Effects of arginine and ornithine supplementation to a high‐protein diet on selected cellular immune variables in adult cats

Background: Dietary protein and amino acid intake and composition can modulate immune function. Objectives: To evaluate the effects of high‐protein intake and arginine and ornithine supplementation on feline immune cells. Animals: Ten healthy cats. Methods: Experimental study. Cats received a high‐protein basal diet as a single daily meal. A crossover design was applied with treatments being basal diet (w/o); basal diet with arginine supplementation (+50, 75, 100% compared to the arginine provision by the basal diet; Arg 1‐3); and basal diet with ornithine supplementation (+100, 150, 200% compared to the arginine provision by the basal diet; Orn 1‐3). Blood samples were collected at the end of each 11‐day treatment period. Results: Mitogen‐stimulated proliferative activity of blood leukocytes revealed a quadratic effect for the dietary supplementation of arginine (P = .02) and ornithine (P = .03) (means for ConA‐stimulation: w/o = 6.96; Arg 1 = 9.31; Arg 2 = 11.4; Arg 3 = 8.04; Orn 1 = 15.4; Orn 2 = 9.43; Orn 3 = 9.28; pooled SEM: 0.96). The number (% gated) of phagocytic granulocytes linearly decreased with increasing dietary concentrations of arginine (P = .05) and ornithine (P = .03) (means: w/o = 95.5; Arg 1 = 93.0; Arg 2 = 92.5; Arg 3 = 92.6; Orn 1 = 92.6; Orn 2 = 92.6; Orn 3 = 91.5; pooled SEM = 0.44). Conclusions and Clinical Importance: This study could demonstrate immunomodulating properties of dietary arginine and ornithine in cats

Urinary calcium and oxalate excretion in healthy adult cats are not affected by increasing dietary levels of bone meal in a canned diet

This study aimed to investigate the impact of dietary calcium (Ca) and phosphorus (P), derived from bone meal, on the feline urine composition and the urinary pH, allowing a risk assessment for the formation of calcium oxalate (CaOx) uroliths in cats. Eight healthy adult cats received 3 canned diets, containing 12.2 (A), 18.5 (B) and 27.0 g Ca/kg dry matter (C) and 16.1 (A), 17.6 (B) and 21.1 g P/kg dry matter (C). Each diet was fed over 17 days. After a 7 dayś adaptation period, urine and faeces were collected over 2×4 days (with a two-day rest between), and blood samples were taken. Urinary and faecal minerals, urinary oxalate (Ox), the urinary pH and the concentrations of serum Ca, phosphate and parathyroid hormone (PTH) were analyzed. Moreover, the urine was microscopically examined for CaOx uroliths. The results demonstrated that increasing levels of dietary Ca led to decreased serum PTH and Ca and increased faecal Ca and P concentrations, but did not affect the urinary Ca or Ox concentrations or the urinary fasting pH. The urinary postprandial pH slightly increased when the diet C was compared to the diet B. No CaOx crystals were detected in the urine of the cats. In conclusion, urinary Ca excretion in cats seems to be widely independent of the dietary Ca levels when Ca is added as bone meal to a typical canned diet, implicating that raw materials with higher contents of bones are of subordinate importance as risk factors for the formation of urinary CaOx crystals

Effects of Dietary Arginine, Ornithine, and Zeolite Supplementation on Uremic Toxins in Cats

To test if arginine and ornithine, both components of the Krebs-Henseleit cycle, or zeolite, a potential ammonium absorber, can modulate the excretion of harmful bacterial metabolites, intestinal microbial protein fermentation was stimulated by feeding a high-protein (60.3%) diet as a single daily meal to 10 adult cats. The diet was supplemented without or with arginine (+50, 75, 100% compared to arginine in the basal diet), ornithine (+100, 150, 200% compared to arginine in the basal diet), or zeolite (0.125, 0.25, 0.375 g/kg body weight/day). The cats received each diet for 11 days. Urine, feces, and blood were collected during the last 4 days. Arginine and ornithine enhanced the postprandial increase of blood urea, but renal urea excretion was not increased. Zeolite decreased renal ammonium excretion and fecal biogenic amines. The data indicate an increased detoxification rate of ammonia by arginine and ornithine supplementation. However, as urea was not increasingly excreted, detrimental effects on renal function cannot be excluded. Zeolite had beneficial effects on the intestinal nitrogen metabolism, which should be further evaluated in diseased cats. Clinical studies should investigate whether dietary arginine and ornithine might improve hepatic ammonia detoxification or could be detrimental for renal function

