Seaweed Supplementation Failed to Affect Fecal Microbiota and Metabolome as Well as Fecal IgA and Apparent Nutrient Digestibility in Adult Dogs.

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.

On the Variability of Microbial Populations and Bacterial Metabolites within the Canine Stool. An in-Depth Analysis.

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.

In Vitro Evaluation of the Effects of Tylosin on the Composition and Metabolism of Canine Fecal Microbiota.

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 °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.

Occurrence of Mycotoxins in Extruded Commercial Cat Food.

The occurrence of the most important mycotoxins (deoxynivalenol, fumonisin B1 and B2, aflatoxins B1, B2, G1, and G2, ochratoxin A, zearalenone, T-2, and HT-2 toxins) was determined in 64 extruded cat foods purchased in Italy through ultra-performance liquid chromatography coupled with tandem mass spectrometry. Deoxynivalenol and fumonisins were the most common contaminants (quantified in 80 and 95% of the samples, respectively). Conversely, aflatoxins B2, G1, and G2 were not identified in any sample. Some cat foods exceeded the regulatory limit for aflatoxin B1 (n = 3) or the guidance values for zearalenone (n = 3), fumonisins (n = 2), ochratoxin A (n = 1), and T-2 (n = 1) recently established for pets in the European Union. A widespread co-occurrence of mycotoxins was observed (28, 42, and 8% of the samples contained quantifiable amounts of two, three, and four mycotoxins, respectively). This study describes criticisms regarding the mycotoxin issue in pet food and suggests an improvement of the monitoring of the pet food chain.

Influence of dietary protein and fructooligosaccharides on fecal fermentative end-products, fecal bacterial populations and apparent total tract digestibility in dogs.

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 < 0.001) and the supplementation with FOS (p < 0.05). The diets containing FOS resulted in greater ATTD of DM, Ca, Mg, Na, Zn, and Fe (p < 0.05) while HP diets were characterized by lower crude ash ATTD (p < 0.05). Significant interactions were observed between FOS and protein concentration in regards to fecal pH (p < 0.05), propionic acid (p < 0.05), acetic to propionic acid and acetic + n-butyric to propionic acid ratios (p < 0.01), bifidobacteria (p < 0.05) and ATTD of CP (p < 0.05) and Mn (p < 0.001). CONCLUSIONS: A relatively moderate increase of dietary protein resulted in higher concentrations of ammonia in canine feces. Fructooligosaccharides displayed beneficial counteracting effects (such as increased bifidobacteria) when supplemented in HP diets, compared to those observed in LP diets and, in general, improved the ATTD of several minerals.

An in vitro evaluation of the effects of a Yucca schidigera extract and chestnut tannins on composition and metabolic profiles of canine and feline faecal microbiota.

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.

In vitro influence of dietary protein and fructooligosaccharides on metabolism of canine fecal microbiota.

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 < 0.001); conversely, low protein digestibility and high dietary protein level resulted in higher pH at both sampling times (P < 0.001). At 24 h, FOS lowered ammonia (-10%; P < 0.001) and resulted (P < 0.05) in higher concentrations of total volatile fatty acids (VFA) (+43%), acetic acid (+14%), propionic acid (+75%) and n-butyric acid (+372%). Conversely, at 24 h, low protein digestibility resulted (P < 0.01) in lower concentrations of acetic acid (-26%), propionic acid (-37%) and total VFA (-21%). Putrescine concentrations were increased at 6 and 24 h of fermentation by low protein digestibility (+21 and 22%, respectively; P < 0.05) and FOS (+18 and 24%, respectively; P < 0.01). After 24 h of fermentation, high dietary protein level resulted in lower counts of lactobacilli and enterococci (-0.5 and -0.7 log cells/mL, respectively; P < 0.05) whereas low protein digestibility tended to increase counts of C. perfringens (+0.2 log cells/mL; P = 0.07). CONCLUSIONS: Results from the present study showed that diets rich in protein may exert negative influences on the canine intestinal ecosystem, slightly increasing the presence of ammonia and reducing counts of lactobacilli and enterococci. Moreover, the presence of poorly digestible protein resulted in lower concentrations of VFA. Conversely, administration of FOS may improve metabolism of canine intestinal microbiota, reducing ammonia concentrations and enhancing VFA production.

Effect of feeding a selected combination of galacto-oligosaccharides and a strain of Bifidobacterium pseudocatenulatum on the intestinal microbiota of cats.

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 (10(10) 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 log(10) CFUs/g of fDM, respectively), compared with the prefeeding value (5.63 log(10) 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.

Meat and ham quality of Italian heavy pigs subjected to different illumination regimes.

In order to attain a good level of welfare pigs require a sufficient environmental illumination.Therefore minimum levels for light duration and light intensity are set up by the European legislation.Two independent and separate trials were aimed to determine whether an increase above the minimum mandatory levels of lighting duration (14 vs. 8 hours/day at 70 lux: trial A) or of light intensity (80 vs. 40 lux over a 12-hour lighting period: trial B) could modify the carcass traits and meat and cured hams quality of Italian heavy pigs (body weight at slaughtering of about 160 kg). Slaughtering parameters, fresh meat quality and fatty acid composition of raw thighs and cured hams were assessed. Pigs receiving the longer photoperiod showed a tendency (P < 0.1) toward a higher slaughtering body weight and a higher saturation degree (P < 0.01) of subcutaneous fat of the raw thighs, while cured hams deriving from pigs subjected to the higher illumination intensity showed a higher (P < 0.05) relative content of polyunsaturated fatty acids. Our results indicate that, within a moderate range of light intensity and given an appropriate dark period for animal rest, neither a prolonged photoperiod nor a higher light intensity caused any negative effect on the carcass traits, meat or quality of long-cured hams.

The effects of two different light intensities on the production and behavioural traits of Italian heavy pigs.

Eighty Italian heavy pigs having an initial body weight of about 75 kg were allotted to two experimental groups: a low light intensity group (LL), in which pigs received the lowest level of illumination mandatory in piggeries (40 lux) and a high light intensity group (HL), in which pigs were exposed to a higher level of illumination (80 lux). For both groups the light phase lasted 12 hours per day. Pigs were slaughtered at about 160 kg body weight. Light intensity showed to have no effects on the growth traits, carcass parameters or neutrophil-to-lymphocyte ratio of heavy. With the exception of drinking (more frequently observed among LL pigs; P < 0.01) and sternal recumbency (more frequently observed among pigs belonging to HL group; P < 0.01), the behavioural observations, carried out on 20 pigs per group, indicated that light intensity did not affect the main behavioural traits of pigs. Agonistic interactions were significantly higher (P < 0.05) among pigs receiving a lower level of illumination (40 lux; LL group) and pigs exposed to the higher level of illumination (80 lux; HL group) showed a tendency (P < 0.1) towards a higher proportion of tactile social interactions. Our results confirm that a proper illumination level can be included among environmental conditions having a positive effect in order to preserve swine welfare.