The influence of glyphosate on the microbiota and production of botulinum neurotoxin during ruminal fermentation.

The aim of the present study is to investigate the impact of glyphosate on the microbiota and on the botulinum neurotoxin (BoNT) expression during in vitro ruminal fermentation. This study was conducted using two DAISY(II)-incubators with four ventilated incubation vessels filled with rumen fluid of a 4-year-old non-lactating Holstein-Friesian cow. Two hundred milliliter rumen fluid and 800 ml buffer solution were used with six filter bags containing 500 mg concentrated feed or crude fiber-enriched diet. Final concentrations of 0, 1, 10, and 100 µg/ml of glyphosate in the diluted rumen fluids were added and incubated under CO2-aerated conditions for 48 h. The protozoal population was analyzed microscopically and the ruminal flora was characterized using the fluorescence in situ hybridization technique. Clostridium botulinum and BoNT were quantified using most probable number and ELISA, respectively. Results showed that glyphosate had an inhibitory effect on select groups of the ruminal microbiota, but increased the population of pathogenic species. The BoNT was produced during incubation when inoculum was treated with high doses of glyphosate. In conclusion, glyphosate causes dysbiosis which favors the production of BoNT in the rumen. The global regulations restrictions for the use of glyphosate should be re-evaluated.

Protein Evaluation of Feedstuffs for Horses.

The German Society of Nutrition Physiology has proposed a new protein evaluation system for horse feeds to estimate pre-cecally digestible crude protein (pcdCP) and amino acids (pcdAA) from chemical properties. A total of 71 feeds for horses were chemically tested and evaluated according to the new protein evaluation system. A feeding trial with eight horses tested whether differences in estimated pcdAA and neutral detergent soluble CP (NDSCP) in the diet were reflected by post-prandial (ppr) kinetics of plasma lysine (Lys) by feeding a complementary feed (control = CTRL) with 1.02 g Lys/100 kg body weight (BW) as well as three diets with 3.02 g Lys/100 kg BW, as follows: (i) CTRL with synthetic AA (CTRL + synAA); (ii) CTRL with soybean meal (CTRL + SBM); and (iii) lucerne pellets (LUC). In comparison to CTRL, the areas of curves (AUCs) of ppr plasma Lys differed: CTRL < CTRL + SBM (p < 0.01) < CTRL + synAA (p < 0.05). For 71 feeds, the estimated pcdCP was correlated with the CP content (p < 0.001), NDSCP (p < 0.001), and ash-free neutral detergent fiber (p < 0.001). A mean neutral detergent insoluble CP content of at least 3-5% can be assumed in horse feed. It is speculated that the predicted availability of Lys from LUC seems to be underestimated by the new protein evaluating system. The influence of chewing and microbiota in vivo needs to be considered in horses.

Investigations on the Effects of Different Calcium Supply Exceeding the Requirements on Mineral Serum Concentrations and Bone Metabolism in Young Warmblood Stallions.

Since mineral supplements for horses commonly contain macro minerals, although the requirement for such is usually covered by roughage-based diets, the aim of this study was to investigate the effects of different dietary calcium levels on mineral serum concentrations and bone metabolism. The trial was conducted in 30 young warmblood stallions (2-3 years) that were divided into two groups for a five-month feeding trial. The groups were fed a hay- and oat-based diet and were either supplied with high (Ca-High) or moderate (Ca-Moderate) calcium excess. While in Ca-High calcium supply was about 2-2.5-fold of the requirement, in Ca-Moderate calcium requirements were slightly surpassed (1.5-1.6-fold). In order to monitor the effects of the different calcium supply, blood samples were taken during the trial and analysed for levels of macro and trace elements as well as concentrations of two bone markers. In Ca-Moderate a trend towards higher phosphorus serum levels compared to Ca-High was observed which was significant at the end of the trial (p = 0.0002). Furthermore, results showed no influence of the diet on bone markers. Results support the idea that forage-based rations for horses do not necessarily have to be supplemented with macro minerals but with trace elements.

Estimation of intestinal permeability in healthy dogs using the contrast medium iohexol.

