Investigations on the stratification of forestomach contents in ruminants: an ultrasonographic approach

The degree of stratification in the reticulorumen contents is considered a major explanatory factor for other morphological and physiological differences between ruminant feeding types. However, information on reticulorumen (RR) contents is usually gathered from dead animals or indirect such as passage kinetics. We hypothesized that, although the contents of the gastrointestinal tract can usually not be evaluated by ultrasonography, the three typical layers of stratified RR contents (gas dome, fibre mat, fluid) can be demonstrated by this technique. In three domestic cows, the gas dome in the cows’ rumens could be demonstrated sonographically by reverberation lines running in parallel to the line demarcating the rumen mucosa, as is typical for the sonographic image of large, gas-filled spaces. More ventrally, the area behind the rumen mucosa showed typical pattern indicating gaseous inclusions, corresponding to the fibre mat inside the rumen. Further ventrally, in two of the animals, the area behind the demarcation line appeared dark without reverberation lines, as is typical for large fluid-filled spaces. When the technique was applied to a captive, habituated, browse-fed moose, no gas dome could be demonstrated, supporting the hypothesis that the reticulorumen contents of browsers are less stratified. The results of this study indicate that sonography represents a useful tool for the demonstration of RR contents stratification in live animals.     

Methane mitigation in ruminants: from microbe to the farm scale

Decreasing enteric methane (CH4) emissions from ruminants without altering animal production is desirable both as a strategy to reduce global greenhouse gas (GHG) emissions and as a means of improving feed conversion efficiency. The aim of this paper is to provide an update on a selection of proved and potential strategies to mitigate enteric CH4 production by ruminants. Various biotechnologies are currently being explored with mixed results. Approaches to control methanogens through vaccination or the use of bacteriocins highlight the difficulty to modulate the rumen microbial ecosystem durably. The use of probiotics, i.e. acetogens and live yeasts, remains a potentially interesting approach, but results have been either unsatisfactory, not conclusive, or have yet to be confirmed in vivo. Elimination of the rumen protozoa to mitigate methanogenesis is promising, but this option should be carefully evaluated in terms of livestock performances. In addition, on-farm defaunation techniques are not available up to now. Several feed additives such as ionophores, organic acids and plant extracts have also been assayed. The potential use of plant extracts to reduce CH4 is receiving a renewed interest as they are seen as a natural alternative to chemical additives and are well perceived by consumers. The response to tannin- and saponin-containing plant extracts is highly variable and more research is needed to assess the effectiveness and eventual presence of undesirable residues in animal products. Nutritional strategies to mitigate CH4 emissions from ruminants are, without doubt, the most developed and ready to be applied in the field. Approaches presented in this paper involve interventions on the nature and amount of energy-based concentrates and forages, which constitute the main component of diets as well as the use of lipid supplements. The possible selection of animals based on low CH4 production and more likely on their high efficiency of digestive processes is also addressed. Whatever the approach proposed, however, before practical solutions are applied in the field, the sustainability of CH4 suppressing strategies is an important issue that has to be considered. The evaluation of different strategies, in terms of total GHG emissions for a given production system, is discussed.     

Effect of the corn silage to grass silage ratio and feed particle size of diets for ruminants on the ruminal Bacteroides-Prevotella community in vitro

