Leticia Abecia

Study of the effect of presence or absence of protozoa on rumen fermentation and microbial protein contribution to the chyme.

The aim of this study was to investigate the effect of presence or absence of protozoa on rumen fermentation and efficiency of microbial protein synthesis under different diets. Of 20 twin paired lambs, 1 lamb of each pair was isolated from the ewe within 24 h after birth and reared in a protozoa-free environment (n = 10), whereas their respective twin-siblings remained with the ewe (faunated, n = 10). When lambs reached 6 mo of age, 5 animals of each group were randomly allocated to 1 of 2 experimental diets consisting of either alfalfa hay as the sole diet, or 50:50 mixed with ground barley grain according to a 2 × 2 factorial arrangement of treatments. After 15 d of adaptation to the diet, the animals were euthanized and total rumen and abomasal contents were sampled to estimate rumen microbial synthesis using C(31) alkane as flow marker. Different ((15)N and purine bases) and a novel (recombinant DNA sequences) microbial markers, combined with several microbial reference extracts (rumen protozoa, liquid and solid associated bacteria) were evaluated. Absence of rumen protozoa modified the rumen fermentation pattern and decreased total tract OM and NDF digestibility in 2.0 and 5.1 percentage points, respectively. The effect of defaunation on microbial N flow was weak, however, and was dependent on the microbial marker and microbial reference extract considered. Faunated lambs fed with mixed diet showed the greatest rumen protozoal concentration and the least efficient microbial protein synthesis (29% less than the other treatments), whereas protozoa-free lambs fed with mixed diet presented the smallest ammonia concentration and 34% greater efficiency of N utilization than the other treatments. Although (15)N gave the most precise estimates of microbial synthesis, the use of recombinant DNA sequences represents an alternative that allows separate quantification of the bacteria and protozoa contributions. This marker showed that presence of protozoa decrease the bacterial-N flow through the abomasum by 33%, whereas the protozoa-N contribution to the microbial N flow increased from 1.9 to 14.1% when barley grain was added to the alfalfa hay. Absolute data related to intestinal flow must be treated with caution because the limitations of the sampling and maker system employed.

Rumen microbial population dynamics in response to photoperiod

Aims:  This work was carried out to determine if there was a difference in the microbial population of the rumen associated with daylength at which sheep are housed.

The effect of lactating rabbit does on the development of the caecal microbial community in the pups they nurture

Aims:  To study the effect of microbial community of the rabbit does as influenced by dietary factors, on the development of the gut microbiota of their litters.

The effect of medicated diets and level of feeding on caecal microbiota of lactating rabbit does

Aims:  To study the effect of the type of antibiotic used in medicated diets against pathogens and the feeding level on the microbial biodiversity in the rabbit caecum.

Rumen protozoal diversity in the Spanish ibex (Capra pyrenaica hispanica) as compared with domestic goats (Capra hircus)

Rumen protozoal diversity in the Spanish ibex (SI) was studied in males (n=4), females (n=7) and young (n=4) from the Maestrazgo (Spain) and contrasted with domestic goats (n=3; DG) of the same region. There were no differences among SI types in protozoal concentration or in the number of protozoal species. Only protozoa from the genus Entodinium were observed in SI (seven species), the highest numbers corresponding to E. damae, E. ovibos and E. parvum. DG harboured threefold more species than SI. Nine to 10 Entodinium spp. were observed, but E. ovibos was absent from the rumen of DG, and E. damae was in only one animal. E. caudatum (caudatum, dubardi and lobospinosum morphotypes) occurred in the highest percentage, and E. dubardi, E. exiguum and E. parvum were quite abundant. Four genera of the subfamily Diplodiniinae and the genera Isotricha and Dasytricha from the family Isotrichidae were detected in DG. Epidinium (two DG) and Ophryoscolex (one DG) were also observed. Denaturing gradient gel electrophoresis analysis agreed with microscopic classification, showing up to 8 and 16 bands in SI and DG samples, respectively. The three DG clustered together (similarity index over 0.84), and separately from SI (similarity index over 0.86), with only 0.58 similarity between host species.

