Effect of decreasing dietary crude protein in fattening calves on the emission of ammonia and greenhouse gases from manure stored under aerobic and anaerobic conditions

Dietary strategies can potentially help to reduce nitrogen (N) emissions and decrease the environmental impact of beef production. This study aimed to evaluate the effects of dietary crude protein (CP) concentration on animal performance, N excretion, and manure N volatilisation of finishing Holstein animals. In a first study, 105 Holstein bulls (BW 344 ± 2.6 kg; age 252 ± 0.9 days) were allocated to eight pens to evaluate the effect of two treatments (medium (M) and low (L), which contained CP 14.5% and 12% on a DM basis, respectively) on performance, and results confirmed that dietary CP decrease did not impair animal growth. In a second study, N excretion study, 24 Holstein heifers (BW 310 ± 5.3 kg; age 251 ± 1.4 days) were distributed randomly depending on the initial BW to three treatments (high (H), M, and L, which contained CP 17%, 14.5% and 12% on a DM basis, respectively). Based on N excretion, urinary N excretion was greater (P < 0.001) in H than in M and L diets, but no differences in faecal N excretion were observed among treatments. A third study with in vitro assays under aerobic and anaerobic conditions was designed to analyse gaseous emissions (volatilisation of N and carbon, C) during the storage stage of manure. Manure, faecal and urine samples, mixed at a ratio of 1:1 (wet weight), were collected during the N excretion study (manure-H, manure-M, manure-L). Under aerobic conditions, manure-M and manure-L showed a delay of 4–5 days in manure ammonia emission compared with manure-H (P < 0.01). Total N content was lower (P < 0.01) in manure-L compared with manure-M and manure-H, but N volatilisation (percentage relative to initial N) in manure-L and manure-M was greater (P < 0.01) than in manure-H. In contrast, the anaerobic N volatilisation was 20 times greater in manure-M and 10 times greater in manure-H compared with manure-L. Under aerobic and anaerobic conditions, the emission of C, as C-CO₂ and C-CH₄, was greater in manure-L than in manure-H and manure-M. Therefore, the decrease of dietary CP concentration from 17% to 14.5% and 12% is an efficient strategy to reduce urinary N excretion by 40%, without impairing performance, and also to reduce manure N losses through ammonia volatilisation under anaerobic conditions. However, a dietary CP content of 14.5% resulted in less environmental impact than a CP content of 12.8% when also considering manure emissions under aerobic or anaerobic conditions.     

Down-regulation of the non-neuronal acetylcholine synthesis and release machinery in acute allergic airway inflammation of rat and mouse

Acetylcholine (ACh), derived both from nerve fibres and from non-neuronal sources such as epithelial cells, is a major regulator of airway function. There is evidence that dysfunction of the neuronal cholinergic system is involved in the pathogenesis of asthma. Here, we asked whether the pulmonary non-neuronal ACh-synthesis and release machinery is altered in a rat and a mouse model of allergic airway disease. Animals were sensitized against ovalbumin, challenged by allergen inhalation, and sacrificed 24 or 48 h later. Targets of investigation were the high-affinity choline transporter-1 (CHT1), that mediates cellular uptake of choline, the ACh-synthesizing enzyme choline acetyltransferase (ChAT), the vesicular ACh transporter (VAChT), and the polyspecific organic cation transporters (OCT1-3), which are able to translocate choline and ACh across the plasma membrane. With cell-type specific distribution patterns, immunohistochemistry identified these proteins in airway epithelial cells and alveolar macrophages. Real-time RT-PCR revealed significant decreases in ChAT-, CHT1-, VAChT-, OCT-mRNA in the lung of sensitized and allergen challenged animals. These data were supported by immunohistochemistry, demonstrating reduced labeling intensity of airway epithelial cells. ChAT-, CHT1-, VAChT-, and OCT1-mRNA were also significantly reduced in cells recovered by bronchoalveolar lavage from sensitized and challenged rats. In conclusion, the pulmonary non-neuronal cholinergic system is down-regulated in acute allergic airway inflammation. In view of the role of ACh in maintenance of cell-cell-contacts, stimulation of fluid-secretion and of ciliary beat frequency, this down-regulation may contribute to epithelial shedding and ciliated cell dysfunction that occur in this pathological condition.     

