Nitrogen metabolism and fertility in cattle: I. Adaptive changes in intake and metabolism to diets differing in their rate of energy and nitrogen release in the rumen

The ruminal degradability, intake, and metabolism of diets differing in their relative rate of energy and nitrogen release in the rumen were characterized prior to their use in a study of the effects of high peripheral levels of ammonia on reproductive function in cattle. In a 2 x 2 factorial experiment, replicated four times, 16 heifers were offered isocaloric and isonitrogenous diets containing two sources of fermentable carbohydrate, fiber (slow energy release, SE) or starch (fast energy release, FE), and two rates of nitrogen release, which were either synchronous (S) or asynchronous (A) to that of energy release. Throughout the experiment, the amount of feed offered was held constant, at a level equivalent to 1.5 x maintenance. Four ruminally fistulated sheep were used to determine the in situ degradability of these diets. The 16 heifers were bled before feeding at 0800 and at 0900, 1000, 1100, 1200, 1400, and 1600 on d 0 (introduction to dietary treatments) and on d 4, 7, 11, 14, 21, and 28. Diet refusals were recorded at hourly intervals after feeding. The rapidly degradable nitrogen fraction of the SE:A and FE:A diets was greater than that of the SE:S and FE:S diets. Postprandial jugular plasma ammonia levels rose to a peak of around 300 micromol/L in heifers offered the SE:A and FE:A diets but did not rise in heifers offered the SE:S and FE:S diets. All feed offered was consumed within 1 h on diets SE:S and FE:S throughout the experiment. The proportion of feed consumed within 1 h of feeding declined from 100% on d 0 to around 70 and 56% by d 21 for heifers given the SE:A and FE:A diets, respectively. Peak postprandial plasma ammonia levels were accordingly lower, at around 160 micromol/L. Plasma urea levels averaged 7 mmol/L and were unaffected by dietary treatment. High plasma ammonia levels were associated with a suppression in the normal postprandial rise in insulin. There was no significant metabolic adaptation to high-ammonia-generating diets, and heifers given these diets modified their pattern of intake in an apparent attempt to avoid excessively high levels of plasma ammonia.     

Evidence for surfactant contributing to the gastric mucosal barrier of the horse

This study was undertaken to determine the hydrophobicity of the luminal surface of the equine stomach and to elucidate the ultrastructure of the lining imparting that property. Gastric and duodenal mucosal samples from 5 horses were collected immediately after euthanasia and subjected to surface contact angle measurement using a goniometer. Gastric mucosal samples from 4 horses and a foal were examined by electron microscopy following a fixation procedure known to preserve phospholipids and oligolamellar structures. Contact angles for the equine gastric glandular mucosal surface (mean +/- s.e. 78.0 +/- 11.0 degrees) were greater than for the duodenum (33.4 +/- 8.7 degrees), (P = 0.003). The contact angles for gastric squamous tissue (50.4 +/- 4.5 degrees) tended to be greater than for duodenum (P = 0.15). Electron microscopy revealed the existence of surfactant as abundant osmiophilic phospholipid material within both squamous and glandular gastric mucosae. These results indicate the hydrophobic nature of the equine gastric mucosae. We propose that the water-repellent nature of the stomach contributes to the 'gastric mucosal barrier' and is imparted by surface-active phospholipid adsorbed to the surface. Phospholipids may also be utilised as a physical barrier to back-diffusion of acid by lining intracellular canaliculi and oxyntic ducts where other defence mechanisms are absent.     

Effect of intramammary infusion of tumour necrosis factor-alpha on milk protein composition and induction of acute-phase protein in the lactating cow

The present study was designed to evaluate the effects of tumour necrosis factor-alpha (TNF-alpha) on lactating bovine mammary functions such as milk protein secretion and the integrity of the milk-blood barrier. The effect on the induction of the systemic inflammatory response was also examined using concentrations of serum haptoglobin (Hp), a major inflammatory acute-phase protein, as an index. One hundred micrograms per mammary gland of recombinant bovine (rBo) TNF-alpha or placebo saline was individually infused into a rear mammary gland of each of four lactating cows, and milk and blood samples were collected before and 4, 8, 24, 32, 48, 96 and 168 h after infusion. In the rBoTNF-alpha-infused gland, increases of somatic cell counts were observed at 4-48 h. Although concentrations of total milk protein were not changed, compositions of milk proteins varied following rBoTNF-alpha infusion. Concentrations of caseins, alpha-lactalbumin and beta-lactoglobulin were significantly decreased at 4 and 8 h. Lactoferrin concentrations were significantly increased at 4 h. Significant infiltrations of serum albumin, immunoglobulin G1 (IgG1) and IgG2 were observed at 4 and 8 h. Elevations of the serum concentration of Hp were detected at 8-32 h, but were very small in comparison with those reported in inflammatory diseases. Changes in rectal temperature and white blood cell counts were not significant. These results show that single rBoTNF-alpha infusion into the lactating mammary gland suppresses the lactogenic function of the gland and influences the function of the milk-blood barrier, with little effect on the generalized inflammatory response.     