The use of Moringa oleifera in ruminant feeding and its contribution to climate change mitigation

Livestock production in developing countries faces several difficulties such as a general shortage of feed resources, regional availability, and quality. Climate change further exacerbates these problems, leading to a massive reduction in ruminant productivity. Therefore, there is a need for the use of adaptable and resilient forage plants that can also contribute to reducing greenhouse gases. The Moringa oleifera tree is well known as an agroforestry tree and has adapted to growing in harsh conditions. It produces a high amount of biomass in a short period and contains high levels of nutrients and biologically active components. All parts of the Moringa tree are valuable and have multiple benefits and applications. Therefore, Moringa oleifera has great potential and can be used as a forage crop, storing carbon dioxide (CO2) and improving ruminant performance and the livelihoods of farmers in the tropics. This article aimed to present the results and findings of studies related to the use of Moringa in ruminant feed (cattle, sheep, and goats) and its contribution to climate protection. Several studies highlighted that M. oleifera can be used as green fodder either individually or in combination with other crops or concentrate feeds to improve the performance of ruminants, such as the growth rate, milk yield, and milk constituents, without negatively impacting animal health. This improvement in performance could be attributed to the favorable nutrient content in M. oleifera, delivering proteins in conjunction with bioactive compounds such as alkaloids, flavonoids, phenolics, glucosinolates, carotenoids, sterols, saponins, phenolic acids, tannins, and isothiocyanates. Furthermore, it has been shown that this plant can be produced in high yields and thus might be an excellent carbon dioxide sink to absorb and utilize carbon dioxide, reducing the anthropogenic load of carbon dioxide in the atmosphere. In addition, feeding cattle and other ruminants with M. oleifera leaves or seeds significantly decreases ruminal methane emissions, which could contribute to adapting to climate-friendly farming. Thus, the use of Moringa can make a sustainable contribution to strengthening animal production, especially in countries with limited feed resources

Optimization of Production Parameters for Probiotic Lactobacillus Strains as Feed Additive

In animal nutrition, probiotics are considered as desirable alternatives to antibiotic growth promoters. The beneficial effects of probiotics primarily depend on their viability in feed, which demands technical optimization of biomass production, since processing and storage capacities are often strain-specific. In this study, we optimized the production parameters for two broiler-derived probiotic lactobacilli (L. salivarius and L. agilis). Carbohydrate utilization of both strains was determined and preferred substrates that boosted biomass production in lab-scale fermentations were selected. The strains showed good aerobic tolerance, which resulted in easier scale-up production. For the freeze-drying process, the response surface methodology was applied to optimize the composition of cryoprotective media. A quadratic polynomial model was built to study three protective factors (skim milk, sucrose, and trehalose) and to predict the optimal working conditions for maximum viability. The optimal combination of protectants was 0.14g/mL skim milk/ 0.08 g/mL sucrose/ 0.09 g/mL trehalose (L. salivarius) and 0.15g/mL skim milk/ 0.08 g/mL sucrose/ 0.07 g/mL (L. agilis), respectively. Furthermore, the in-feed stabilities of the probiotic strains were evaluated under different conditions. Our results indicate that the chosen protectants exerted an extensive protection on strains during the storage. Although only storage of the strains at 4 °C retained the maximum stability of both Lactobacillus strains, the employed protectant matrix showed promising results at room temperature

Dietary inulin affects the intestinal microbiota in sows and their suckling piglets

