Effect of dietary nitrogen content on the urine metabolite profile of dairy cows assessed by nuclear magnetic resonance (NMR)-based metabolomics

NMR-based metabolomics was applied on urine samples from 32 cows that were fed four levels of crude protein (124, 135, 151, and 166 g/kg DM, respectively) in a crossover design with the aim of identifying urinary metabolites related to nitrogen intake and nitrogen efficiency. Principal component analysis (PCA) on selected regions of the obtained (1)H NMR spectra revealed an effect of crude protein intake on NMR signals in the 0.5-3.0 and 5.0-10.0 ppm regions. Partial least-squares (PLS) regressions confirmed a correlation between the NMR metabolite profile and both nitrogen intake and efficiency. The NMR signals that correlated with nitrogen intake and efficiency included urea, hippurate, phenylacetylglutamine, and p-cresol sulfate, which all contributed to the prediction of nitrogen intake and efficiency. Thus, it was not possible to identify a single metabolite that could be used as a marker to predict nitrogen efficiency, and it can be concluded that a wide-ranging urinary metabolite profile is needed to evaluate nitrogen efficiency in ruminants.     

Uterus duplex bicollis,Vagina simplex in laboratory Guinea pigs (Cavia porcellus), rats (Rattus norvegicus forma domestica) and mice (Mus musculus forma domestica)

The female genital tract originates from the Müllerian ducts during embryological development. Fusion of the ducts occurs in different segments depending on the animal species, resulting in a variational number of the respective organ. Current literature on genital tract morphology of laboratory rodents is controversial. Therefore, the present study aimed at determining an anatomically correct definition of the uterus in laboratory guinea pigs, mice and rats. In all three rodent species, we found two separate cervical canals that communicate with an individual uterus via discrete ostia uteri interna. The correct anatomical definition should therefore be uterus duplex bicollis, vagina simplex.     

Origin of the high-frequency resonances in 1H NMR spectra of muscle tissue: an in vitro slow magic-angle spinning study

High-resolution slow magic-angle spinning (150 Hz) 1H PASS NMR spectroscopy is performed on intact excised rat m. tibialis anterior. Untreated muscles and muscles in vitro incubated in Krebs-Ringers buffer based on deuterium oxide are investigated. In the high-frequency region of the 1H NMR spectra, resonances from H4 (approximately 7.1-7.2 ppm) and H2 (approximately 8.2-8.5 ppm) in histidine are observed. In addition, a resonance appears at 6.7 ppm for the untreated muscles. However, this resonance is absent in muscles following incubation in deuterium oxide. On the basis of its behavior in deuterium oxide combined with supplementary measurements for creatine solutions, the 6.7 ppm resonance is ascribed to the amino protons in creatine. Moreover, the present study demonstrates that the observation of the 6.7 ppm resonance depends on pH, which explains earlier reports stating its occasional appearance. Finally, measurements on solutions of ATP/AMP and histidine indicate that both ATP/AMP and histidine contribute to the resonances at approximately 8.2-8.5 ppm in the 1H NMR spectra of muscle tissue.     

Structural analogues of homoeriodictyol as flavor modifiers. Part III: short chain gingerdione derivatives

In order to find new flavor modifiers, various short chain gingerdione derivatives were synthesized as structural analogues of the known bitter masker homoeriodictyol and evaluated by a sensory panel for masking and sweetness enhancing activities. 1-(4-Hydroxy-3-methoxyphenyl)hexa-3,5-dione ([2]-gingerdione) and the homologue 1-(4-hydroxy-3-methoxyphenyl)hepta-3,5-dione ([3]-gingerdione) at concentration ranges 50-500 mg kg (-1) showed the most promising masking activity of 20-30% against bitterness of a 500 mg kg (-1) aqueous caffeine solution. Additionally, both compounds were able to reduce the bitterness of a 5 mg kg (-1) quinine solution by about 20%; however, the bitter tastes of salicine, the model peptide H-Leu-Trp-OH, and KCl solutions were not reduced. Whereas for bitter masking activity a vanillyl moiety seems to be important, some of the tested isovanillyl isomers showed an interesting sweet enhancing effect without exhibiting a significant intrinsic sweetness. The isomer 1-(3-hydroxy-4-methoxyphenyl)hexa-3,5-dione ([2]-isogingerdione) at 100 mg kg (-1) caused a significant and synergistic increase of 27% of sweet taste of a 5% sucrose solution.     