Prevalence and intensity of Otodectes cynotis in kittens from Thessaloniki area, Greece

From May 2007 to May 2008 we have examined by otoscopy a total number of 214 cats, aged between 0 and 6 months, brought in for their first veterinary examination to a private veterinary clinic. All cats were of urban origin. In all positive cats we performed a washing of the ear using warm paraffin oil in order to determine intensity of infection. Statistical analysis was performed using the Chi-square test. A total number of 30 cats were found to be infected with Otodectes cynotis (prevalence 14.02%). Prevalence of infection was significantly (p < 0.05) higher in cats aged between 3 and 6 months (17.58%) than in cats aged below 3 months (11.38). Intensity of infection ranged between 7 and 85 (mean intensity 35.60) mites per infected cat. The same age group of cats between 3 and 6 months had significantly (p < 0.05) higher mean intensity (47.19) compared to cats aged under 3 months (22.36). No statistical significance was found between males and females for neither prevalence nor intensity. Differences and similarities with other studies are discussed.     

Cytokine profiles, apoptosis and pathology of experimental Pasteurella multocida serotype A1 infection in mice

Mice were experimentally infected with Pasteurella multocida serotype A1 to study the cytokine profiles, host cell apoptosis and sequential pathology at different hours of post-infection. Infected mice were dull, anorectic and depressed. A transient leukocytopenia followed by progressive leukocytosis was observed in the course of infection. Serum cytokine profiles showed significantly (P < 0.01) higher amount of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6 and mouse KC) in the infected mice when compared to control mice. The circulating lymphocytes were apoptotic on annexin V staining. Apoptotic nuclei were detected in splenocytes, hepatocytes and infiltrating leukocytes of the lungs on TUNEL staining. The lungs were grossly congested and hemorrhagic, and showed infiltration with polymorphonuclear cells at early and mononuclear cells in the late hours of infection. Alveolar epithelia, inter-alveolar septa and capillary endothelium of the lungs showed ultrastructural changes. Liver had degenerative changes in histological and ultrathin sections.     

Evaluation of oxidative stress in hunting dogs during exercise

Exercise has been shown to increase the production of reactive oxygen species (ROS) to a point that can exceed antioxidant defenses, to cause oxidative stress. The aim of our trials was to evaluate oxidative stress and recovery times in trained dogs during two different hunting exercises, with reactive oxygen metabolites-derivatives (d-ROMs) and biological antioxidant potential (BAP) tests. A group of nine privately owned Italian hounds were included. A 20-min aerobic exercise and a 4-h aerobic exercise, after 30 days of rest, were performed by the dogs. Our results show an oxidative stress after exercise due to both the high concentration of oxidants (d-ROMs) and the low level of antioxidant power (BAP). Besides, the recovery time is faster after the 4-h aerobic exercise than the 20-min aerobic exercise. Oxidative stress monitoring during dogs exercise could become an interesting aid to establish ideal adaptation to training.     

Effect of Enterococcus faecium AL41 and Thymus vulgaris essential oil on small intestine integrity and antioxidative status of laying hens

We investigated the effect of Enterococcus faecium on phagocytic activity, antioxidative status in vivo and the effect of E. faecium and 0.4% concentration of Thymus vulgaris essential oil (EO) on the duodenal tissue integrity in vitro in laying hens. The birds were fed the same standard diets and were divided into four groups. E. faecium was added to the drinking water for the second and fourth groups. EO was added to special chambers for measuring trans-epithelial electrical resistance (TEER) for the third and fourth groups only. TEER was lower in groups where EO was added, but in the group with E. faecium TEER was not changed significantly. Our results show that EO at 0.4% concentration may negatively affect intestine integrity, and the probiotic strain E. faecium AL41 is able to eliminate this effect and can strengthen non-specific immunity. To confirm our findings further histopathological investigations of intestinal tissue are needed.     