Background: Several studies have focused on the effects of dietary inulin on the intestinal microbiota of weaned piglets. In the present study, inulin was added to a diet for gestating and lactating sows, expecting not only effects on the faecal microbiota of sows, but also on the bacterial cell numbers in the gastrointestinal tract of their piglets during the suckling period. Sows were fed a diet without (n = 11) or with (n = 10) 3% inulin, and selected bacterial groups were determined in their faeces ante and post partum. Suckling piglets, 8 per group, were euthanised on day 10 after birth to analyse digesta samples of the gastrointestinal tract. Results: Dietary inulin increased the cell numbers of enterococci, both, in the faeces of the sows during gestation and lactation, and in the caecum of the piglets (P ≤ 0.05). Moreover, higher cell numbers of eubacteria (stomach) and C. leptum (caecum), but lower cell numbers of enterobacteria and L. amylovorus (stomach) were detected in the digesta of the piglets in the inulin group (P ≤ 0.05). Conclusions: In conclusion, inulin seems to have the potential to influence the gastrointestinal microbiota of suckling piglets through the diet of their mother, showing the importance of the mother-piglet couple for the microbial development. Early modulation of the intestinal microbiota could be especially interesting with regard to the critical weaning time

Influence of high- and low-fermentable dietary fibres in sows' diet on the colostrum potential against Clostridioides difficile toxin-induced effects in IPEC-J2 cells

Sow colostrum has been reported to protect the IPEC-J2 cells and piglet colon tissues from detrimental effect of Clostridioides difficile toxins. Since dietary fibre can influence the colostrum composition in sows, we hypothesised that it can also differentially affect the colostrum potential against C. difficile toxin-induced effects in IPEC-J2. IPEC-J2 were incubated with colostrum from sows fed either high-fermentable sugar beet pulp (SBP) or low-fermentable lignocellulose (LNC) fibres and in combination with the toxins and analysed by trans-epithelial electrical resistance (TEER) and cell viability using propidium iodide in flow cytometry. Toxins drastically decreased the integrity of IPEC-J2. Colostrum from the sows fed either SBP or LNC exerted protective effect against toxins on IPEC-J2 integrity and this effect was numerically superior in the SBP group. Differences in the percentages of TEER between different treatments were noted after 2 h (p = 0.043), 3 h (p = 0.017) and 4 h (p = 0.017) of incubation and a tendency for differences was noted after 5 h of incubation (p = 0.071). Colostrum from either SBP- or LNC-fed sows did not protect the IPEC-J2 from toxin-induced death. Colostrum of the sows fed either high-fermentable or low-fermentable fibres has a potential to protect IPEC-J2 from the loss of integrity, which may be important in protection from C. difficile-infection development in neonatal piglets

The Impact of Dietary Zinc Oxide on the Bacterial Diversity of the Small Intestinal Microbiota of Weaned Piglets

Dietary zinc oxide is often used in pharmacological concentrations to promote health as well as performance of weaned piglets due to its bacteriostatic effects. This study was conducted to provide an in depth analysis of the bacterial composition in weaned piglets fed different amounts of dietary zinc oxide. Piglets were fed diets containing 57 (low), 164 (medium) or 2425 (high) mg/kg dietary zinc. Zinc above the basal dietary level was supplied from analytical grade zinc oxide (ZnO). DNA was extracted from stomach and ileum digesta samples of 32 and 53d old animals (n=4 per group) and used to generate bar-coded 16S ribosomal DNA amplicons for deep sequencing analysis. A total of 9 phyla, 40 orders, 75 families and 328 genera were detected in 8.76 x 105 sequencing reads. Firmicutes, Bacteroidetes and Proteobacteria were the dominant phyla, but no significant differences between treatment groups were observed. Lactobacillales (16.3-59.9%), Bacteroidales (2.2-59.1%), Clostridiales (0.05-70.2%) and Selenomonadales (2.6-17.5%) were found as the dominating order. Noteworthy changes on the order level were found for numerically or significantly increased ratios of Clostridiales, but significantly decreased Lactobacillales in the high dietary zinc group. The bacterial diversity for the high dietary zinc diet was significantly higher for the total microbiota than the medium or low zinc diet. However, Lactobacillales diversity decreased, while Clostridiales and Enterobacteriales diversity increased significantly. Principal component analysis confirmed changes in the microbiota, most notably for the high dietary zinc treatment. This study has shown that pharmacological doses of high dietary zinc can drastically alter the bacterial composition and development of the microbiota in weaned piglets. The quantitative shift of bacterial groups due to high dietary zinc was most pronounced one week after weaning, while the more developed microbiota in older animals seemed to be able to adapt to high concentrations of dietary zinc

