Proteomics: A new tool in bovine claw disease research

Claw horn disruption (CHD) is a common underlying cause of lameness in dairy cattle which leads to compromised animal welfare and production losses. Despite an intense research effort over the last two decades, progress in reducing the prevalence of lameness due to CHD has been limited. In addition to current research strategies there is a need to develop novel approaches and methods that expand understanding of the disease mechanisms involved in CHD. The objectives of the present study were to explore the potential of liquid chromatography tandem mass spectrometry (LC-MS/MS) in mapping protein expression in three different bovine claw tissues, and to provide a relevant functional annotation of the proteins characterized in these tissues. LC-MS/MS was used to characterize protein expression in coronary band skin (C), claw dermal (D) and lamellar (L) tissues from two heifers. A total of 388 different proteins were identified, with 146 proteins available for identification in C, 279 proteins in D and 269 proteins in L. A functional annotation of the identified proteins was obtained using the on-line Blast2GO tool. Three hundred and sixteen of the identified proteins could be subsequently grouped manually to one or more of five major functional groups related to metabolism, cell structure, immunity, apoptosis and angiogenesis. These were chosen to represent basic cell functions and biological processes potentially involved in the pathogenesis of CHD. The LC-MS/MS-based proteomic analysis presented here is the largest published survey, so far, of the bovine claw tissue proteome.     

Intermittent Stress in Pigs: Behavioural and Pituitary-Adrenocortical Reactivity

Abstract

Ninetysix pigs, half females and half castrated males from 12 litters, were housed in 24 groups of 4 litter mates (2 females, 2 castrates). The pigs were weaned at 4 weeks of age and from the age of 115 days half of the groups were subjected to a schedule of unpredictable, inescapable electroshocks for 33 days, and half served as control. The adrenocortical reactivity to ACTH stimulation and to an emotional stressor (the procedure associated with initiation of blood collection) was measured in females, whereas the behavioural and pituitary-adrenocortical reactivity to an open field test was measured in castrates, as was the behavioural reactivity in a social test. Intermittent stress increased adrenocortical reactivity to ACTH stimulation significantly. This effect was present within the first week of intermittent stress (p < 0.06), but no difference was found after 4–5 weeks of stress. In contrast, the adrenocortical reactivity to additional emotional stressors was unaffected by stress after one week of intermittent stress, but increased after 4–5 weeks. In the open field test, centre location increased after one week of intermittent stress, whereas exploration decreased and walking and ambulation increased after 4–5 weeks of stress. In addition, 4–5 weeks of intermittent stress decreased aggressiveness in a social test. In conclusion, the change in reactivity to an additional acute stressor during intermittent stress appeared in different variables and contexts at different times during the course of intermittent stress. Thus, assessment of stress based on changed reactivity has to include several distinct measures of behavioural and hormonal reactivity.     

Impacts of prenatal nutrition on animal production and performance: a focus on growth and metabolic and endocrine function in sheep

The concept of foetal programming(FP) originated from human epidemiological studies, where foetal life nutrition was linked to health and disease status later in life. Since the proposal of this phenomenon, it has been evaluated in various animal models to gain further insights into the mechanisms underlying the foetal origins of health and disease in humans. In FP research, the sheep has been quite extensively used as a model for humans. In this paper we will review findings mainly from our Copenhagen sheep model, on the implications of late gestation malnutrition for growth, development, and metabolic and endocrine functions later in life, and discuss how these implications may depend on the diet fed to the animal in early postnatal life. Our results have indicated that negative implications of foetal malnutrition, both as a result of overnutrition and, particularly, late gestation undernutrition, can impair a wide range of endocrine functions regulating growth and presumably also reproductive traits. These implications are not readily observable early in postnatal life, but are increasingly manifested as the animal approaches adulthood. No intervention or cure is known that can reverse this programming in postnatal life. Our findings suggest that close to normal growth and slaughter results can be obtained at least until puberty in animals which have undergone adverse programming in foetal life, but manifestation of programming effects becomes increasingly evident in adult animals.Due to the risk of transfer of the adverse programming effects to future generations, it is therefore recommended that animals that are suspected to have undergone adverse FP are not used for reproduction. Unfortunately, no reliable biomarkers have as yet been identified that allow accurate identification of adversely programmed offspring at birth,except for very low or high birth weights, and, in pigs, characteristic changes in head shape(dolphin head). Future efforts should be therefore dedicated to identify reliable biomarkers and evaluate their effectiveness for alleviation/reversal of the adverse programming in postnatal life. Our sheep studies have shown that the adverse impacts of an extreme, high-fat diet in early postnatal life, but not prenatal undernutrition, can be largely reversed by dietary correction later in life. Thus, birth(at term) appears to be a critical set point for permanent programming in animals born precocial,such as sheep. Appropriate attention to the nutrition of the late pregnant dam should therefore be a priority in animal production systems.     