Biodiversity and fermentative activity of caecal microbial communities in wild and farm rabbits from Spain.

In order to study the microbial caecal ecosystem of wild and domestic rabbits through the fermentation characteristics and concentration and diversity of bacterial and archaeal communities, caecal samples from sixteen wild rabbits (WR) were contrasted with two groups (n = 4) of farm rabbits receiving low (LSF) or high (HSF) soluble fibre diets from 28 (weaning) to 51 days of age. DNA was extracted for quantifying bacteria and Archaea by qPCR and for biodiversity analysis of microbial communities by DGGE. Samples from WR had lower caecal pH and ammonia and higher volatile fatty acids concentration than farm animals. Lower acetate and higher butyrate proportions were detected in WR. Bacterial and archaeal DGGE profiles were clearly different between wild and farm rabbits, and diet-affected population of farm rabbits. Similarity index of bacteria was lower than 0.40 among WR, and 0.52 among farm rabbits. In conclusion, caecal fermentation characteristics differ between wild and farm rabbits, which harbour clearly different bacterial and archaeal communities. In farm rabbits, diversity is influenced by the dietary level of soluble fibre.

Bacterial profile from caecal contents and soft faeces in growing rabbits given diets differing in soluble and insoluble fibre levels

To verify if non-invasive collection of soft faeces (SF) from rabbits can be used as an index of bacterial biodiversity in caecal contents (CC), 24 weaned rabbits were given diets with low (LI) and high (HI) levels of insoluble fibre (neutral detergent fibre, NDF) and low (LS) and high (HS) levels of soluble fibre (neutral detergent soluble fibre, NDSF). After 21 days, animals were fitted with neck collars for SF collection. Two days later, animals were slaughtered and CC sampled. Total bacterial concentration quantified by real time PCR (log(10) ng DNA/mg DM) was higher in SF than CC (2.615 vs. 2.383). Among diets, in CC it was (P = 0.059) lowest in LILS diet, whereas in SF it decreased (P = 0.025) with the NDF level. DGGE profiles showed that structure of bacterial communities of SF was close to that of CC; however, similarity was higher in LI than HI diets (0.82 vs. 0.74). Diversity indexes in CC decreased with NDSF (P < 0.05), whereas the effect of NDF (P < 0.05) was also appreciated in SF. Soft faeces can be an alternative to surgery or slaughter techniques to monitor changes in caecal bacterial community; however, high dietary NDF may decrease similarity between both communities.

Contribution of gut microbial lysine to liver and milk amino acids in lactating does

The contribution of microbial amino acids through caecotrophy to tissue protein metabolism was investigated in lactating does. Attempts were made to vary microbial supply through a dietary antibiotic, Zn bacitracin, and to vary tissue demand through manipulation of litter size. Three groups of eight New Zealand does were fed different experimental diets from day 28 of pregnancy to day 26 of lactation. The control group received the basal diet formulated to meet requirements with grass hay, wheat, soybean meal and barley grain. The second (no antibiotic) group and the third (bacitracin; BAC) group ingested the basal diet supplemented with ammonium sulfate (5 g/kg), initially unlabelled (day 1 to day 8) then labelled with 15N (day 9 to day 30), while the BAC diet was also supplemented throughout with antibiotic (Zn bacitracin; 100 mg/kg). From just after birth each group of does was subdivided into two groups, each of four females, with the litter size either five (LS5) or nine (LS9) pups. The 15N enrichment in liver, milk and caecal bacteria amino acids was determined by GC-combustion-isotope ratio MS. All amino acids in bacterial protein were enriched with the (15 NH 4)2SO4 treatment, with lysine 15N enrichment significantly greater in caecal bacteria (0.23 (SE 0.0063) atom % excess (ape)) than in liver (0.04 (SE 0.0004) ape) or milk protein (0.05 (SE 0.0018) ape), confirming the double origin (bacterial and dietary) of tissue lysine. The contribution of microbes to tissue lysine was 0.23 (SE 0.006) when milk protein was used as reference.

