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The present study investigated in dogs the dietary effects of intact seaweeds on some fecal bacterial populations and metabolites, fecal IgA and apparent total tract digestibility (ATTD). Ten healthy adult dogs were enrolled in a 5 × 5 replicated Latin square design to evaluate five dietary treatments: control diet (CD), CD + Ascophyllum nodosum, CD + Undaria pinnatifida, CD + Saccharina japonica, CD + Palmaria palmata (n replicates per treatment = 10). Seaweeds were added to food at a daily dose of 15 g/kg. The CD contained silica as a digestion marker. Each feeding period lasted 28 d, with a 7 d wash-out in between. Feces were collected at days 21 and 28 of each period for chemical and microbiological analyses. Fecal samples were collected during the last five days of each period for ATTD assessment. Dogs showed good health conditions throughout the study. The fecal chemical parameters, fecal IgA and nutrient ATTD were not influenced by algal supplementation. Similarly, microbiological analyses did not reveal any effect by seaweed ingestion. In conclusion, algal supplementation at a dose of 15 g/kg of diet failed to exert noticeable effects on the canine fecal parameters evaluated in the present study.

Simple Summary The present study investigated for the first time the impact that different sampling points have on the abundance of microbial populations and metabolites within the canine stool. We found that inner stool subsamples resulted in higher concentrations of bacterial metabolites but not of microbial populations. These findings suggest that stool subsampling is unlikely to represent the canine microbiota and metabolome uniformly. We believe that complete homogenisation of the whole stool prior to analysis may improve the final outcome when investigating the canine gut microbiome. Abstract Canine faecal microbial populations and metabolome are being increasingly studied to understand the interplay between host and gut microbiome. However, the distribution of bacterial taxa and microbial metabolites throughout the canine stool is understudied and currently no guidelines for the collection, storage and preparation of canine faecal samples have been proposed. Here, we assessed the effects that different sampling points have on the abundance of selected microbial populations and bacterial metabolites within the canine stool. Whole fresh faecal samples were obtained from five healthy adult dogs. Stool subsamples were collected from the surface to the inner part and from three equally sized areas (cranial, central, caudal) along the length axis of the stool log. All samples were finally homogenised and compared before and after homogenisation. Firmicutes, Bacteroidetes, Clostridium cluster I, Lactobacillus spp., Bifidobacterium spp. and Enterococcus spp. populations were analysed, as well as pH, ammonia and short-chain fatty acids (SCFA) concentrations. Compared to the surface of the stool, inner subsamples resulted in greater concentrations of SCFA and ammonia, and lower pH values. qPCR assay of microbial taxa did not show any differences between subsamples. Homogenisation of faeces does not affect the variability of microbial and metabolome data. Although the distribution patterns of bacterial populations and metabolites are still unclear, we found that stool subsampling yielded contradictory result and biases that can affect the final outcome when investigating the canine microbiome. Complete homogenisation of the whole stool is therefore recommended.

The effect of increasing dietary doses of lactose on canine faecal microbiota and apparent digestibility was evaluated. Fourteen adult healthy dogs [1–5 years of age, mean body weight (BW) of 19.0 kg] were fed with an extruded diet containing silica (5 g/kg) as a digestion marker. After a 20 d adaptation period, increasing doses of lactose were added to the dogs’ diet (0.5, 1 and 2 g/kg BW0.75/d) during three consecutive 20-d supplementation periods. Faeces were collected at the end of each period for analyses. Four dogs refused the diet added with lactose at 0.5 g/kg BW0.75/d and were excluded from the trial, as well as two dogs, which developed acute diarrhoea when lactose was fed at 1 g/kg BW0.75/d. Conversely, eight dogs remained healthy throughout the study. Faecal moisture was influenced by lactose (quadratic, p = .001), while faecal pH and ammonia were not affected by treatments. Lactose supplementations tended to linearly decrease isovalerate (p = .051) and quadratically influence n-valerate (p = .056) in canine faeces. No changes in faecal microbial populations were observed. Apparent digestibility of dry matter, Ca, K, Mn and Fe was influenced by lactose supplementation (quadratic, p

