Effects of Omega-3 Polyunsaturated Fatty Acids on the Formation of Adipokines, Cytokines, and Oxylipins in Retroperitoneal Adi-Pose Tissue of Mice.

Oxylipins and specialized pro-resolving lipid mediators (SPMs) derived from polyunsaturated fatty acids (PUFAs) are mediators that coordinate an active process of inflammation resolution. While these mediators have potential as circulating biomarkers for several disease states with inflammatory components, the source of plasma oxylipins/SPMs remains a matter of debate but may involve white adipose tissue (WAT). Here, we aimed to investigate to what extent high or low omega (n)-3 PUFA enrichment affects the production of cytokines and adipokines (RT-PCR), as well as oxylipins/SPMs (liquid chromatography-tandem mass spectrometry) in the WAT of mice during lipopolysaccharide (LPS)-induced systemic inflammation (intraperitoneal injection, 2.5 mg/kg, 24 h). For this purpose, n-3 PUFA genetically enriched mice (FAT-1), which endogenously synthesize n-3 PUFAs, were compared to wild-type mice (WT) and combined with n-3 PUFA-sufficient or deficient diets. LPS-induced systemic inflammation resulted in the decreased expression of most adipokines and interleukin-6 in WAT, whereas the n-3-sufficient diet increased them compared to the deficient diet. The n-6 PUFA arachidonic acid was decreased in WAT of FAT-1 mice, while n-3 derived PUFAs (eicosapentaenoic acid, docosahexaenoic acid) and their metabolites (oxylipins/SPMs) were increased in WAT by genetic and nutritional n-3 enrichment. Several oxylipins/SPMs were increased by LPS treatment in WAT compared to PBS-treated controls in genetically n-3 enriched FAT-1 mice. Overall, we show that WAT may significantly contribute to circulating oxylipin production. Moreover, n-3-sufficient or n-3-deficient diets alter adipokine production. The precise interplay between cytokines, adipokines, and oxylipins remains to be further investigated.

Effects of dietary zinc chloride hydroxide and zinc methionine on the immune system and blood profile of healthy adult horses and ponies.

The effects of dietary zinc on the immune function of equines have not been evaluated in detail so far. In the present study, eight healthy adult ponies and two healthy adult horses were fed a diet supplemented with either zinc chloride hydroxide or zinc methionine in six feeding periods of four weeks each (according to maintenance zinc requirement, 120 mg zinc/kg dry matter, and 240 mg zinc/kg dry matter, for both dietary zinc supplements, respectively). All animals received the six diets, with increasing amounts of zinc chloride hydroxide in the feeding periods 1-3, and with increasing amounts of zinc methionine in the feeding periods 4-6. At the end of each feeding period, blood samples were collected for a blood profile and the measurement of selected immune variables. Increasing dietary zinc chloride hydroxide doses increased the glutathione concentrations in the erythrocyte concentrate and the glutathione peroxidase activity in the erythrocyte lysate, decreased the numbers of total leukocytes and granulocytes in the blood, and also decreased the interleukin-2 concentrations in the plasma of the animals. The dietary supplementation of increasing doses of zinc methionine enhanced the mitogen-stimulated proliferative activity of peripheral blood mononuclear cells, and decreased the glutathione concentrations in the erythrocyte concentrate and the glutathione peroxidase activity in the plasma of the animals. The percentage of blood monocytes with oxidative burst after in vitro stimulation with E. coli decreased with increasing dietary zinc concentrations, independently of the zinc compound used. The blood profile demonstrated effects of the zinc supplements on the red blood cells and the bilirubin metabolism of the horses and ponies, which require further investigation. Overall, high doses of dietary zinc modulate the equine immune system, for the most part also depending on the zinc compound used.

Effects of dietary cellobiose on the intestinal microbiota and excretion of nitrogen metabolites in healthy adult dogs.