A murine model of infection with Rickettsia prowazekii: implications for pathogenesis of epidemic typhus

Epidemic typhus remains a major disease threat, furthermore, its etiologic agent, Rickettsia prowazekii, is classified as a bioterrorism agent. We describe here a murine model of epidemic typhus that reproduced some features of the human disease. When BALB/c mice were inoculated intravenously with R. prowazekii (Breinl strain), they survived but did not clear R. prowazekii infection. Immunohistological analysis of tissues and quantitative PCR showed that R. prowazekii was present in blood, liver, lungs and brain 1 day after infection and persisted for at least 9 days. Importantly, infected mice developed interstitial pneumonia, with consolidation of the alveoli, hemorrhages in lungs, multifocal granulomas in liver, and hemorrhages in brain, as seen in humans. Circulating antibodies directed against R. prowazekii were detected at day 4 post-infection and steadily increased for up to 21 days, demonstrating that R. prowazekii lesions were independent of humoral immune response. R. prowazekii-induced lesions were associated with inflammatory response, as demonstrated by elevated levels of inflammatory cytokines including interferon-gamma, tumor necrosis factor and the CC chemokine RANTES in the lesions. We concluded that the BALB/c mouse strain provides a useful model for studying the pathogenic mechanisms of epidemic typhus and its control by the immune system.     

Characterization of a metabotropic glutamate receptor in the honeybee (<Emphasis Type="Italic">Apis mellifera</Emphasis>): implications for memory formation

G-protein-coupled metabotropic glutamate receptors (GPC mGluRs) are important constituents of glutamatergic synapses where they contribute to synaptic plasticity and development. Here we characterised a member of this family in the honeybee. We show that the honeybee genome encodes a genuine mGluR (AmGluRA) that is expressed at low to medium levels in both pupal and adult brains. Analysis of honeybee protein sequence places it within the type 3 GPCR family, which includes mGlu receptors, GABA-B receptors, calcium-sensing receptors, and pheromone receptors. Phylogenetic comparisons combined with pharmacological evaluation in HEK 293 cells transiently expressing AmGluRA show that the honeybee protein belongs to the group II mGluRs. With respect to learning and memory AmGluRA appears to be required for memory formation. Both agonists and antagonists selective against the group II mGluRs impair long-term (24 h) associative olfactory memory formation when applied 1 h before training, but have no effect when injected post-training or pre-testing. Our results strengthen the notion that glutamate is a key neurotransmitter in memory processes in the honeybee.     

Application of phage display to high throughput antibody generation and characterization

We have created a high quality phage display library containing over 10¹⁰ human antibodies and describe its use in the generation of antibodies on an unprecedented scale. We have selected, screened and sequenced over 38,000 recombinant antibodies to 292 antigens, yielding over 7,200 unique clones. 4,400 antibodies were characterized by specificity testing and detailed sequence analysis and the data/clones are available online. Sensitive detection was demonstrated in a bead based flow cytometry assay. Furthermore, positive staining by immunohistochemistry on tissue microarrays was found for 37% (143/381) of antibodies. Thus, we have demonstrated the potential of and illuminated the issues associated with genome-wide monoclonal antibody generation.     

New Pedal Remains of Megaladapis and Their Functional Significance. Am J Phys Anthropol 100:115–139

Erratum: Wunderlich RE, Simons EL, Jungers WL. 1996. New Pedal Remains of Megaladapis and Their Functional Significance. Am J Phys Anthropol 100:115–139.     

Specific detection of Phialophora gregata and Plectosporium tabacinum in infected soybean plants using polymerase chain reaction

The fungal deuteromycetes Phialophora gregata and Plectosporium tabacinum are associated with soybean plants. P. gregata causes brown stem rot (BSR) of soybean, whereas P. tabacinum is a frequent cohabitant of soybean stems. The role of P. tabacinum in soybean growth and in the development of BSR is not known. Traditional methods of isolating and differentiating these two fungi require up to 3 weeks to complete. In order to effectively study the interactions among P. gregata, P. tabacinum , and the soybean plant, we developed specific primers for P. tabacinum based on its rDNA internal transcribed spacer sequences. In combination with specific primers developed previously for P. gregata , the specific primer pairs were used successfully in polymerase chain reactions to detect the targeted fungi in both artificially inoculated and naturally infected soybean plants. Using this technique, we examined 130 soybean plants collected from natural field environments for the presence or absence of P. gregata and P. tabacinum . Statistical analyses of the results showed that the frequency of co-occurrence of P. gregata and P. tabacinum in soybean plants was significantly less than expected if the two fungi would occur independently, suggesting that one of the fungi may be inhibitory to the other fungus.