Effect of dietary fat to starch content on fecal microbiota composition and activity in dogs

Dietary fat is known to modulate the hindgut microbiota in rodents; however, there is no clear evidence on the impact of high-fat diets on canine gut microbiota. The purpose of this study was to investigate the effect of feeding of diets differing in the amount of ME provided by fat and starch on the composition and activity of canine fecal microbiota. Twelve adult (3 to 7 yr of age) spayed Beagle dogs received a low-fat-high-starch diet (LF-HS; approximately 23%, 42%, and 25% ME provided by fat, starch, and CP, respectively) and a high-fat-low-starch diet (HF-LS; approximately 43%, 22%, and 25% ME provided by fat, starch, and CP, respectively) following a 2-period crossover arrangement. The higher amount of fat in the HF-LS diet was provided by lard, whereas the higher amount of starch in the LF-HS diet was provided primarily by maize and broken rice. Each period lasted 7 wk and included 4 wk for diet adaptation. Dogs were fed to meet their daily energy requirements (set at 480 kJ ME/kg BW0.75). Fecal samples were collected on weeks 5 and 6 of each period for the analysis of bacterial richness, diversity, and composition [by Ion-Torrent next-generation sequencing], bile acids, ammonia, and VFA. Additional fecal samples were collected from four dogs per diet and period to use as inocula for in vitro fermentation using xylan and pectin as substrates. Gas production was measured at 2, 4, 6, 9, 12, and 24 h of incubation. On week 7, blood samples were collected at 0- and 180-min postfeeding for the analysis of bacterial lipopolysaccharide (LPS). Feeding the HF-LS diet led to a greater (P < 0.05) fecal bile acid concentration compared with the LF-HS diet. Bacterial richness and diversity did not differ between diets (P > 0.10). However, dogs showed a lower relative abundance of Prevotella (P < 0.01), Solobacterium (P < 0.05), and Coprobacillus (P ˂ 0.05) when fed of the HF-LS diet. Fecal ammonia and VFA contents were not affected by diet (P > 0.10). Relative to the LF-HS diet, in vitro fermentation of xylan using feces of dogs fed the HF-LS diet produced less gas at 6 h (P < 0.01) and 9 h (P < 0.05). Blood LPS did not increase at 180-min postfeeding with either diet (P < 0.10). These findings indicate that feeding a HF-LS diet to dogs does not affect bacterial diversity or fermentative end products in feces, but may have a negative impact on Prevotella and xylan fermentation.

Effect of litter size and bacitracin administration on tissue protein synthesis of lactating rabbit does

Bacitracin is an antibiotic used in rabbit husbandry to control microbial digestive pathologies. Collateral effects on absorption and mucosal development have been reported and these may impact on protein metabolism. This study aims to analyse the effect of the antibiotic on protein synthesis in lactating does because mammary gland metabolism and milk output should provide a sensitive index of any undesirable action of bacitracin. Rates of protein synthesis were measured in mammary gland, liver, intestinal mucosa and muscle of lactating rabbits does by injecting a flooding dose of [(2)H(5)]phenylalanine into the auricular artery of two groups (each n = 8) of New Zealand White does fed different experimental diets. The control group (C) received the basal diet and the bacitracin group (B) ingested the same diet but supplemented with bacitracin (100 mg/kg). Animals received the experimental diet from day 28 of pregnancy until day 26 of lactation when they were slaughtered. Just after birth, litter size was adjusted by cross-fostering either to five or nine pups (four does per dietary treatment). The relative weight of the liver tended to be greater in those females receiving the B diet (27 v. 22.5 g/kg BW; P < 0.07), while diet did not affect mammary gland weight (255.7 ± 10.59 g). Fractional protein synthesis rate (FSR) was higher for intestinal mucosa (duodenum; 51.7% ± 2.09%/day) followed by mammary gland and liver (38.29 ± 2.62%/day and 40.2 ± 1.98%/day, respectively), and the lowest value was observed in muscle (2.92 ± 0.26%/day; P < 0.0001). Bacitracin treatment lowered FSR in the mammary gland by 23% (P = 0.024) and this was independent of litter size. Conversely, FSR in the duodenum was not affected by antibiotic treatment but reduced by 15% (P = 0.021) for the larger litter size.