This study examined whether different corn silage to grass silage ratios in ruminant rations and different grinding levels of the feed affect the composition of the ruminal Bacteroides-Prevotella community in vitro. Three diets, composed of 10% soybean meal as well as of different corn silage and grass silage proportions, were ground through 1 mm or 4 mm screened sieves and incubated in a semi-continuous rumen simulation system. On day 14 of the incubation microbes were harvested by centrifugation from the liquid effluent of fermenter vessels. Microbial DNA was extracted for single strand conformation polymorphism (SSCP) analysis of 16S rRNA genes followed by sequencing of single SSCP bands. Fluorescence in situ hybridization (FISH) and real-time quantitative (q) PCR were used to quantify differences in the relative abundance of Bacteroides-Prevotella and Prevotella bryantii. SSCP profiles revealed a significant influence of the forage source as well as of the feed particle size on the community structure of the Bacteroides-Prevotella group. Different, phylogenetically distinct, so far uncultured Prevotella species were detected by sequence analysis of several treatment-dependent occurring SSCP bands indicating different nutritional requirements of these organisms for growth. No quantitative differences in the occurrence of Bacteroides-Prevotella-related species were detected between diets by FISH with probe BAC303. However, real-time qPCR data revealed a higher abundance of P. bryantii with increasing grass silage to corn silage ratio, thus again indicating changes within the community composition of the Bacteroides-Prevotella group. As P. bryantii possesses high proteolytic activity its higher abundance may have been caused by the higher contents of crude protein in the grass silage containing diets. To conclude, results of this study show an influence of the forage source on the ruminal community of Bacteroides-Prevotella. Furthermore, they suggest an effect of the feed particle size on this bacterial group.     

Phylogenetic comparative methods: Harnessing the power of species diversity to investigate welfare issues in captive wild animals

This paper reviews a way of investigating health and welfare problems in captive wild animals (e.g., those in zoos, aviaries, aquaria, or aquaculture systems) that has great potential, but to date has been little used: systematically comparing species with few or no health and welfare issues to those more prone to problems. Doing so empirically pinpoints species‐typical welfare risk and protective factors (such as aspects of their natural behavioral biology): information which can then be used to help prevent or remedy problems by suggesting new ways to improve housing and husbandry, and by identifying species intrinsically best suited to captivity. We provide a detailed, step‐by‐step “how to” guide for researchers interested in using these techniques, including guidance on how to statistically control for the inherent similarities shared by related species: an important concern because simple, cross‐species comparisons that do not do this may well fail to meet statistical assumptions of non‐independence. The few relevant studies that have investigated captive wild animals’ welfare problems using this method are described. Overall, such approaches reap value from the great number and diversity of species held in captivity (e.g., the many thousands of species held in zoos); can yield new insights from existing data and published results; render previously intractable welfare questions (such as “do birds need to fly?” or “do Carnivora need to hunt?”) amenable to study; and generate evidence‐based principles for integrating animal welfare into collection planning.     

Changes in pig diet particle size profile and nutrient content during on-farm storage and distribution to the feeders

The present study assessed the effect of silo emptying and feed transport by conveyor systems on particle size and nutrient content of the feed delivered to the pigs. Experiment 1 sampled feed from four feeders along the conveyor system of two barns. Samples were taken immediately after filling the feed silo (Begin) and when the silo was almost empty (End). In Experiment 2, three barns with drag-type conveyors, three with auger-type conveyors and two with spiral-type conveyors were sampled. Along the different conveyors, samples at 10, 20, 50 and 85 m distance from the feed silo were taken from the feeders. In each barn, sampling was repeated for two subsequent batches of feed delivered. In all samples, particle size profile was determined and nutrient content was analysed. In Experiment 2, mineral content was also determined. In Experiment 1, the size of the different particle fractions decreased from Begin to End. An interaction (p < 0.05) between sampling time and conveyor type was detected for the 10% smallest particles. In Experiment 2, an effect of sampling time on the 10% largest particles was detected (p < 0.05). The effect of sampling time on several nutrients was observed in Experiments 1 and 2, but the affected nutrients differed between both experiments. Results implied that it was mainly emptying of the silo that affected mash particle size profile and nutrient content. The potential impact of these changes on pig performance requires further investigation.     

Influence of the maize silage to grass silage ratio and feed particle size of rations for ruminants on the community structure of ruminal Firmicutes in vitro