Using acoustic telemetry and snorkel surveys to study diel activity and seasonal migration of round goby (Neogobius melanostomus) in an estuary of the Western Baltic Sea

In just two decades, round goby Neogobius melanostomus (Pallas) has dispersed throughout most of the Baltic and the North American Great Lakes. It thrives in shallow sheltered areas, where it is impacting native fauna. In Denmark, researchers, governmental institutions and fishers have joined in an effort to establish a sustainable round goby fishery. To do so, basic information about the movement of round goby is essential, so the movements of 50 round gobies were tracked using acoustic telemetry. The results revealed that activity took place mainly at night and seasonal migrations were common. The most frequent migration behaviour observed was a seaward return migration during winter, which was further confirmed by a snorkel survey. Tracking data also revealed riverine upstream dispersal. The results demonstrate that the round goby can be mapped using telemetry, and it is suggested that fishing for round goby may improve efficiency by targeting seasonal migration corridors.     

The “soil microbial loop” is not always needed to explain protozoan stimulation of plants

Protozoa stimulate plant growth, but we do not completely understand the underlying mechanisms, and different hypotheses seek to explain this phenomenon. To test these hypotheses, we grew the grass Yorkshire fog (Holcus lanatus) in pots with soil, which contained either (1) no organisms but bacteria – or (2) bacteria and protozoa. Half of the pots received a glucose treatment so as to mimic an additional root exudation. We measured plant growth and plant nitrogen uptake, along with various microbial pools and processes that support plant growth. Protozoan presence significantly enhanced soil nitrogen mineralization, plant nitrogen uptake from organic nitrogen sources, plant nitrogen content, and plant growth. By contrast, we found no evidence that glucose addition, mimicking root exudation, increased soil nitrogen availability and plant nitrogen uptake. Moreover, although protozoan presence affected bacterial community structure, it did not affect the proportion of IAA-producing bacteria in the community or plant root morphology. These results refute the “soil microbial loop” hypotheses, which suggest that protozoan stimulation of plant growth results from complex interactions between plant roots, bacteria and protozoa. Our experiment thus favours the simple explanation that increased nitrogen availability is the key factor behind the positive protozoan effect on plant growth. To exploit natural resources in an efficient and environmentally friendly way, we need to understand in detail the functioning of ecosystems. This study stresses that to achieve this, it is still urgent, besides investigating intricate food-web and signal compound interactions, also to focus on the basic stoichiometric and energetic aspects of organisms.     

Static site indices from different national forest inventories: harmonization and prediction from site conditions

Key message

Static site indices determined from stands’ top height are derived from different forest inventory sources with height and age information and thus enable comparisons and modeling of a species’ productivity encompassing large environmental gradients.

Context

Estimating forest site productivity under changing climate requires models that cover a wide range of site conditions. To exploit different inventory sources, we need harmonized measures and procedures for the productive potential. Static site indices (SI) appear to be a good choice.

Aims

We propose a method to derive static site indices for different inventory designs and apply it to six tree species of the German and French National Forest Inventory (NFI). For Norway spruce and European beech, the climate dependency of SI is modeled in order to estimate trends in productivity due to climate change.