BACKGROUND: Iohexol is a nonradioactive marker that has been used successfully to test intestinal permeability in humans with inflammatory bowel disease. There is evidence in dogs that iohexol shares a similar permeability pathway as (51)chromium-EDTA, the gold standard marker. OBJECTIVE: The objective of this study was to determine an optimal oral iohexol dosage for an intestinal permeability serum test (IPST) and to use the test to estimate intestinal permeability in healthy dogs. METHODS: Eight clinically healthy dogs free of gastrointestinal, liver, and pancreatic disease were used in the study. Dosages of 0.25, 0.5, 1.0, 2.0, and 4.0 mL/kg of Omnipaque-350 (iohexol) were administered to 2 dogs at weekly intervals. Iohexol concentration was determined in serum samples obtained hourly for 6 hours after administration by high-performance liquid chromatography. Using the optimal dosage, iohexol was administered to 8 dogs twice, 6-36 days (mean 10 days) apart, and coefficients of variation (CVs) for iohexol concentration were calculated. RESULTS: A dosage of 2.0 mL/kg was chosen as optimal for the IPST, based on ease of iohexol detection in serum, intestinal contrast, and clinical effects of iohexol. Following administration of this dose to healthy dogs, mean (+/-SD) serum iohexol concentrations were 8.74+/-4.38, 11.89+/-5.67, 12.40+/-5.47, 9.23+/-5.54, 7.61+/-5.13, and 5.27+/-2.67 microg/mL at 1, 2, 3, 4, 5, and 6 hours after iohexol administration, respectively. CVs between the 2 test days were 28-45%. CONCLUSIONS: Using the iohexol dosage established in this study, the iohexol IPST was easy to perform as a marker for intestinal permeability in dogs. Further studies to establish reference intervals and evaluate the diagnostic value of the iohexol IPST in dogs with gastrointestinal disease are warranted.

Evaluation of the influences of exercise, birth date, and osteochondrosis on plasma bone marker concentrations in Hanoverian Warmblood foals.

OBJECTIVE: To determine whether plasma concentrations of bone turnover markers in growing Hanoverian foals are influenced by age, housing conditions, or osteochondrosis. ANIMALS: 165 healthy foals and 119 foals with osteochondrosis. PROCEDURES: Foals were allocated according to birth date and housing management into groups of early-born (born before March 31, 2001; n = 154 foals, 88 of which were healthy and 66 of which had osteochondrosis) and late-born (born after March 31, 2001; 130 foals, 77 of which were healthy and 53 of which had osteochondrosis) foals. Plasma osteocalcin and carboxyterminal propeptide of type I collagen concentrations were analyzed as markers of bone formation, and carboxyterminal telopeptide of type I collagen concentration was analyzed as a marker of bone resorption. Foals underwent radiographic evaluation to screen for osteochondrosis. RESULTS: Plasma concentrations of osteocalcin, carboxyterminal propeptide of type I collagen, and carboxyterminal telopeptide of type I collagen decreased with age, but these changes were more distinct in late-born foals than in early-born foals. Neither sex nor predisposition to develop osteochondrosis affected the pattern of bone marker changes in either group. CONCLUSIONS AND CLINICAL RELEVANCE: An age-related decrease in concentrations of bone markers was seen during the first 200 days of life. Changes in bone marker concentrations were similar for foals with osteochondrosis and healthy foals. The correlation between the decrease in bone marker concentration and date of birth indicates that there are differences in skeletal development between early- and late-born foals.

[The importance of nutrition counselling in veterinary practice]. / Die Rolle der Ernährung in der Kleintierpraxis.