Aims: To investigate the effect of the forage source and feed particle size (FPS) in ruminant rations on the composition of the ruminal Firmicutes community in vitro. Methods and Results: Three diets, varying in maize silage to grass silage ratio and FPS, were incubated in a rumen simulation system. Microbial samples were taken from the liquid fermenter effluents. Microbial community analysis was performed by 16S rRNA‐based techniques. Clostridia‐specific single‐strand conformation polymorphism profiles revealed changes of the community structure in dependence on both factors tested. The coarse grass silage–containing diets seemed to enhance the occurrence of different Roseburia species. As detected by real‐time quantitative PCR, Ruminococcus albus showed a higher abundance with decreasing FPS. A slightly lower proportion of Bacilli was found with increasing grass silage to maize silage ratio by fluorescence in situ hybridization (FISH). In contrast, a slightly higher proportion of bacterial species belonging to the Clostridium‐clusters XIV a and b was detected by FISH with increasing grass silage contents in the diet. Conclusions: The ruminal Firmicutes community is affected by the choice of the forage source and FPS. Significance and Impact of the Study: This study supplies fundamental knowledge about the response of ruminal microbial communities to changing diets. Moreover, the data suggest a standardization of grinding of feeds for in vitro studies to facilitate the comparison of results of different laboratories.     

The generation of diesel exhaust particle aerosols from a bulk source in an aerodynamic size range similar to atmospheric particles

The influence of diesel exhaust particles (DEP) on the lungs and heart is currently a topic of great interest in inhalation toxicology. Epidemiological data and animal studies have implicated airborne particulate matter and DEP in increased morbidity and mortality due to a number of cardiopulmonary diseases including asthma, chronic obstructive pulmonary disorder, and lung cancer. The pathogeneses of these diseases are being studied using animal models and cell culture techniques. Real-time exposures to freshly combusted diesel fuel are complex and require significant infrastructure including engine operations, dilution air, and monitoring and control of gases. A method of generating DEP aerosols from a bulk source in an aerodynamic size range similar to atmospheric DEP would be a desirable and useful alternative. Metered dose inhaler technology was adopted to generate aerosols from suspensions of DEP in the propellant hydrofluoroalkane 134a. Inertial impaction data indicated that the particle size distributions of the generated aerosols were trimodal, with count median aerodynamic diameters less than 100 nm. Scanning electron microscopy of deposited particles showed tightly aggregated particles, as would be expected from an evaporative process. Chemical analysis indicated that there were no major changes in the mass proportion of 2 specific aromatic hydrocarbons (benzo[a]pyrene and benzo[k]fluoranthene) in the particles resulting from the aerosolization process.     

The importance of scale in comparative microbiome research: New insights from the gut and glands of captive and wild lemurs

Research on animal microbiomes is increasingly aimed at determining the evolutionary and ecological factors that govern host–microbiome dynamics, which are invariably intertwined and potentially synergistic. We present three empirical studies related to this topic, each of which relies on the diversity of Malagasy lemurs (representing a total of 19 species) and the comparative approach applied across scales of analysis. In Study 1, we compare gut microbial membership across 14 species in the wild to test the relative importance of host phylogeny and feeding strategy in mediating microbiome structure. Whereas host phylogeny strongly predicted community composition, the same feeding strategies shared by distant relatives did not produce convergent microbial consortia, but rather shaped microbiomes in host lineage‐specific ways, particularly in folivores. In Study 2, we compare 14 species of wild and captive folivores, frugivores, and omnivores, to highlight the importance of captive populations for advancing gut microbiome research. We show that the perturbational effect of captivity is mediated by host feeding strategy and can be mitigated, in part, by modified animal management. In Study 3, we examine various scent‐gland microbiomes across three species in the wild or captivity and show them to vary by host species, sex, body site, and a proxy of social status. These rare data provide support for the bacterial fermentation hypothesis in olfactory signal production and implicate steroid hormones as mediators of microbial community structure. We conclude by discussing the role of scale in comparative microbial studies, the links between feeding strategy and host–microbiome coadaptation, the underappreciated benefits of captive populations for advancing conservation research, and the need to consider the entirety of an animal's microbiota. Ultimately, these studies will help move the field from exploratory to hypothesis‐driven research.     