In order to evaluate the potential prebiotic effects of cellobiose, 10 healthy adult research beagle dogs received a complete diet containing 0, 0.5 and 1 g cellobiose/kg bodyweight (BW)/day. At the end of each feeding period, faeces, urine and blood of the dogs were collected. The results demonstrated a significant increase of faecal lactate concentrations, indicating a bacterial fermentation of cellobiose in the canine intestine. Along with this, a dose-dependent linear increase of the relative abundance of Lactobacillaceae in the faeces of the dogs was observed (p = 0.014). In addition, a dose-dependent increase (p < 0.05) of Alloprevotella, Bacteroides and Prevotella, and a linear decrease for unidentified Lachnospiraceae (p = 0.011) was observed when cellobiose was added to the diet, although the relative abundance of these genera was low (<1%) among all groups. The faecal pH was not affected by dietary cellobiose. Cellobiose seemed to modulate the excretion of nitrogen metabolites, as lower concentrations of phenol (p = 0.034) and 4-ethylphenol (p = 0.002) in the plasma of the dogs were measured during the supplementation periods. Urinary phenols and indoles, however, were not affected by the dietary supplementation of cellobiose. In conclusion, cellobiose seems to be fermented by the intestinal microbiota of dogs. Although no effect on the faecal pH was detected, the observed increase of microbial lactate production might lower the pH in the large intestine and consecutively modulate the intestinal absorption of nitrogen metabolites. Also, the observed changes of some bacterial genera might have been mediated by increased intestinal lactate concentrations or a higher relative abundance of lactobacilli. Whether these results could be considered as a prebiotic effect and used as a dietetic strategy in diseased animals to improve gut function or hepatic and renal nitrogen metabolism should be evaluated in future studies.

Impact of the dietary inclusion of dried food residues on the apparent nutrient digestibility and the intestinal microbiota of dogs.

The use of food residues for animal nutrition might imply ecological and economic advantages; however, their effects as a potential ingredient have not yet been evaluated in dogs. In the present study, four diets with 0, 5, 10 and 15% dried food residues (DFR), derived from hotel catering, were fed to 10 healthy adult dogs. At the end of each three-week feeding period, faeces and blood were collected. The apparent nutrient digestibility was calculated by the dietary inclusion of titanium dioxide as an inert marker. The results demonstrated that the apparent crude protein digestibility and ether extract digestibility decreased with increasing amounts of DFR in the diets (p < 0.05). In addition, an increase of the faecal concentrations of acetic acid, propionic acid, n-butyric acid and total short-chain fatty acids (SCFA) was observed (p < 0.05). Faecal ammonium and lactate concentrations, as well as plasma phenol and indole concentrations, were not linearly affected by the dietary inclusion of DFR. The relative abundance of Fusobacteria in the faeces of the dogs decreased, and the relative abundance of Actinobacteria and Bacteroidetes increased with increasing amounts of DFR in the diets (p < 0.05). In conclusion, the DFR seemed to be intensively fermented by the intestinal microbiota of the dogs, as indicated by the increased faecal SCFA concentrations and the shifts in the composition of the faecal microbiota. Dietary inclusion levels of up to 5% can be recommended based on our results, as the observed lower apparent crude protein and ether extract digestibility might limit the use of food residues for dogs at higher amounts.

Influence of protein concentration and quality in a canned diet on urine composition, apparent nutrient digestibility and energy supply in adult cats.