Impact of dietary supplementation of l-Arginine, l-Glutamine, and the combination of both on nursing performance of multiparous sows

Dietary supplementation with arginine (Arg) or glutamine (Gln) has been considered as an option to improve nursing performance in reproductive sows. This study investigated whether a low-level supplementation of Arg or Gln or a blend of both could modify milk nutrients and improve piglets’ growth beyond weaning. Seventy-two multiparous sows were assigned to four groups: one group fed a control diet, three treatment groups fed the control diet supplemented with either 0.35% Arg, 0.35% Gln, or both, from day 108 of gestation until weaning at day 26 of lactation. Immediately after birth, the litters were cross fostered to 13 piglets and monitored until 2 wk after weaning. Sows body condition and litter growth were assessed. Colostrum and milk samples were collected for nutrient analyses. Plasma concentrations of insulin-like growth factor 1 (IGF-1) around weaning were determined in sows and two representative piglets per litter. Supplementing Gln or the combination of Arg and Gln had no effect on the parameters studied. Arg supplementation increased weaning weight, while decreasing the variation of piglet weights 2 wk after weaning. There was no correlation with plasma IGF-1 since the hormone was not altered in sows or piglets. The colostral concentration of fat tended to increase in the Arg-group, whereas protein, lactose, energy, and polyamine concentrations remained unaffected. Milk samples obtained on day 12 and 25 of lactation were not influenced by dietary treatment. The data indicate that there might be a window of opportunity, explicitly at the onset of lactation, for dietary intervention by maternal dietary Arg supplementation

In vitro conjugation kinetics of AmpC, broad spectrum and extended-spectrum beta-lactamase-producing Escherichia coli donors and various Enterobacteriaceae recipients

Background: Extended spectrum beta-lactamase (ESBL)-producing enterobacteria pose a major hazard to public health. Due to the possibility of genetic transfer, ESBL genes might spread to pathogenic enterobacterial strains. Thus, information on possible genetic transfer between enterobacteria is of high interest. It was therefore the aim of this in vitro study to screen the capacity of a wide range of Enterobacteriaceae for differences in conjugation at different time points with five ESBL-producing Escherichia coli strains. Results: Conjugation frequencies for five potential E. coli donor strains producing the enzymes CTX-M-1, CTX-M-15, SHV-12, TEM-1, TEM-52 and CMY-2, and six potential recipient strains commonly detected in the gastrointestinal tract of poultry (E. coli, Serratia marcescens subsp. marcescens, Enterobacter cloacae, Salmonella (S.) enterica serovar Typhimurium and Proteus mirabilis) were obtained. Different combinations of donor and recipient strains were co-incubated for between 0 and 22 h and spread on selective agar. Conjugation frequencies were calculated as transconjugants per donor. Some donor and recipient strain combinations did not perform plasmid transfer within 22 h. Hence, the recipient Proteus mirabilis did not accept plasmids from any of the given donors and the E. coli ESBL10716 donor was unable to transfer its plasmid to any recipient. Enterobacter cloacae only accepted the plasmids from the donors E. coli ESBL10708 and E. coli ESBL10716 while E. coli ESBL10708 did not transfer its plasmid to Serratia marcescens subsp. marcescens. E. coli IMT11716 on the other hand did not perform conjugation with the donor E. coli ESBL10689. The remaining mating pairs differed in conjugation frequency, ranging from 10− 5 to 10− 9 transconjugants/donor. The earliest conjugation events were detected after 4 h. However, some mating pairs turned positive only after 22 h of coincubation. Conclusion: A suitable mating pair for future in vivo studies to combat transfer of antibiotic resistance to pathogenic bacteria in broiler chicken was determined. The results of this study also suggest that the kinetic of conjugation differs between mating pairs and is independent of species origin. This should be considered when performing conjugation experiments