Body composition and tissue accretion rates of barrows fed corn-soybean meal diets or low-protein, amino acid-supplemented diets at different feeding levels

Two experiments, each with 39 high-lean-gain potential barrows, were conducted to evaluate the organ weights, body chemical composition, and tissue accretion rates of pigs fed corn-soybean meal diets (CONTROL) and low-protein diets supplemented with crystalline lysine, threonine, tryptophan, and methionine either on an ideal protein basis (IDEAL) or in a pattern similar to that of the control diet (AACON). Amino acids were added on a true ileally digestible basis. The initial and final BW were, respectively, 31.5 and 82.3 kg in Exp. 1 and 32.7 and 57.1 kg in Exp. 2, and pigs were fed for 55 and 27 d in Exp. 1 and 2, respectively. In Exp. 1, the CONTROL and IDEAL diets were offered on an ad libitum basis, or by feeding 90 or 80% of ad libitum intake. In Exp. 2, the CONTROL, IDEAL, and AACON diets were offered on an ad libitum basis, or by feeding 80% of the ad libitum intake. Three pigs were killed at the start of the experiments and three from each treatment were killed at the end of each experiment to determine body chemical composition. In both trials, the whole-body protein concentration (g/kg) and the accretion rates of protein (g/d) were greater (P < 0.05) for pigs fed the CONTROL than for pigs fed the IDEAL and AACON diets. In Exp. 1, pigs fed the CONTROL diet had a trend (P < 0.10) for greater water and lower lipid concentration and had greater (P < 0.05) water and ash accretion rates. Whole-body protein concentration was greatest (P < 0.05) in pigs fed at 80% of ad libitum, but protein, water, and ash accretion rates were greatest (P < 0.05) in pigs allowed ad libitum access to feed. In summary, pigs fed the IDEAL and the AACON diets had less protein in the body and lower protein accretion rates than pigs fed the CONTROL diet. It seems that reductions in protein deposition in pigs fed the IDEAL and AACON diets may have been due to a deficiency of one or more essential amino acids or possibly to increases in the NE for metabolic processes leading to increases in adipose tissue deposition.     

Relative bioactivity of dietary RRR- and all-rac-alpha-tocopheryl acetates in swine assessed with deuterium-labeled vitamin E

This study evaluated the relative bioactivities of natural and synthetic stereoisomers of alpha-tocopherol in swine. Deuterium-labeled vitamin E (150 mg each of d3-RRR- [natural] and d6-all-rac- [synthetic] alpha-tocopheryl acetates) was administered orally to adult female pigs (n = 3) with the morning feed. Blood samples were obtained at 0, 3, 6, 9, 12, 36, 48, and 72 h after the dose. The time of maximum plasma d3-alpha-tocopherol concentration (0.486 microg/mL) occurred at 12 h, and d6-alpha-tocopherol peaked earlier (at 9 h) and at a lower (P < 0.05) concentration (0.288 microg/mL). The d3-/d6-alpha-tocopherol ratio increased from 1.35 (SD = 0.73) at 3 h after dosing to 2.0 (SD = 0.14) at 72 h (P = 0.03). The plasma disappearance rates of d3- and d6-alpha-tocopherols (post-maximum concentrations) were similar and were estimated to be 0.013 microg/mL per hour. In summary, swine discriminated between RRR- and all-rac-alpha-tocopherols, which resulted in an approximately twofold higher plasma alpha-tocopherol concentration arising from the RRR-form. This 2:1 ratio of RRR- to all-rac- is higher than the currently accepted USP definition of RRR-:all-rac- of 1.36:1.00.     

A comparison of enzymatic and molecular approaches to characterize the cellulolytic microbial ecosystems of the rumen and the cecum

We used RNA probes and enzyme activities to compare the cellulolytic microbial ecosystems of the rumen and the cecum. Four rumen- and cecum-cannulated wethers were fed a diet of barley plus hay (60:40). Digesta samples were collected 1 h before feeding and 3, 6, and 9 h after feeding for measurements on microbial populations, and 1 h before feeding and 3 and 6 h after feeding for digestion measurements, pH, and VFA. Polysaccharidase and glycosidase specific activities of solid-adherent microorganisms were measured respectively by the amount of reducing sugars released from xylan or avicel or p-nitrophenol from the p-nitrophenol derivatives of xylose and glucose. The distribution and amounts of the three main cellulolytic bacterial species (Fibrobacter succinogenes, Ruminococcus albus, and Ruminococcus flavefaciens) were determined by dot-blot hybridization using specific 16SrRNA-targeting probes. Enzyme activities were higher in the rumen than in the cecum and before feeding than at 3 h after feeding. The sum of the three cellulolytic bacterial species represented, on average, 4.5% of the total bacterial RNA in the two compartments and did not vary with sampling time. The cellulolytic bacterial community structure was different in the two compartments, with F. succinogenes as the main species in the rumen and R. flavefaciens in the cecum. The lower cellulolytic activity in the cecum than in the rumen could not be ascribed to any difference in the structure of the cellulolytic bacterial community between these two compartments, and other hypotheses related to digestion are proposed.