BACKGROUND: Protein concentration and quality in cat food can vary considerably, and the impact on feline urine composition and nutrient supply is of high practical relevance. In the present study, 6 canned diets with varying protein concentrations and qualities were fed to 10 healthy adult cats. Protein quality in the diet differed depending on the amount of collagen-rich ingredients. Hydroxyproline concentrations were 2.56-4.45 g/kg dry matter in the high quality and 3.76-9.44 g/kg dry matter in the low quality diets. Protein levels were 36.2, 43.3 and 54.9% in the high quality and 36.7, 45.0 and 56.1% in the low quality groups. Each diet was fed for 6 weeks, using a randomized cross-over design. In the last 2 weeks of each feeding period, urine and faeces of the cats were collected. RESULTS: Renal calcium (Ca), oxalate (Ox) and citrate excretion were unaffected by the dietary protein concentration, possibly mediated by a high urine volume (24.2-34.2 ml/kg bodyweight (BW)/day) in all groups. However, renal Ox excretion was lower when the high quality diets were fed (P = 0.013). Urinary relative supersaturation (RSS) with calcium oxalate (CaOx) was low in general, but reduced in the high quality groups (P = 0.031). Urinary RSS values for magnesium ammonium phosphate (MAP) were high (2.64-5.00) among all groups. Apparent digestibility of crude protein and most minerals was unaffected by the different diets. Feed intake was higher in the low quality groups (P = 0.026), but BW of the cats did not differ depending on dietary protein quality. BW of the cats increased with increasing dietary protein concentrations (P = 0.003). CONCLUSION: In conclusion, a high protein canned diet might not be a specific risk factor for CaOx urolith formation in cats. In contrast, all diets resulted in high RSS MAP values, which might be critical concerning MAP crystallization. Protein quality had a minor, but significant impact on urine composition, necessitating further research on this subject. A lower energy supply when feeding a low protein quality can be assumed. Changes in BW were only small and require a careful interpretation.

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.

Effects of potassium chloride and potassium bicarbonate in the diet on urinary pH and mineral excretion of adult cats.

Low dietary K levels have been associated with increasing renal Ca excretion in humans, indicating a higher risk of calcium oxalate (CaOx) urolith formation. Therefore, the present study aimed to investigate whether dietary K also affects the urine composition of cats. A total of eight adult cats were fed diets containing 0·31 % native K and 0·50, 0·75 and 1·00 % K from KCl or KHCO3 and were evaluated for the effects of dietary K. High dietary K levels were found to elevate urinary K concentrations (P<0·001). Renal Ca excretion was higher in cats fed the KCl diets than in those fed the KHCO3 diets (P=0·026), while urinary oxalate concentrations were generally lower in cats fed the KCl diets and only dependent on dietary K levels in cats fed the KHCO3 diets (P<0·05). Fasting urine pH increased with higher dietary K levels (P=0·022), reaching values of 6·38 (1·00 % KCl) and 7·65 (1·00 % KHCO3). K retention was markedly negative after feeding the cats with the basal diet (-197 mg/d) and the 0·50 % KCl diet (-131 mg/d), while the cats tended to maintain their balance on being fed the highest-KCl diet (-23·3 mg/d). In contrast, K from KHCO3 was more efficiently retained (P=0·018), with K retention being between -82·5 and 52·5 mg/d. In conclusion, the dietary inclusion of KHCO3 instead of KCl as K source could be beneficial for the prevention of CaOx urolith formation in cats, since there is an association between a lower renal Ca excretion and a generally higher urine pH. The utilisation of K is distinctly influenced by the K salt, which may be especially practically relevant when using diets with low K levels.

Liver and kidney concentrations of strontium, barium, cadmium, copper, zinc, manganese, chromium, antimony, selenium and lead in cats.

BACKGROUND: In order to provide new knowledge on the storage of strontium (Sr), barium (Ba), cadmium (Cd), copper (Cu), zinc (Zn), manganese (Mn), chromium (Cr), antimony (Sb), selenium (Se) and lead (Pb) in the feline organism, we measured the concentrations of these elements in the liver, renal cortex and renal medulla, evaluating also the impact of age, sex or the occurrence of a chronic kidney disease (CKD). The element concentrations in the tissues of 47 cats (22 male; 25 female; aged between 2 months and 18 years) were measured using inductively coupled plasma mass spectrometry. RESULTS: Cu, Zn and Mn were the highest in the liver, followed by the renal cortex and the renal medulla. The Cd concentrations were lower in the renal medulla compared to the renal cortex and the liver, and Sr was higher in the renal medulla compared to the liver. The Se concentrations in the cortex of the kidneys were higher than in the medulla of the kidneys and in the liver. Higher Cd concentrations were measured in the renal cortex of female cats, while no further gender-related differences were observed. Except for Cr, Sb and Se, age-dependencies were detected for the storage of all elements. The occurrence of a CKD also affected the storage of the elements, with lower concentrations of Ba (renal medulla), Zn (renal cortex; renal medulla) and Mn (liver; renal medulla), but higher Cd concentrations (liver; renal cortex) in diseased cats. CONCLUSIONS: In conclusion, the present results provide new information on the accumulation of specific elements in the feline liver and kidneys, demonstrating a dependency on age and an impaired kidney function, but not on the sex of the animals.