Background Feeding dogs with diets rich in protein may favor putrefactive fermentations in the hindgut, negatively affecting the animal’s intestinal environment. Conversely, prebiotics may improve the activity of health-promoting bacteria and prevent bacterial proteolysis in the colon. The aim of this study was to evaluate the effects of dietary supplementation with fructooligosaccharides (FOS) on fecal microbiota and apparent total tract digestibility (ATTD) in dogs fed kibbles differing in protein content. Twelve healthy adult dogs were used in a 4 × 4 replicated Latin Square design to determine the effects of four diets: 1) Low protein diet (LP, crude protein (CP) 229 g/kg dry matter (DM)); 2) High protein diet (HP, CP 304 g/kg DM); 3) Diet 1 + 1.5 g of FOS/kg; 4) Diet 2 + 1.5 g of FOS/kg. The diets contained silica at 5 g/kg as a digestion marker. Differences in protein content were obtained using different amounts of a highly digestible swine greaves meal. Each feeding period lasted 28 d, with a 12 d wash-out in between periods. Fecal samples were collected from dogs at 0, 21 and 28 d of each feeding period. Feces excreted during the last five days of each feeding period were collected and pooled in order to evaluate ATTD. Results Higher fecal ammonia concentrations were observed both when dogs received the HP diets (p

The aim of the present study was to develop a new in vitro method for evaluating the digestibility of commercial diets for dogs. First, in order to develop the in vitro method, the digestibility of four commercial diets for dogs was evaluated through several in vitro trials and results were compared with those that were retrieved from the literature. The in vitro method that was developed consists of two incubation phases, a first lasting 2h and taking place in the presence of pepsin, gastric lipase and HCl (gastric phase) and a second 4h one with pancreatin and bile salts (intestinal phase). Later, digestibility of 16 extruded diets for dogs was evaluated both in vivo with adult dogs and in vitro. There was a close linear relationship between in vivo total tract and in vitro dry matter digestibility (r2 = .81), whereas accuracy of crude protein digestibility using the in vitro method was lower (r2 = .51). Linear regression accuracy for ether extract and starch digestibility was low, but the digestibility results obtained with the in vitro method (95.3 and 98.7% for ether extract and starch, respectively) were very close to those from the in vivo trial (average digestibility of ether extract and starch was 94.8 and 99.1%, respectively). The present in vitro method has proved to be a relatively simple, quick procedure for predicting the digestibility of commercial diets for dogs. The utilisation of such a method may significantly reduce the need for in vivo digestion trials with dogs.

The present study investigated the in vitro effects of tylosin (TYL), alone or associated with prebiotics (PRE), on selected canine fecal parameters. Eight treatments were set up: control diet with no addition of substrates, TYL, Fructooligosaccharides (FOS), Galactooligosaccharides (GOS), Xylooligosaccharides (XOS), TYL + FOS, TYL + GOS, TYL + XOS. The flasks (five for treatment), containing a canine fecal suspension (prepared with the feces of healthy adult dogs) and the residue of an in vitro digested dry dog food, were incubated in an anaerobic chamber at 39 &deg, C. TYL and PRE were added at a concentration of 0.2 and 1 g/L, respectively. Samples were collected after 6 and 24 h for analyses. PRE decreased pH values, iso-butyrate, and iso-valerate throughout the incubation, increased lactobacilli, cadaverine, and, tendentiously, total volatile fatty acids after 6 h, increased n-butyrate, putrescine, spermidine, and reduced spermine and E. coli after 24 h. TYL resulted in lower total volatile fatty acids and lactobacilli and higher Clostridium cluster I after 6 h and higher pH values, spermidine, and E. coli throughout the study. When associated with TYL, PRE counteracted some undesirable effects of the antibiotic such as the decrease of lactobacilli and Clostridium cluster XIVa at both 6 and 24 h. In the present study, TYL exhibited inhibitory effects on canine fecal microbiota partially counteracted by PRE supplementation.