OBJECTIVE: Collection of epidemiological data about the importance of nutrition counselling and current nutritional knowledge of veterinary surgeons. MATERIAL AND METHODS: A total of 707 veterinarians were interviewed based on a standardized questionnaire. The survey was divided into three sections and comprised the topics person and working environment, level of education and current knowledge regarding nutrition, as well as the significance of nutrition in daily practice. RESULTS: The majority of respondents (91.3%) worked in a practice with up to four veterinarians, where most of the participants (89.3%) treated up to 50 patients daily. On a scale from 1 to 5, the veterinarians evaluated their knowledge with a mean of 2.59 (± 0.798) and the importance of nutrition with a mean of 2.02 (± 0.818). Nutrition consulting was provided for at least half of the patients by 37.3% of the participants. The counselling was performed in different ways. The main reasons for a lack of consultancy despite a current indication were time and labor (37.1%) as well as the low willingness of customers to pay (34.6%). The majority of veterinarians were interested in further training concerning nutrition counselling. CONCLUSION: Nutrition consulting is an important part of daily veterinary practice but there is a demand for further training. CLINICAL RELEVANCE: Although the importance of nutrition is clear, only some of the veterinary surgeons perform consulting on a regular basis. In particular, in case of an existing indication, a nutrition consulting is advisable to ensure maximum success of treatment.

[Effects of a supplementation on sodium chloride or ammonium chloride on urolithic potential in the rabbit]. / Effekte einer Natriumchlorid- oder Ammoniumchloridsupplementierung auf das Harnsteinbildungspotenzial beim Kaninchen.

AIM: Reduction of urolithic potential by means of increased water intake and urine dilution through supplementation of sodium chloride (NaCl) or decrease of urine pH by supplementation of ammonium chloride (NH4Cl) in rabbits. MATERIALS AND METHODS: Sixteen female, 6-month-old dwarf rabbits received the following three feeding regimens in a random order: complete feed without supplements = control; complete feed + 10 g NaCl/kg feed = NaCl; complete feed + 2.5 g NH4Cl/kg feed = NH4Cl. The diets were fed ad libitum over a period of 27 days without roughage. Water was provided ad libitum by a drinker. A 14-day wash-out-period (hay feeding) was performed between the different diets. Blood, faeces, and urine were collected at the beginning of each feeding period, after 21-day adaptation to the respective diet, and after the 3-day collection period. The following parameters were analysed: water and food intake as well as acid-base balance and mineral content in blood, urine, and faeces. RESULTS: NaCl supplementation numerically increased the daily water intake from 40.5 ± 14.4 ml/kg body weight (BW) (control) up to 49.5 ± 14.3 ml/kg BW and significantly increased the daily urine volume from 16.9 ± 7.8 ml/kg BW (control group) to 21.1 ± 7.4 ml/kg BW. The specific gravity of urine samples from NaCl supplementation decreased from 1.060 ± 0.008 to 1.044 ± 0.008. NH4Cl supplementation did not induce significant changes in urine pH, blood acid-base parameters, or calcium retention. Relative supersaturations (RSS) for calcium oxalate and calcium phosphate showed no significant changes after treatment. RSS for struvite increased from 360 ± 735 (after hay feeding) to 3312 ± 6188 on control feeding, 2910 ± 4913 with NaCl supplementation, and 3022 ± 6635 with NH4Cl supplementation (p < 0.05). CONCLUSIONS: NaCl supplementation significantly increased the urine volume and decreased its specific gravity. Therefore, NaCl supplementation might be an additional dietary treatment to increase the elimination of urine crystals in rabbits. NH4Cl supplementation did not induce acidification of the urine.

Impact of diet on 24-hour intragastric pH profile in healthy horses.

An electrode incorporated into a polyethylene hose was introduced under endoscopic control into the stomach of six fasting adult horses for long-lasting pH measurements. The intragastric pH was recorded every four seconds for a period of 24 hours. The Warmblood horses were assigned randomly to receive hay ad libitum (H group); 1.5 kg hay/100 kg BW/day and 1 kg concentrate/100 kg BW/ day (C group) or protocol C plus 75 g pectin-lecithin supplement/100 kg BW/day (P group). The horses were adapted to each diet for 14 days. The 24-hour median pH value for protocol H (2.69) was significantly lower compared to protocol C (3.35) and P (3.44) (p < 0.05). The horses in protocol P had a significant higher percentage (40.1 %) of 24-hour intragastric pH values ≥ 4 than in protocol C (36.2 %) or in protocol H (25.3 %) (p < 0.05).