Zinc Status of Horses and Ponies: Relevance of Health, Horse Type, Sex, Age, and Test Material.

Little is known about the animal- and diet-related factors that could interfere with the plasma zinc (Zn) concentrations of equines. Additionally, the adequacy of plasma to reflect changes in the Zn intake is unclear. In the first part of this study, the plasma Zn concentrations of hospitalized horses and ponies (n = 538) were measured and evaluated for the impact of the age, sex, horse type, and internal diseases of the animals. In the second part, the effects of increasing dietary Zn chloride hydroxide and Zn methionine supplementations were assessed on the plasma and mane hair Zn concentrations of healthy horses (n = 2) and ponies (n = 8). Part 1: The age, sex, and horse type did not influence the plasma Zn concentrations. No effect of internal diseases was observed, with the exception of higher plasma Zn concentrations in animals with metabolic disorders compared to the control group (p < 0.05). Part 2: Both Zn supplements dose-dependently increased the Zn concentrations in the mane hair (p = 0.003), but not in the plasma of the horses and ponies. In conclusion, the plasma Zn concentrations were widely unaffected by nutritional and non-nutritional factors in equines, while mane hair samples better reflected the dietary Zn supply.

Effects of the Protein Concentration and Quality in a Canned Diet on the Fecal Microbiota of Healthy Adult Cats.

Feline diets can markedly differ in their protein concentration and quality, which might also affect the intestinal microbiota of cats. In the present study, 6 canned diets, differing in their protein quality (high/low, achieved by varying amounts of meat and collagen-rich ingredients) and concentration (high quality/low quality: 36.2/36.7% in dry matter (DM), 43.3/45.0% in DM and 54.9/56.1% in DM), were fed to 10 healthy adult cats for 6 weeks each. At the end of the feeding periods, fecal samples were collected to analyze the microbiota (16S rDNA sequencing) and bacterial metabolites. Increasing dietary protein concentrations increased the relative abundance of Fusobacterium and Bacteroides as well as the concentrations of ammonium and n-valerate in the feces of the cats, independently of the dietary protein quality. A lower dietary protein quality was accompanied by a higher evenness index and a higher relative abundance of Fusobacteria and Bacteroidetes in the feces when compared to the feeding of the high protein quality diets. A promotion of bacterial proteolytic activity and, in particular, increased intestinal ammonium concentrations might be undesired effects of high protein intakes in cats. Whether the long-term feeding of those diets could be critical for feline health requires further investigation.

Impact of Dietary Zinc Chloride Hydroxide and Zinc Methionine on the Faecal Microbiota of Healthy Adult Horses and Ponies.

Zinc supplements are often used in equine nutrition to support skin and hoof quality or the immune function. However, no data on the effects of dietary zinc on the intestinal microbiota of horses and ponies are available so far. In the present study, varying dietary zinc concentrations (maintenance (4 mg/kg BW0.75/day), 120 mg/kg dry matter (DM)/day and 240 mg/kg DM/day) were achieved by the supplementation of either zinc chloride hydroxide or zinc methionine (six treatment periods of 4 weeks each). Eight healthy adult ponies and two healthy adult horses were included, and faecal samples were collected at the end of each treatment period to analyse the microbiota (16S rDNA sequencing) and microbial metabolites. With increasing dietary zinc concentrations, the richness of the faecal microbiota decreased, independently of the zinc compound used. In addition, a decrease of the relative abundance of Bacteroidetes and Fibrobacteres as well as of acetate and total short-chain fatty acids in the faeces of the animals was observed at high zinc intakes. Effects on the bacterial order, family and genus level were also detected, which were partly more pronounced for zinc chloride hydroxide. It can be concluded that high dietary zinc levels decrease the richness and fermentative activity of the faecal microbiota of horses and ponies. Although all animals were healthy throughout the study, the effects could be critical for gut health, and deserve more research. The detected differences between the zinc compounds used indicate differences in the bioavailability of organic and inorganic zinc sources in equines.