Understanding the melanocyte distribution in human epidermis: an agent-based computational model approach

The strikingly even color of human skin is maintained by the uniform distribution of melanocytes among keratinocytes in the basal layer of the human epidermis. In this work, we investigated three possible hypotheses on the mechanism by which the melanocytes and keratinocytes organize themselves to generate this pattern. We let the melanocyte migration be aided by (1) negative chemotaxis due to a substance produced by the melanocytes themselves, or (2) positive chemotaxis due to a substance produced by keratinocytes lacking direct physical contact with a melanocyte, or (3) positive chemotaxis due to a substance produced by keratinocytes in a distance-to-melanocytes dependent manner. The three hypotheses were implemented in an agent-based computational model of cellular interactions in the basal layer of the human epidermis. We found that they generate mutually exclusive predictions that can be tested by existing experimental protocols. This model forms a basis for further understanding of the communication between melanocytes and other skin cells in skin homeostasis.     

Characterizing biological aerosol in a chamber: an approach to estimation of viable organisms in a single biological particle

There are practical and valid reasons topresent biological field trial referenceresults as agent containing particles per literof air (ACPLA). However, workers in biologicalaerosol research have a need to know how manyviable individual organisms are contained in asingle particle of a given diameter. Anecdotalevidence may exist suggesting that the task hasbeen accomplished but without a way toreplicate the measurements, it is difficult toaccept unsubstantiated claims. It is verydifficult to declare a finite number thatsatisfies all the experimental requirements, asthe problem is a statistical probability issue. This paper describes a method for estimatingthe number with practical instructions forreplicating the work in other laboratories.The test aerosol was contained in a 90 m³chamber at concentrations as low as 5 ACPLA. Amodified version of slit sampler collectedviable particles. A statistical method was usedto demonstrate sampling predictability at 95%confidence level. By using glass fiber filtersmounted in a dichotomous sampler, samplingefficiencies were estimated for a variety ofcommon aerosol collectors. The accumulated datapermitted the estimation of the number ofviable spores per particle. For a 2.5 to4 µm particle, arguments have beenpresented for considering 4.5 as the mostprobable ACPLA value.     

Heritability estimates from genomewide relatedness matrices in wild populations: Application to a passerine,using a small sample size

Genomic developments have empowered the investigation of heritability in wild populations directly from genomewide relatedness matrices (GRM). Such GRM‐based approaches can in particular be used to improve or substitute approaches based on social pedigree (PED‐social). However, measuring heritability from GRM in the wild has not been widely applied yet, especially using small samples and in nonmodel species. Here, we estimated heritability for four quantitative traits (tarsus length, wing length, bill length and body mass), using PED‐social, a pedigree corrected by genetic data (PED‐corrected) and a GRM from a small sample (n = 494) of blue tits from natural populations in Corsica genotyped at nearly 50,000 filtered SNPs derived from RAD‐seq. We also measured genetic correlations among traits, and we performed chromosome partitioning. Heritability estimates were slightly higher when using GRM compared to PED‐social, and PED‐corrected yielded intermediate values, suggesting a minor underestimation of heritability in PED‐social due to incorrect pedigree links, including extra‐pair paternity, and to lower information content than the GRM. Genetic correlations among traits were similar between PED‐social and GRM but credible intervals were very large in both cases, suggesting a lack of power for this small data set. Although a positive linear relationship was found between the number of genes per chromosome and the chromosome heritability for tarsus length, chromosome partitioning similarly showed a lack of power for the three other traits. We discuss the usefulness and limitations of the quantitative genetic inferences based on genomic data in small samples from wild populations.     

Singlet oxygen-mediated signaling in plants: moving from flu to wild type reveals an increasing complexity