Effects of deoxynivalenol in naturally contaminated wheat on feed intake and health status of horses.

The present study examined the short-term effects of deoxynivalenol (DON), administered at two different concentrations via a feed preparation using naturally contaminated wheat, on feed intake, liver and kidney metabolism and immunomodulatory properties in horses. Twelve geldings were randomly assigned to one of three dietary treatments for 21 days. DON was provided via naturally contaminated wheat (14.6 ± 6.5 mg DON/kg dry matter). The daily feed intake was adjusted to 4 kg of wheat and 1.7 kg of silage per 100 kg of body weight (BW). Horses were fed one of the following diets: control wheat with 0% contaminated wheat (CON), wheat mixture containing 53 ± 2% of DON-contaminated wheat [low DON intake (LDI)] or wheat mixture containing 78 ± 4% of DON-contaminated wheat [high DON intake (HDI)]. CON, LDI and HDI corresponded to a targeted daily DON intake via the complete ration of <5, 50 and 75 µg/kg BW, respectively. None of the horses demonstrated any clinical signs commonly associated with the intake of DON such as colic or depression. HDI was associated with lower daily wheat intake on day 21. Serum DON concentrations increased with higher DON intake. The non-toxic DON metabolite, deepoxy-deoxynivalenol (DOM-1) was only detected on day 21 of the DON feeding period. No changes in haematological and serum parameters or serum globulins or in the ex vivo proliferation response of peripheral blood mononuclear cells were observed. These results suggest that horses are less sensitive to DON exposure than other domestic species, for example, swine. Therefore, the European Commission guidance value for critical DON concentrations in swine feed (complete diet) of 0.9 mg/kg could be safely applied for rations intended for feeding adult horses as well.

Effects of selenium supplementation on selenium status of farmed fallow deer in outdoor pens.

The study investigated the effects of selenium (Se) supplementation on Se status in farmed fallow deer. Fallow deer were housed on grass pasture and adapted to consume ∼200 g of pelleted grain daily. Animals were divided into two groups. One group received pelleted grain enriched with sodium selenate for 12 weeks (Se+ group, N = 10). Se intake for the first 7 weeks was 0.18 mg/kg dry matter (DM) and 0.32 mg/kg DM for the subsequent 5 weeks. The control group was fed pelleted grain without extra Se (Se− group, N = 9, 0.06-0.08 mg/kg DM). Blood samples were collected at the beginning and the end of the experiment. After the animals were slaughtered, tissue samples were collected for analysis of Se concentrations and Se-dependent glutathione peroxidase 1 (GPx1) activity. In addition, Se-independent α-glutathione-S-transferase (α-GST) activity was analyzed in liver tissue. Se supplementation significantly increased Se levels in plasma and in tissues as follows: liver > spleen > skeletal muscle > myocardium > kidney. Se supplementation also significantly increased GPx1 activity in tissues in the following order: liver > skeletal muscle > spleen = myocardium > kidneys. However, hepatic α-GST activity did not differ between Se+ and Se− groups. As expected, Se supplementation increased blood and tissue Se concentrations and GPx1 activity, which suggests a better antioxidant status. However, the activity of α-GST, an important Se-independent antioxidant enzyme, was not altered, presumably because GPx provided an adequate antioxidant capacity even though Se intake was low.

Ultrasonographic imaging of abomasal milk clotting and abomasal diameter in healthy and diarrheic calves.

In case of diarrhea calves are treated with oral rehydration solutions (ORS), which are known to increase abomasal pH and inhibit milk clotting in vitro. Nevertheless, recent studies have shown that ORS with HCO3(-) ≤ 62 mmol/L do not interfere with abomasal milk clotting in healthy calves. However, in diarrheic calves, feeding ORS and milk simultaneously may disturb abomasal curd formation and exacerbate diarrhea due to faster abomasal passage of ingesta. Therefore, the aim of the present study was to ultrasonographically examine abomasal milk clotting and diameter after feeding milk and milk replacer (MR) with and without ORS to healthy and diarrheic calves. Abomasal curd formation and diameter in healthy and diarrheic calves were ultrasonographically imaged before and after feeding milk, MR and ORS prepared in milk or MR. Feeding mixtures of milk or MR with ORS did not cause any remarkable differences in the ultrasonographic images of abomasal content. Moreover, abomasal milk clotting was not disturbed due to diarrhea. Statistically significant differences of abomasal diameter after feeding between healthy and diarrheic calves indicated that abomasal emptying is delayed in diarrheic calves. Hence, further studies are needed to determine reasons for decelerated abomasal passage in calves suffering from diarrhea.