In vitro digestion and microbial fermentation of dried food residues, a potential "new" component for pet food, and different non-digestible carbohydrate sources.

Food residues are often fed to dogs in private households and might also be a potential "new" ingredient for pet food in the future. As food residues might contain not only digestible, but also fermentable substrates, an effect on the intestinal microbiota can be assumed. In the present study, two batches of dried food residues (DFR) collected from hotels in Crete were microbially fermented in an in vitro batch culture system with canine fecal inoculum: non-sterile DFR including meat (DFRm), sterile DFR including meat (DFRms) and sterile DFR without meat (DFRwms). Different non-digestible carbohydrate sources (beet pulp, wheat bran, inulin, carrot pomace, brewer´s spent grains, cellulose and lignocellulose) were included for comparison. Inulin, cellulose and lignocellulose were only used as raw materials, while the other test substrates were incubated as raw and enzymatically pre-digested substrates. After incubation for 24 hours, the raw food residues markedly increased the concentrations of bacterial metabolites in the fermenters, although smaller effects were observed for the DFRwms. When the enzymatically pre-digested food residues were incubated, the effects were more pronounced for the DFRms and DFRwms. In general, when compared with the other test substrates, the food residues were microbially fermented to a comparable or partly higher extent. Interestingly, high n-butyrate concentrations were measured in the inocula, both after incubation of the raw and pre-digested food residues. In conclusion, the food residues contained enzymatically digestible and microbially fermentable substrates. If considered as a potential future ingredient for pet food, a standardization of the collection and processing of food residues might be necessary in order to reduce compositional variability and varying effects on the intestinal microbiota.

A Pilot Study on the Urine Proteome of Cats Fed a High-Protein Complete Diet, Supplemented with or without Arginine, Ornithine or Zeolite.

Proteome analyses can be used to detect biomarkers for the healthy and diseased organism. However, data in cats are scarce, and no information is available on the potential impact of nutritional interventions on the feline urine proteome. In the present study, a label-free shotgun proteomics approach was performed to investigate the urinary proteins of four healthy adult cats. Each animal received a high-protein complete diet without (w/o) or with supplements that could affect the protein metabolism: arginine (+100% compared to the arginine concentration in the w/o diet), ornithine (+200% compared to the arginine concentration in the w/o diet) or zeolite (0.375 g/kg body weight/day). Our results demonstrate a huge number of proteins in the urine of cats (516 ± 49, 512 ± 39, 399 ± 149 and 455 ± 134 in the w/o, arginine, ornithine and zeolite group, respectively), which are associated with several biological processes. In addition, up- and downregulated urinary proteins could be detected in the dietary supplementation periods. Overall, the present pilot study provides basic data on the urine proteome of healthy adult cats. With increasing information, the numerousness of urinary proteins implies the potential to identify biomarkers and metabolic pathways in the feline organism.

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.

Impact of Dietary Cellobiose on the Fecal Microbiota of Horses.