Singlet oxygen (¹O₂)-mediated signaling has been established in the conditional fluorescent (flu) mutant of Arabidopsis. In the dark, the flu mutant accumulates free protochlorophyllide (Pchlide), a photosensitizer that in the light generates ¹O₂. The release of ¹O₂ leads to growth inhibition of mature plants and bleaching of seedlings. These ¹O₂-mediated responses depend on two plastid proteins, EXECUTER (EX) 1 and 2. An ex1/ex2/flu mutant accumulates in the dark Pchlide and upon illumination generates similar amounts of ¹O₂ as flu, but ¹O₂-mediated responses are abrogated in the triple mutant. The ¹O₂- and EX-dependent signaling pathway operates also in wild type placed under light stress. However, it does not act alone as in flu, but interacts with other signaling pathways that modulate ¹O₂-mediated responses. Depending on how severe the light stress is, ¹O₂- and EX-dependent signaling may be superimposed by ¹O₂-mediated signaling that does not depend on EX and is associated with photo-oxidative damage. Because of its high reactivity and short half-life, ¹O₂ is unlikely to be a signal that is translocated across the chloroplast envelope, but is likely to interact with other plastid components close to its site of production and to generate more stable signaling molecules during this interaction. Depending on the site of ¹O₂ production and the severity of stress, different signaling molecules may be expected that give rise to different ¹O₂-mediated responses.     

A locus-wide approach to assessing variation in the avian MHC: the B-locus of the wild turkey

Studies of major histocompatibility complex (MHC) diversity in non-model vertebrates typically focus on structure and sequence variation in the antigen-presenting loci: the highly variable and polymorphic class I and class IIB genes. Although these studies provide estimates of the number of genes and alleles/locus, they often overlook variation in functionally related and co-inherited genes important in the immune response. This study utilizes the sequence of the MHC B-locus derived from a commercial turkey to investigate MHC variation in wild birds. Sequences were obtained for nine interspersed MHC amplicons (non-class I/II) from each of 40 birds representing 3 subspecies of wild turkey (Meleagris gallopavo). Analysis of aligned sequences identified 238 single-nucleotide variants approximately one-third of which had minor allele frequencies >0.2 in the sampled birds. PHASE analysis identified 70 prospective MHC haplotypes in the wild turkeys, whereas a combined analysis with commercial birds identified almost 100 haplotypes in the species. Denaturing gradient gel electrophoresis (DGGE) of the class IIB loci was used to test the efficacy of single-nucleotide polymorphism (SNP) haplotyping to capture locus-wide variation. Diversity in SNP haplotypes and haplotype sharing among individuals was directly reflected in the DGGE patterns. Utilization of a reference haplotype to sequence interspersed regions of the MHC has significant advantages over other methods of surveying diversity while identifying high-frequency SNPs for genotyping. SNP haplotyping provides a means to identify both divergent haplotypes and homozygous individuals for assessment of immunological variation in wild and domestic populations.     

Latitudinal variation in the shape of the species body size distribution: an analysis using freshwater fishes

Many taxonomic and ecological assemblages of species exhibit a right-skewed body size-frequency distribution when characterized at a regional scale. Although this distribution has been frequently described, factors influencing geographic variation in the distribution are not well understood, nor are mechanisms responsible for distribution shape. In this study, variation in the species body size-frequency distributions of 344 regional communities of North American freshwater fishes is examined in relation to latitude, species richness, and taxonomic composition. Although the distribution of all species of North American fishes is right-skewed, a negative correlation exists between latitude and regional community size distribution skewness, with size distributions becoming left-skewed at high latitudes. This relationship is not an artifact of the confounding relationship between latitude and species richness in North American fishes. The negative correlation between latitude and regional community size distribution skewness is partially due to the geographic distribution of families of fishes and apparently enhanced by a nonrandom geographic distribution of species within families. These results are discussed in the context of previous explanations of factors responsible for the generation of species size-frequency distributions related to the fractal nature of the environment, energetics, and evolutionary patterns of body size in North American fishes.     

Behavioral solutions to breeding cheetahs in captivity: Insights from the wild

Knowledge of cheetahs' behavior is increasingly seen as the key to solving the mystery of cheetahs' poor breeding performance in captivity. In the absence of zoos' maintaining systematic records of individuals' behavior during introductions, behavior of free-living animals can be informative. In the wild, most female cheetahs probably mate with males living in small groups or coalitions; thus, zoos may benefit from replicating these social conditions, provided injuries can be minimized. Relations between free-living coalition members are amicable, and escalated aggression was never witnessed during 4 years of observation. Some antagonism was seen in newly formed trios, although this had disappeared in longer established coalitions. Minor aggression occurred over carcasses, being greater at small and intermediate sized kills than at large ones. In the presence of females, mild intramale aggression was only seen within 1 of 7 coalitions. Free-living females showed similar rates of behavior in the presence of different numbers of males, aside from more frequent growling at large groups. These findings suggest that captive institutions should be less nervous about housing male cheetahs together and introducing females to groups of males for purposes of breeding. © 1993 Wiley-Liss, Inc.     