Long-term effects of intermittent equine parathyroid hormone fragment (ePTH-1-37) administration on bone metabolism in healthy horses.

Intermittent administration of parathyroid hormone (PTH) is an anabolic therapy for osteoporotic conditions in humans. This study evaluated the effects of equine PTH fragment (ePTH-1-37) administration on bone metabolism in 12 healthy horses. Six horses each were treated once daily for 120days with subcutaneous injections of 0.5µg/kg ePTH-1-37 or placebo. Blood was collected to determine ionized calcium (Ca(++)), total Ca (Ca(T)), inorganic phosphorus, serum equine osteocalcin (eOC), carboxy-terminal telopeptide of type I collagen (ICTP), bone-specific alkaline phosphatase, and carboxy-terminal cross-linked telopeptide of type I collagen. Bone mineral density (BMD) was determined with dual X-ray absorptiometry of the metacarpus and calcaneus. Significantly higher blood Ca(++) and plasma Ca(T) concentrations were measured 5h after ePTH-1-37 administration compared to placebo. Higher serum eOC concentrations were found for ePTH-1-37 treatment at days 90 (P<0.05) and 120 (P=0.05). Significantly higher serum ICTP levels were observed with ePTH-1-37 treatment at days 60 and 90. For both study groups, BMD increased significantly in the calcaneus. Long-term use of ePTH-1-37 seemed to have no negative effects on bone metabolism in healthy horses. The absence of undesirable side effects is the premise to ensure safety for further clinical investigations in horses with increased bone resorption processes.

Short-term effects of a moderate fish oil or soybean oil supplementation on postprandial glucose and insulin responses in healthy horses.

The aim of this study was to investigate the effects of fat supplementation on postprandial glycaemic and insulinaemic responses in horses fed a corn meal. Four horses were fed three different diets: (1) cracked corn (CC), (2) cracked corn with soybean oil (CC+SBO), and (3) cracked corn with fish oil (CC+FO). Each diet was adjusted so there was a starch intake of 2 g/kg bodyweight (BW) and an intake of 0.2 mL/kg BW of FO and SBO. The increases in mean plasma glucose and insulin concentrations, peak values, and areas under the curve were similar for all diets (time P<0.05, diet not significant). The addition of FO or SBO at 0.2 mL/kg BW to a starchy meal did not affect acute glucose and insulin responses. To avoid high postprandial glycaemic and insulinaemic responses feeding strategies should be designed primarily to reduce starch intake rather than the addition of fat.

Effect of feeding increasing quantities of starch on glycaemic and insulinaemic responses in healthy horses.

The aim of this study was to investigate the effect of increasing the intake of starch on the glycaemic and insulinaemic responses of horses. A cross-over study design was used in which four horses were fed increasing amounts of a compound feed (0.5-3.5 kg) to provide 0.3, 0.6, 0.8, 1.1, 1.4 and 2 starch/kg bodyweight (BW)/meal. The glycaemic response increased with starch intake (P<0.05), while feeding <1.1 g starch/kg BW resulted in a lowered response, compared to when 1.1-2 g starch/ kg BW was fed (P<0.01). The results suggested that insulin responses may be more appropriate to define the effect of feeding different starch levels than glycaemic responses. A starch intake of <1.1g/kg BW/meal produced only moderate glucose and insulin responses, even though highly processed cereals were used. It is therefore recommended that a starch intake of <1.1 g/kg BW/meal or a meal size of 0.3 kg/100 kg BW (starch content of 30-40%) is used for horses.