Methods

Height and age measures are determined from the top diameters of a species at a given site. The SI is determined for a reference age of 100 years.

Results

The top height proves as a stable height measure that can be derived harmoniously from German and French NFI. The boundaries of the age-height frame are well described by the Chapman-Richards function. For spruce and beech, generalized additive models of the SI against simple climate variables lead to stable and plausible model behavior.

Conclusion

The introduced methodology permits a harmonized quantification of forest site productivity by static site indices. Predicting productivity in dependence on climate illustrates the benefits of combined datasets.

     

Impact of bark and foliage harvesting on fruit production of the multipurpose tree <Emphasis Type="Italic">Afzelia africana</Emphasis> in Burkina Faso (West Africa)

In sub-Saharan Africa, extraction for daily livelihood needs often results in uncontrolled exploitation of bark and leaves of valuable medicinal and fodder trees. However, overharvesting of bark and foliage can reduce fruit production and threaten reproduction. This study evaluates the impact of combined bark and foliage harvesting on the performance of fruit production of Afzelia africana in Burkina Faso. We compared fruit and seed production at different harvesting intensities. Data on fruit yields were collected by stratified random sampling of 91 trees with no, low, severe, and very severe harvesting intensities. The fruit production varied with harvesting intensity, tree size and number of branches. Fruit and seed quantity and quality decreased with increasing harvesting intensity. However, no significant difference was detected between trees without and trees under low harvesting. Trees of all size classes under very severe harvesting intensity had no fruits. Under low harvesting impact, large trees had twice as many fruits as the control, whereas fruits were reduced by half to 95 % for the small trees. High harvesting intensity is an unsustainable practice that should be completely prohibited in order to ensure longterm persistence of Afzelia africana. Low harvesting intensity should be allowed, but only on large reproductive individuals.     

Low expression of nicotinic acetylcholine receptor subunit Mdα2 in neonicotinoid‐resistant strains of Musca domestica L.

BACKGROUND: Neonicotinoid action as well as resistance involves interaction with nicotinic acetylcholine receptors (nAChRs). In the housefly, neonicotinoid resistance also involves cytochrome P450, as indicated by bioassay with synergist as well as altered expression. In bioassay, synergism was only partial and indicated possible target‐site resistance. The nAChR α2 subunit is important in neonicotinoid toxicity to insects, and gene expression of the Mdα2 subunit was investigated in field populations and laboratory strains of neonicotinoid‐resistant and insecticide‐susceptible houseflies, Musca domestica L. The genomic sequence covering exon III–VII of Mdα2 was analysed for mutations. RESULTS: Gene expression profiling of Mdα2 revealed notable differences between neonicotinoid‐resistant and insecticide‐susceptible houseflies. On average, the neonicotinoid‐resistant field population 766b and the imidacloprid selected strain 791imi had 60% lower copy numbers of Mdα2 compared with the susceptible reference strain. Sequencing of exon III–VII of the Mdα2, encoding acetylcholine binding‐site regions and three out of four transmembrane domains, did not reveal any mutations explaining the increased neonicotinoid tolerance in the strains examined. CONCLUSION: Previous discoveries and the results of this study suggest that the neonicotinoid resistance mechanism in Danish houseflies involves both cytochrome P450 monooxygenase‐mediated detoxification and reduced expression of the nAChR subunit α2. Copyright © 2010 Society of Chemical Industry     

Protocol: using virus-induced gene silencing to study the arbuscular mycorrhizal symbiosis in <Emphasis Type="Italic">Pisum sativum</Emphasis>

Virus-induced gene silencing (VIGS) is an alternative reverse genetics tool for silencing of genes in some plants, which are difficult to transform. The pea early-browning virus (PEBV) has been developed as a VIGS vector and used in pea for functional analysis of several genes. However, the available PEBV-VIGS protocols are inadequate for studying genes involved in the symbiosis with arbuscular mycorrhizal fungi (AMF).