris, lactose, egg-yolk, DMSO) and aliquoted into straws for LN vapour freezing, and glass hollow tubes for directional freezing. The sperm quality was evaluated before and after freezing by assessing the following parameters: motility, morphologic state, acrosomal integrity and plasma membrane function and integrity (i.e. sperm viability) as defined by the hypo-osmotic swelling. Directional freezing improved the sperm viability by 5.6% (p<0.005), progressive motility score by 34.7% and sperm motility index (SMI) by 8.1% (p<0.005) versus LN vapour freezing. When data was categorized into groups of low (<19%), moderate (20-39%) and high (>40%) percentages of morphologically normal, directional freezing (DF) resulted in 31.4% less abnormal acrosomes for the low quality group as well as 18.7% increase in intact acrosomes and 10.9% increase in motility for the high quality group compared to LN vapour freezing (LN) (p<0.01, p<0.03, p<0.01, respectively). LN showed a significant reduction in sperm head volume (5.7%, p<0.05) compared to the prefreeze; whereas, no significant reduction in head volume was demonstrated after DF. Several additives (xanthenuric acid, cytochalasin D, potassium, EDTA) to the basic cryoextender provided no significant improvement in spermatozoal survival after directional freezing. In conclusion, directional freezing proved to facilitate higher gamete survival compared to LN vapour freezing. This is especially effective in ejaculates of low sperm quality and is important in endangered species where high quality semen donors are often not accessible. These results suggest that directional freezing could be valuable particularly for species with limited freezability of spermatozoa.

Physical characteristics of rumen contents in two small ruminants of different feeding type, the mouflon (Ovis ammon musimon) and the roe deer (Capreolus capreolus).

In domestic ruminants, the stratification of forestomach contents - the results of flotation and sedimentation processes - is an important prerequisite for the selective particle retention in this organ. A series of anatomical and physiological measurements suggests that the degree of this stratification varies between browsing and grazing wild ruminants. We investigated the forestomach contents of free-ranging mouflon and roe deer shot during regular hunting procedures. There was no difference between the species in the degree by which forestomach ingesta separated according to size due to buoyancy characteristics in vitro. However, forestomach fluid of roe deer was more viscous than that of mouflon, and no difference in moisture content was evident between the dorsal and the ventral rumen in roe deer, in contrast to mouflon. Hence, the forestomach milieu in roe deer appears less favourable for gas or particle separation due to buoyancy characteristics. These findings are in accord with notable differences in forestomach papillation between the two species. In roe deer, particle separation is most likely restricted to the reticulum, whereas in mouflon, the whole rumen may pre-sort particles to a higher degree. The results suggest that differences in forestomach physiology may occur across ruminant species.

Higher masseter muscle mass in grazing than in browsing ruminants.

Using cranioskeletal measurements, several studies have generated evidence that grazing ruminants have a more pronounced mastication apparatus, in terms of muscle insertion areas and protuberances, than browsing ruminants, with the resulting hypothesis that grazers should have larger, heavier chewing muscles than browsers. However, the only investigation of this so far [Axmacher and Hofmann (J Zool 215:463-473, 1988)] did not find differences between ruminant feeding types in the masseter muscle mass of 22 species. Here, we expand the dataset to 48 ruminant species. Regardless of phylogenetic control in the statistical treatment, there was a significant positive correlation of body mass and masseter mass, and also a significant association between percent grass in the natural diet and masseter mass. The results support the concept that ruminant species that ingest more grass have relatively larger masseter muscles, possibly indicating an increased requirement to overcome the resistance of grass forage. The comparative chewing resistance of different forage classes may represent a rewarding field of ecophysiological research.