Changing trends in fish feed formulation, with progressively higher inclusion levels of plant ingredients, are invariably introducing more fibre despite the fact that this component cannot be utilized by most fish. The effects of increasing insoluble dietary fibre level on growth, nutrient utilisation, blood parameters and gut health in European sea bass (Dicentrarchus labrax L.) were studied over a period of 117 days. Moreover, investigation on digesta transit time through gastrointestinal evacuation pattern and digesta characteristics (moisture of digesta) were studied. Five isoproteic diets were formulated to contain increasing insoluble fibre levels, neutral detergent fibre, NDF (7.2, 8.9, 11.5, 13.1 and 15.5%) derived by the inclusion of sunflower hulls and soybean hulls. No significant differences due to fibre inclusion levels were observed in final body weight, specific growth rate, feed intake, feed conversion rate, protein and lipid efficiency. No significant differences in serum total protein, glucose, triglycerides, alkaline phosphatase and inorganic phosphorous were found. All the histological sections showed normal intestinal architecture, and inflammatory and/or degenerative changes were not present in any histological section from all subjects examined. The investigation into gastrointestinal evacuation pattern revealed no significant differences between treatments, however higher dietary fibre levels seem to increase the time required to empty the stomach while the time required to empty 90% of the hindgut content was similar in all the treatments: around 46–47 h. No differences were found between diets in the moisture content of digesta along the digestive tracts. We can conclude that the different insoluble fibre levels tested in this trial have no effects on overall performances and feed efficiency in European sea bass. Results from blood biochemistry profile and histology confirm good nutritional and health status of fish under all feeding treatments. The inclusion of fibre had no influence on digesta transit time. In formulation of feed for the on-growing of European sea bass insoluble fibre derived from sunflower hulls and soybean hulls can be included at a level of up to 15.5%.

The aim of this study was to determine the presence and the level of contamination of the most important mycotoxins (deoxynivalenol, fumonisin B1 and B2, aflatoxin B1, B2, G1 and G2, ochratoxin A and zearalenone) in 48 samples of extruded dry dog food found in the Italian market (24 samples from standard economy lines, 24 of premium lines). Analyses were performed using ultra-performance liquid chromatography coupled to tandem mass spectrometry. Although the concentrations of the mycotoxins in all samples proved to respect the European legislation with regards to animal feed, the analyses revealed a substantial presence of deoxynivalenol, fumonisins and ochratoxin A, with values above the limit of quantification (5 µg/kg) in 100%, 88% and 81% of the samples, respectively. In contrast, aflatoxins and zearalenone contamination proved to be very modest, with 88% and 75% of the samples, respectively, showing concentrations below the corresponding limit of quantification (5 µg/kg for aflatoxins and 10 µg/kg for zearalenone). Moreover, despite a very heterogeneous contamination, the concentration of fumonisins and ochratoxin A was significantly higher in standard foods than in premium ones (491 vs. 80.2 µg/kg dry matter for fumonisin B1; 113 vs. 38.5 µg/kg dry matter for fumonisin B2; 599 vs. 103 µg/kg dry matter for total fumonisins; 23.8 vs. 13.0 µg/kg dry matter for ochratoxin A; P < 0.001). Furthermore, a simultaneous presence of different mycotoxins (at concentrations higher than their limit of quantification) was observed in most of the pet foods analyzed; in particular, 19% of the samples were contaminated by no fewer than two different types of mycotoxins, 52% by three, 25% by four and 2% by all the mycotoxins evaluated. These results revealed the need for further investigation into the potential risk deriving from chronic exposure to low doses of the different types of mycotoxins that pet species are subject to today.

The digestibility (in vitro), toxicity and metabolic effects of rapeseed (RPH) and sunflower (SPH) protein hydrolysates have been evaluated in a murine animal model. The enzyme AlcalaseVR was employed to obtain a mild enzymatic hydrolysis of rapeseed and sunflower defatted seed meals (DSM) protein isolates. Both hydrolysates showed higher in vitro digestibility than the respective DSM, presumably as a consequence of the hydrolysis process that they had undergone. In vivo, RPH and SPH were well tolerated. Body and organ weights, biochemical blood parameters from treated male mice were comparable to controls. Food intake was regular in RPH and SPH animals, suggesting a good palatability of the hydrolysates. Not relevant perturbations of the principal hepatic and renal drug metabolism enzymes were observed in RPH or SPH mice. In conclusion, protein hydrolysates from sunflower and rapeseed DSM did not determine relevant toxicological effects; therefore, they could be considered as alternative protein sources and/or food ingredients.