The ability of konjac-glucomannan to suppress spontaneously occurring dermatitis in NC/Nga mice depends upon the particle size

Konjac glucomannan (GM) is a well-known dietary fiber with various beneficial functions, which the higher viscous GM is proposed to have the stronger potencies. Application of highly viscous GM to various food industries is limited for the sake of long elapsed time required to solubilize the GM. Thus, the exploration into any physiological function of low-viscous GM remained undone for a long time. Male 4-week-old NC/Nga mice, a model of atopic dermatitis (AD), were fed for 8 weeks on diets containing 5% each of a high-viscous GM powder, ordinary konjac or highly purified GM, and a low-viscous GM powder, pulverized GM or re-granulated fine GM. Striking suppression to the aggravation of dermatitis and the increases in scratching behaviors and IgE levels was significantly observed only in mice fed on the pulverized GM diet, but not in mice fed on the other GM diets or a control diet. We concluded that the low-viscous GM possesses a novel immunoregulatory function and the intensity of the function depends upon the particle size rather than the viscosity.     

大兴安岭中段森林土壤的黑碳含量及其在不同粒级中的分布

土壤黑碳由于具有生物化学惰性被视为土壤稳定碳库的主要成分.本文量化了大兴安岭中段森林土壤黑碳,分析了黑碳在各粒级内的分布,并探讨了黑碳的稳定机制及其在土壤碳库中的重要性.结果表明: 土壤黑碳表聚现象明显,表层黑碳占总剖面(64 cm)的68.7%,随着土层加深黑碳含量显著降低,但黑碳占有机碳的比例却呈现上升趋势.气候条件影响大兴安岭地区土壤黑碳的分布,相对寒冷和湿润的气候条件提高了土壤固持黑碳的潜力;土壤黑碳在各粒级内所占比例表现为黏粒>粉粒>细砂>粗砂,尽管黏粒中黑碳含量最高,并随土层深度增加而升高,但黏粒中黑碳占有机碳的比例却无明显变化,黑碳/有机碳的升高主要源于细砂与粉粒中黑碳的增加;黑碳的生物化学惰性是表层黑碳的主要稳定机制,而深层的黑碳除了其自身抗性外,黏粒矿物的化学保护发挥着重要作用;黑碳不仅作为稳定碳库的主要成分,在颗粒有机碳组分中仍占相当大的比例,因此黑碳的存在提高了土壤稳定性碳储量与碳汇能力.     

Transition from a random to an ideal free to an ideal despotic distribution: the effect of individual difference in growth

Individual differences in growth can lead to a monopolistic form of food competition. We studied the long-term transition in the mode of competition and the distribution of individuals between food patches of the cloned salmonid fish, Oncorhynchus masou ishikawae, in the laboratory. This transition was accompanied by growth depensation, i.e., the increase over time in the variance of size between individuals resulting from the differences in individual growth rates. The 120-cm experimental tanks were divided into two compartments (patches) between which an opaque partition was placed. Fish were able to move freely between the patches and therefore were able to assess the patch quality using long-term memory, but they were not able to see the food input in the other patch directly. The distribution between the two food patches, the amount of food gained, and the growth and the agonistic behavior of four groups of six individuals were observed over 4 weeks. We found that (1) within-group variation in body weight increased with time; (2) on average, the better patch was used by more individuals than predicted by a random distribution but fewer individuals than predicted by an ideal free distribution, and (3) the distribution and pattern of resource use by the fish changed over the 4-week experimental period from a random distribution to an ideal free distribution and finally to an ideal despotic distribution. We suggest that growth depensation causes the long-term change in the spatial distribution and pattern of resource use by competitors. Received: December 19, 2000 / Accepted: March 19, 2001