Cellobiose is a disaccharide with potential prebiotic effects, as demonstrated in different animal species, but not yet in horses. It was, therefore, the aim of the present study to evaluate the impact of dietary cellobiose on the fecal microbiota of horses. Eight healthy adult horses and two ponies were included in this study. The animals received a diet without or with 10 g and 20 g cellobiose per day for 14 days each. At the end of the feeding periods, fresh fecal samples were collected to measure bacterial metabolites and the microbial composition. For the microbiota analysis, 16S rRNA gene sequencing was used. Cellobiose was well accepted and tolerated by the animals. The lowest fecal concentrations of D-lactate, propionic acid, i-valeric acid, and total short-chain fatty acids were measured at the dose 10 g cellobiose per horse per day (quadratic effect: P < .05). A dose-dependent increase of the relative abundance of Firmicutes (P = .049), Coriobacteriales (P < .001), and Clostridium (P = .031) could be detected. In addition, a dose-dependent decrease of the relative abundance of Bacteroidetes (P = .035) was observed. In conclusion, the increase of Coriobacteriales and Clostridium indicates a bacterial fermentation of cellobiose in the equine intestine, as members of both groups exert saccharolytic activity. As clostridia have previously been assumed to be a key component of the intestinal microbiota in horses, the observed increase of Clostridium in the feces might indicate beneficial and potentially prebiotic effects of cellobiose in horses. However, this finding requires further investigation, particularly with regard to the Clostridium species that have been promoted by dietary cellobiose.

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.

[Acceptance, tolerance and apparent nutrient digestibility of complete diets based on larvae meal of Hermetia illucens in cats]. / Akzeptanz, Verträglichkeit und scheinbare Nährstoffverdaulichkeit von Alleinfuttermitteln auf Basis von Hermetia-illucens-Larvenmehl bei Katzen.

OBJECTIVE: Insect meal can be used as a component of cat food for several reasons. In addition to veterinary indications, particularly for animals suffering from food allergy, economic or ecological reasons might be relevant. Because no data in cats are available on the acceptance, tolerance or nutrient digestibility of a diet based on insect protein, it was the aim of the present study to evaluate these aspects more closely. MATERIAL AND METHODS: Ten healthy, adult cats received two different diets (A, B) based on larvae meal of the black soldier fly (Hermetia illucens) for 6 weeks each. At the end of each feeding period, the faeces of the cats were collected and analysed. Both diets contained titanium dioxide as an indigestible marker to calculate the apparent nutrient digestibilities. RESULTS: The results demonstrated a generally good food tolerance and acceptance. However, one cat displayed emesis after the ingestion of diet A and subsequently refused the diet, thus this animal was removed from the study. With regard to diet B, no emesis of the cats was observed. However, three cats displayed a low feed intake or refused the diet, thus these cats were also removed from the study. The calculation of the apparent nutrient digestibilities revealed a high crude fat digestibility (96.0 ± 1.26 % for diet A and 92.7 ± 1.53 % for diet B). However, the apparent digestibility of crude protein (77.0 ± 3.48 % and 73.4 ± 3.73 % for diets A and B, respectively) and of the specific amino acids was only moderate. CONCLUSION: Both diets based on larvae meal of Hermetia illucens were generally tolerated by most of the cats. However, individual differences were also present. Because the apparent crude protein digestibility was only moderate, it is recommended to consider an adequate safety margin when formulating diets based on insect protein to prevent nutrient deficiencies.

Impact of Dietary Protein Concentration and Quality on Immune Function of Cats.

Protein levels and quality in cat food can vary significantly and might affect immune function in various ways. In the present study, 3 diets with a low protein quality (LQ) and 3 diets with a high protein quality (HQ) were offered to 10 healthy adult cats for 6 weeks each, using a randomized cross-over design. The LQ and HQ diets differed in the collagen content and had low (36.7% and 36.2%), medium (45.0% and 43.3%) and high (56.1% and 54.9%) protein levels. At the end of each feeding period, blood was collected for phenotyping of leukocyte subsets, lymphocyte proliferation assay and cytokine measurements, phagocytosis assay and differential blood count. The results demonstrated no group differences for numbers of CD4+CD8-, CD4+CD8+, CD4-CD8+, MHCII+, CD21+, SWC3+ and CD14+ cells in the blood of the cats. Proliferative activity of lymphocytes when stimulated with pokeweed mitogen, Concanavalin A and Phytohemagglutinin, M form did not differ depending on the dietary protein concentration and quality. Concentrations of tumor necrosis factor alpha and interferon gamma in the supernatant of the proliferation assay were also not affected by the dietary treatment. Blood monocyte phagocytic activity was higher (P = 0.048) and cell numbers of eosinophilic granulocytes in the blood were lower (P = 0.047) when cats were fed the low protein diets. In conclusion, only a few differences in feline immune cell populations and activity depending on dietary protein supply could be detected. However, the observed increase of eosinophilic granulocytes by a higher protein intake indicates an activation of immunological mechanisms and requires further investigation.