Pathology of an ulcerative dermatitis in Belgian Landrace sows

Chronic ulcers of the skin were observed in three Belgian Landrace sows. Lesions were located on ears, limbs, and in the mammary region and were resistant to treatment that included corticosteroid therapy. Major histologic changes were located at the dermo-epidermal junction. Ulcers were deep, and the adjacent epidermis had marked hyperkeratosis, acanthosis, and intracellular edema. Dermatitis was prominent, essentially located in the superficial dermis. By electron microscopy, basal lamina appeared disrupted. Dermo-epidermal separation occurred beneath the basal lamina. Collagen was morphologically normal. Desmosomes, hemidesmosomes, and anchorage fibers were present in areas adjacent to lesions. Ulcerative dermatitis of sows is morphologically similar to pemphigus, pemphigoid, systemic lupus erythematosus, epidermolysis bullosa simplex, erythema multiforme, toxic epidermal necrolysis, and drug eruption. However, significant differences exist between ulcerative dermatitis and these conditions.     

Impact of bovine urine deposition on soil microbial activity,biomass, and community structure

Nitrogen (N) from urine excreted by grazing animals can be transformed into N compounds that have detrimental effects on the environment. These include nitrate, which can cause eutrophication of waterways, and nitrous oxide, which is a greenhouse gas. Soil microbes mediate all of these N transformations, but the impact of urine on microbes and how initial soil conditions and urine chemical composition alter their responses to urine are not well understood. This study aimed to determine how soil inorganic N pools, nitrous oxide fluxes, soil microbial activity, biomass, and the community structure of bacteria containing amoA (nitrifiers), nirK, and nirS (denitrifiers) genes responded to the addition of urine over time. Bovine urine containing either a high (15.0 g K⁺ l^?1) or low salt content (10.4 g K⁺ l^?1) was added to soil cores at either low or high moisture content (hereafter termed dry and wet soil respectively; 35% or 70% water-filled pore space after the addition of urine). Changes in soil conditions, inorganic N pools, nitrous oxide fluxes, and the soil microbial community were then measured 1, 3, 8, 15, 29 and 44 days after urine addition. Urine addition increased soil ammonium concentrations by up to 2 mg g d.w.^?1, soil pH by up to 2.7 units, and electrical conductivity (EC) by 1.0 and 1.6 dS m^?1 in the low and high salt urine treatments respectively. In response, nitrate accumulation and nitrous oxide fluxes were lower in dry compared to wet urine-amended soils and slightly lower in high compared to low salt urine-amended soils. Nitrite concentrations were elevated (>3 μg g d.w.^?1) for at least 15 days after urine addition in wet urine-amended soils, but were only this high in the dry urine-amended soils for 1 day after the addition of urine. Microbial biomass was reduced by up to half in the wet urine-amended soils, but was largely unaffected in the dry urine-amended soils. Urine addition affected the community structure of ammonia-oxidising and nitrite-reducing bacteria; this response was also stronger and more persistent in wet than in dry urine-amended soils. Overall, the changes in soil conditions caused by the addition of urine interacted to influence microbial responses, indicating that the effect of urine on soil microbes is likely to be context-dependent.     

Functional and ultrastructural changes in neutrophils from mares and foals experimentally inoculated with a respiratory tract strain of equine herpesvirus-1

Neutrophils isolated from venous blood of adult and foal ponies inoculated with equine herpesvirus-1 were evaluated by in vitro function tests and by electron microscopy. Foals had fever and severe neutropenia 24 hours after inoculation; increased neutrophil random migration under agarose and decreased antibody-dependent cell-mediated cytotoxicity were significant at 24 hours, but values had returned to preinoculation levels by 72 hours. Mares had fever and leukopenia of less severity, increases in neutrophil migration, and longer persistence of primary granule release than were seen in foals. Reduced migration and degranulation, and a decrease in antibody-dependent cell-mediated cytotoxicity seen with neutrophils from foals, as compared with mares, may relate to the high susceptibility of foals to equine herpesvirus-1 infection.     