The in vitro effect of a Yucca schidigera extract (YSE) and tannins from chestnut wood on composition and metabolic activity of canine and feline faecal microbiota was evaluated. Four treatments were carried out: control diet, chestnut tannins (CT), YSE and CT + YSE. The YSE was added to canine and feline faecal cultures at 0.1 g/l, while CT were added at 0.3 g/l for a 24-h incubation. A total of 130 volatile compounds were detected by means of headspace-solid phase microextraction gas-chromatography/mass spectrometry analyses. Several changes in the metabolite profiles of fermentation fluids were found, including a decrease of alcohols (-19%) and esters (-42%) in feline and canine inoculum, respectively, which was due to the antibacterial properties of tannins. In canine inoculum, after 6 h, YSE + CT caused lower cadaverine concentrations (-37%), while ammonia (-4%) and quinolone (-27%) were reduced by addition of CT. After 24 h, the presence of CT resulted in a decrease of sulphur compounds, such as dimethyl sulphide (-69%) and dimethyl disulphide (-20%). In feline faecal cultures, after 6 h, CT lowered the amount of indole (-48%), whereas YSE tended to decrease trimethylamine levels (-16%). Both in canine and feline inoculum, addition of CT and, to a minor extent, YSE affected volatile fatty acids patterns. In canine faecal cultures, CT exerted a marginal inhibitory effect on Escherichia coli population (-0.45 log 10 numbers of DNA copies/ml), while enterococci were increased (+2.06 log 10 numbers of DNA copies/ml) by YSE. The results from the present study show that YSE and tannins from chestnut wood exert different effects on the composition and metabolism of canine and feline faecal microbiota. In particular, the supplementation of YSE and tannins to diets for dogs and cats may be beneficial due to the reduction of the presence of some potentially toxic volatile metabolites in the animals' intestine.

Background The present in vitro study investigated whether the utilization of fructooligosaccharides (FOS) may influence canine fecal microbial population in presence of diets differing in their protein content and digestibility. Fresh fecal samples were collected from five adult dogs, pooled, and incubated for 24 h with the undigested residue of three diets: 1, Low protein high digestibility diet (LP HD, crude protein (CP) 229 g/kg); 2, High protein high digestibility diet (HP HD, CP 304 g/kg); 3, High protein low digestibility diet (HP LD, CP 303 g/kg) that had been previously subjected to enzymatic digestion. In the in vitro fermentation study, there were six treatments: 1) LP HD; 2) HP HD 3) HP LD; 4) LP HD + FOS; 5) HP HD + FOS; 6) HP LD + FOS. Fructooligosaccharides were added at the final concentration of 1.5 g/L. Samples of fermentation fluid were collected at 6 and 24 h of incubation. Results Values of pH were reduced by FOS at 6 and 24 h (P

To attain a good level of animal welfare, pigs require a sufficient nvironmental illumination. Therefore, minimum levels for light duration and light intensity have been set up by the European legislation (Directive 2008/120). An experimental trial was designed to determine whether an increased duration of the photophase (up to 16 h of light per day) could modify the behavior, productive parameters, and meat and ham quality of Italian heavy pigs. Forty crossbred (Large White × Landrace) castrated males pigs (26 kg initial average BW) intended for Protected Designation of Origin (according to European Union Regulation - European Union, 2012) dry-cured ham production were raised according to Parma ham production rules up to the weight of 160 kg. Pigs were homogeneously allotted to 2 experimental groups, each comprising 20 pigs. The short photoperiod (SP) group received the minimum mandatory number of hours of light per day (corresponding to 8 h/d), whereas the long photoperiod (LP) group was subjected to 16 h of light per day during the whole production cycle. Light intensity was maintained at 40 lux (i.e., the minimum mandatory level) for both the experimental groups. Growth and slaughtering parameters, carcass traits, fatty acid composition, meat and dry-cured ham quality, and animal behavior were assessed. Pigs in the LP group showed a greater live weight and carcass weight compared to the SP group (P = 0.005 and P = 0.007, respectively). Similarly, hams obtained from the LP group were significantly heavier and their weight losses during the dry-curing period were reduced (P < 0.01) when compared to the SP group. No significant differences were detected between the experimental groups as concerns meat and ham quality or fatty acid composition of the subcutaneous fat. Pigs in the LP group spent more time resting and less time pseudo-rooting (P < 0.01). Our results indicate that, given an appropriate dark period for animal rest, an increased duration of the photoperiod, even at the lower mandatory light intensity level, can favorably affect growth parameters of heavy pigs without any negative effect on animal behavior, carcass traits, or meat or long-cured ham quality. Therefore, rearing pigs in semidarkness should be considered as a baseless practice, contrary to animal welfare.