Effects of free amino acids on cytokine secretion and proliferative activity of feline T cells in an in vitro study using the cell line MYA-1.

In vitro studies might be an interesting screening method for targeted in vivo studies in the field of immunonutrition and help to reduce and refine animal studies. As the role of amino acids for immune function of cats has not been evaluated in detail so far, the present study aimed at investigating the effects of eight different amino acids (arginine, leucine, isoleucine, valine, glutamine, lysine, threonine and tryptophan) in six concentrations each (0, 0.25, 0.5, 1, 2 and 8x the cat blood level) on cytokine secretion and proliferative activity of feline T cells (MYA-1) in vitro. The results demonstrated that high doses of arginine increased IL-4, IL-10 and TNF-α secretion of T cells, while increasing concentrations of lysine increased IL-10 secretion and proliferative activity of the T cells. High doses of leucine enhanced GM-CSF and IL-10 secretion, while concentrations of threonine in the cell culture media greater than blood concentration also increased GM-CSF and additionally TNF-α secretion of the cells. The effects of glutamine and isoleucine on T cell function were only small. In conclusion, the present in vitro study could evaluate the immunomodulating potential of specific amino acids for feline T cell function. High doses of arginine, lysine, leucine and threonine had a significant impact on cytokine secretion and proliferative activity of the T cells. Targeted in vivo studies should investigate the clinical relevance of dietary supplementation of those amino acids in healthy and diseased cats as a next step.

Impact of Increasing Dietary Calcium Levels on Calcium Excretion and Vitamin D Metabolites in the Blood of Healthy Adult Cats.

BACKGROUND: Dietary calcium (Ca) concentrations might affect regulatory pathways within the Ca and vitamin D metabolism and consequently excretory mechanisms. Considering large variations in Ca concentrations of feline diets, the physiological impact on Ca homeostasis has not been evaluated to date. In the present study, diets with increasing concentrations of dicalcium phosphate were offered to ten healthy adult cats (Ca/phosphorus (P): 6.23/6.02, 7.77/7.56, 15.0/12.7, 19.0/17.3, 22.2/19.9, 24.3/21.6 g/kg dry matter). Each feeding period was divided into a 10-day adaptation and an 8-day sampling period in order to collect urine and faeces. On the last day of each feeding period, blood samples were taken. RESULTS: Urinary Ca concentrations remained unaffected, but faecal Ca concentrations increased (P < 0.001) with increasing dietary Ca levels. No effect on whole and intact parathyroid hormone levels, fibroblast growth factor 23 and calcitriol concentrations in the blood of the cats were observed. However, the calcitriol precursors 25(OH)D2 and 25(OH)D3, which are considered the most useful indicators for the vitamin D status, decreased with higher dietary Ca levels (P = 0.013 and P = 0.033). Increasing dietary levels of dicalcium phosphate revealed an acidifying effect on urinary fasting pH (6.02) and postprandial pH (6.01) (P < 0.001), possibly mediated by an increase of urinary phosphorus (P) concentrations (P < 0.001). CONCLUSIONS: In conclusion, calcitriol precursors were linearly affected by increasing dietary Ca concentrations. The increase in faecal Ca excretion indicates that Ca homeostasis of cats is mainly regulated in the intestine and not by the kidneys. Long-term studies should investigate the physiological relevance of the acidifying effect observed when feeding diets high in Ca and P.