Pathogenicity of equine herpesvirus 1 subtype 2 for foals and adult pony mares

Three pony mares and 4 pony foals were inoculated with a subtype 2 strain of equine herpesvirus 1. Foals had periods of fever 12 h and 2.5 days after inoculation and leukopenia, involving both neutrophils and lymphocytes, followed by leukocytosis. Mares had transient fever and leukopenia 24 hours after inoculation that were less severe than in foals. An increase in circulating virus-neutralizing antibody was seen in 2 of 3 inoculated mares, but not in foals. Attempts to isolate virus from blood were unsuccessful. These studies show that equine herpesvirus 1 subtype 2 is a mild pathogen for ponies and infection may result in inapparent clinical disease.     

Intravascular macrophages in lungs of pigs infected with Haemophilus pleuropneumoniae

Pigs were inoculated intratracheally with a virulent or an avirulent isolate of Haemophilus pleuropneumoniae serotype 5 and sacrificed during the first 24 hours post-inoculation. Intravascular macrophages were examined by electron microscopic and morphometric techniques. Samples of lung were taken from regions with no macroscopic lesions (Zone 0), 2.5 to 3.0 cm from lesions (Zone 1), and from the immediate edge of lesions (Zone 2). Those pigs inoculated with the avirulent isolate did not develop lesions. Pigs given the virulent isolate consistently developed necrohemorrhagic lesions in the dorsolateral aspect of the caudal and middle lung lobes. Relative volumes of intravascular macrophages in Zones 1 and 2 increased with increased time post-inoculation; in pigs given the avirulent isolate, intravascular macrophage volume decreased with increased time post-inoculation. Cytoplasmic volume to nuclear volume ratios for macrophages in Zone 2 from pigs with necrohemorrhagic lesions progressively increased with increased time post-inoculation. Enlarged intravascular macrophages had large nuclei, prominent nucleoli, and abundant cytoplasm. Increased cytoplasmic volume was the result of increased numbers of lysosomes, phagosomes, rough and smooth endoplasmic reticulum, and large Golgi complexes. Pigs inoculated with the virulent bacteria had IV macrophages with large phagosomes that contained necrotic cell debris and fibrin. Macrophages with phagosomes were more frequent in the 6-, 9-, and 24-hour sample periods of pigs with lesions than in any other group. Intercellular adhesion plaques (ICAP) were present between IV macrophages and subjacent endothelial cells. ICAP's increased in length with increased time post-inoculation in Zones 1 and 2 from pigs with necrohemorrhagic lesions. In later sample periods, multiple closely associated and interlacing IV macrophages formed a discontinuous layer over endothelial cells in Zone 2 samples near necrohemorrhagic lesions. These results suggest that the intravascular macrophage population changes from immature macrophages to mature macrophages or immature epithelioid cells within 24 hours after inhalation of a virulent Haemophilus pleuropneumoniae. Furthermore, intravascular macrophages likely function to clear cellular and acellular debris from the blood in pneumonic conditions.     

Responses of equine neutrophils to contagious equine metritis organism and its lipopolysaccharides

Morphology and function of equine neutrophils were evaluated after combination with contagious equine metritis organism (CEMO) or 1 of 2 CEMO lipopolysaccharides (LPS). The 2 LPS (LPS-a; LPS-p) isolated from the CEMO contained 14- and 16-carbon fatty acids, ketodeoxyoctanate, hexose, and heptose, but were morphologically distinct. Neutrophils exposed to LPS had fewer granules, whereas those exposed to CEMO had more granules than did the controls (phosphate-buffered saline solution). Neutrophil iodination was significantly increased with 10 and 25 micrograms of LPS-a, but not significantly altered by LPS-p or CEMO. Staphylococcus aureus ingestion was not influenced by CEMO and was mildly decreased by LPS-a. These results indicate that CEMO may have at least 2 functionally and morphologically distinct, but chemically similar, LPS and that 1 of these LPS (LPS-a) may enhance neutrophil killing by stimulating neutrophil iodinating mechanisms.