Obesity represents the most common nutritional pathology in companion animals, with an estimated prevalence of overweight or obese dogs of at least one third of the canine population. Aim of the present study was to evaluate the nutritional status of a canine population in the USA and assess its correlation with some specific individual factors. During a period of about two months, data were collected concerning nutritional status, dietary management and clinical situation of a group of 158 adult dogs. Half of the animals resulted overweight or obese and the average BCS was 3.5 (on a 5-points scale). Factors such as age, neutering, hormone and chronic arthritic disorders, as well as the belonging to specific breeds showed a positive significant correlation with BCS. On the contrary, gender, presence of other animals and type of food offered did not show any correlation with BCS. Despite the relatively low number of dogs involved, present results confirm that overweight affects a large portion of the canine population in the USA and is influenced mainly by endogenous factors.

Objective—To evaluate the growth kinetics of a strain of Bifidobacterium pseudocatenulatum (BP) on 4 oligo- or polysaccharides and the effect of feeding a selected probiotic-prebiotic combination on intestinal microbiota in cats. Animals—10 healthy adult cats. Procedures—Growth kinetics of a strain of cat-origin BP (BP-B82) on fructo-oligosaccharides, galacto-oligosaccharides (GOS), lactitol, or pectins was determined, and the combination of GOS and BP-B82 was selected. Cats received supplemental once-daily feeding of 1% GOS–BP-B82 (1010 CFUs/d) for 15 days; fecal samples were collected for analysis the day before (day 0) and 1 and 10 days after the feeding period (day 16 and 25, respectively). Results—Compared with the prefeeding value, mean fecal ammonia concentration was significantly lower on days 16 and 25 (288 and 281 μmol/g of fecal dry matter [fDM], respectively, vs 353 μmol/g of fDM); fecal acetic acid concentration was higher on day 16 (171 μmol/g of fDM vs 132 μmol/g of fDM). On day 16, fecal concentrations of lactic, n-valeric, and isovaleric acids (3.61, 1.52, and 3.55 μmol/g of fDM, respectively) were significantly lower than on days 0 (5.08, 18.4, and 6.48 μmol/g of fDM, respectively) and 25 (4.24, 17.3, and 6.17 μmol/g of fDM, respectively). A significant increase in fecal bifidobacteria content was observed on days 16 and 25 (7.98 and 7.52 log10 CFUs/g of fDM, respectively), compared with the prefeeding value (5.63 log10 CFUs/g of fDM). Conclusions and Clinical Relevance—Results suggested that feeding 1% GOS–BP-B82 combination had some positive effects on the intestinal microbiota in cats

The aim of the present study was to evaluate the effect on the composition and activity of the canine intestinal microbiota of different sources of soluble fibre that are among the most widely used by the pet food industry, and some prebiotic substances that might be considered as potential ingredients for dog diets. In vitro, there were 14 treatments: Control diet; Lactitol; Beneo P95 (FOS from partially hydrolyzed inulin from chicory); Fibrulose F97 (FOS from chicory); Fibruline XL (inulin from chicory); Pectin Classic AU202 (pectins from apple); Pectin Classic CU201 (pectins from citrus fruit); Beneo GR (inulin from chicory); Gluconic acid; Dried chicory; Dried beet pulp; Exafine 500 (pea hull fibre); Psyllogel (850 g/kg Psyllium fibre); Benefibra (partially hydrolyzed guar gum). Substrates were added at 1 g/L (treatments 2 through 9) or 4 g/L (treatments 10 through 14) to canine faecal cultures for a 24 h incubation. In view of the different beneficial properties that they showed during the in vitro trial, Lactitol and Pectin Classic CU201 were selected to be tested in a feeding study with 16 privately owned dogs. Dogs were divided into two treatment groups of eight animals each; all dogs were fed the same commercial dry diet (the same one used for the in vitro study) and received 1) lactitol or 2) Pectin Classic CU201 at 10 g/kg of diet for 30 d. Faecal samples were collected at 0, 20, and 30 d after the start of the trial and 10 d (Day 40) after prebiotic withdrawal. In vitro, compared with the control, gas production and maximum rate of gas production were increased (P0.05) by treatment, whereas prebiotics resulted in reduced concentrations of coliforms and C. perfringens (P