Negative consequences of reduced protein diets supplemented with synthetic amino acids for performance, intestinal barrier function, and caecal microbiota composition of broiler chickens.

The consequences of feeding broiler chickens with reduced protein (RP) diets for gut health and barrier function are not well understood. This study was performed to elucidate the effect of reducing dietary protein and source of protein on gut health and performance parameters. Four experimental diets included 2 control diets with standard protein levels either containing meat and bone meal (CMBM) or an all-vegetable diet (CVEG), a medium RP diet (17.5% in growers and 16.5% in finisher), and a severe RP diet (15.6% in grower and 14.6% in finisher). Off-sex Ross 308 birds were assigned to each of the 4 diets and performance measurements were taken from d 7 to 42 post-hatch. Each diet was replicated 8 times (10 birds per replicate). A challenge study was conducted on additional 96 broilers (24 birds per diet) from d 13 to 21. Half of the birds in each dietary treatment were challenged by dexamethasone (DEX) to induce a leaky gut. Feeding birds with RP diets decreased weight gain (P < 0.0001) and increased feed conversion ratio (P < 0.0001) from d 7 to 42 compared with control diets. There was no difference between CVEG and CMBM control diets for any parameter. The diet containing 15.6% protein increased (P < 0.05) intestinal permeability independent of the DEX challenge. Gene expression of claudin-3 was downregulated (P < 0.05) in birds fed 15.6% protein. There was a significant interaction between diet and DEX (P < 0.05) and both RP diets (17.5% and 15.6%) downregulated claudin-2 expression in DEX-challenged birds. The overall composition of the caecal microbiota was affected in birds fed 15.6% protein having a significantly lower richness of microbiota in both sham and DEX-injected birds. Proteobacteria was the main phylum driving the differences in birds fed 15.6% protein. At the family level, Bifidobacteriaceae, Unclassified Bifidobacteriales, Enterococcaceae, Enterobacteriaceae, and Lachnospiraceae were the main taxa in birds fed 15.6% protein. Despite supplementation of synthetic amino acids, severe reduction of dietary protein compromised performance and intestinal health parameters in broilers, evidenced by differential mRNA expression of tight junction proteins, higher permeability, and changes in caecal microbiota composition.

Effects of Caffeine and Glucose Supplementation at Birth on Piglet Pre-Weaning Growth, Thermoregulation, and Survival.

Piglet pre-weaning mortality of approximately 15% represents a major economic and welfare concern to the pork industry. Supplementing neonatal piglets with glucose and/or caffeine has the potential to counteract hypoxic stress experienced during parturition and provide an energy substrate, which may improve survival to weaning. This study investigated the effects of caffeine and glucose supplementation at birth, in combination or separately, on piglet growth, thermoregulatory ability, and pre-weaning survival. At birth, 398 piglets were assigned to one of four oral treatments: saline, glucose (300 mg), caffeine (30 mg), or caffeine and glucose combined (30 mg caffeine and 300 mg glucose), dissolved in 6 mL saline. Piglets were tagged at birth, and time taken to reach the udder was recorded. Rectal temperatures were recorded at 4 h and 24 h post-partum, and body weights recorded at birth and 1, 3, and 18 days of age. Colostrum intake was estimated using birth and day 1 weights, and all pre-weaning mortalities were recorded. Treatments did not affect rectal temperature, colostrum intake, or pre-weaning mortality (p > 0.05). Low birth weight piglets (<0.9 kg) treated with caffeine and glucose had increased growth between 1 and 3 days of age (p < 0.05) compared to low birth weight piglets of other treatment groups. Caffeine supplementation alone reduced overall pre-weaning growth in low birth weight piglets compared to all other treatments (p = 0.05). Oral caffeine and glucose had no significant effect on piglet performance except in low birthweight piglets, where it improved growth in the first 3 days of life. Caffeine and glucose supplementation in combination may be beneficial for low birth weight piglets.

Synthetic Olfactory Agonist Use in the Farrowing House to Reduce Sow Distress and Improve Piglet Survival.

The aim of the study was to investigate if the application of a synthetic olfactory agonist (SOA) would reduce indicators of stress in sows, in response to a stressor prior to parturition, and if it would improve farrowing house performance of sows and their piglets. Two studies were conducted: an intensive study with 47 sows, either having their first or second litter (Control n = 24; SOA n = 23); and a commercial validation study with 418 sows, either having their first litter or have had multiple litters (Control n = 210; SOA n = 208). Within the intensive study, sows were housed with or without a synthetic olfactory agonist suspended in the creep area of the farrowing crate, whereas within the commercial validation study, sows were housed with or without a synthetic olfactory agonist suspended over the adjoining creep area of two farrowing crates. Within the intensive study, despite a discernible increase in cortisol concentration in response to a stressor (snout rope test), cortisol response was not different between treatments (p > 0.05). Farrowing duration in first-litter sows exposed to the SOA was decreased (p < 0.001) whilst there was no impact on farrowing duration in second litter sows. Piglets were not attracted by the SOA to increase their utilisation of the creep area and spent more time in proximity to the sow (p < 0.05). Within the commercial validation study, no impacts were seen on piglet production measures (p > 0.05). Largely the use of an SOA within the farrowing house did not impact the sow or her piglets in either the intensive study or commercial validation study. Based on these current results, the use of SOA within the farrowing house is not supported.

Maternal supplementation with phytogenic additives influenced the faecal microbiota and reproductive potential in sows.

Sows undergo physiological stress during gestation and lactation, potentially leading to enteric dysbiosis and reduced reproductive potential. Phytogenic additives (PFs) may improve performance via their antioxidant, anti-inflammatory and antimicrobial properties. This study determined whether the provision of a gestation/lactation diet containing PAs would alter the gastrointestinal microbiota of sows and their piglets, and improve performance. Sows received a commercial diet throughout gestation and lactation (CTR; n = 64), a commercial diet throughout gestation and a diet containing PAs in lactation (CTR-PA; n = 63) or a commercial diet containing PAs in gestation and lactation (PA; n = 90). Sows were weighed and backfat recorded after mating and at entry and exit from the farrowing house and piglets were weighed on days 1 and 21 of life. Faecal samples collected from sows at farrowing house entry and piglets at 21 and 35 d were subjected to 16 S rRNA gene amplicon analysis. The addition of PAs to sow diets resulted in more piglets born (P = 0.03), however, it did not improve the number of liveborn piglets (P = 0.14). There were no differences in sow weight, P2 backfat depth or lactation feed intake observed. PAs had no effect on piglet weight or survival to weaning but did alter the faecal microbiota of sows, and this change was observed in piglets at 21 and 35 d. PA supplementation to sows has the potential to increase litter size, while also potentially influencing gastrointestinal tract health of the sow and piglets reared.

Exposure to maternal feces in lactation influences piglet enteric microbiota, growth, and survival preweaning.

It is known that gilt progeny performance is reduced compared with sow progeny. Previous research suggests that the presence of maternal feces in early life improves the health and survival of offspring. Therefore, we aimed to determine whether contact with feces from multiparous (MP) sows would improve the growth and survival of piglets born and reared on primiparous (P1) sows and if so, whether these differences are associated with the gut microbiota. Four treatments were applied for 10 days: Donor (n = 29) piglets had limited access to maternal feces as, each morning, sow feces were removed and placed in the crate of a P1 sow (P1-FT; n = 30 piglets) and P1-Con (n = 29) and MP-Con (n = 33) piglets had access to their own mothers' feces. All piglets were weighed on days 1, 3, 10, and 18. Fecal samples were collected from a subset of sows (n = 10/treatment) 3 days post farrow and from two female piglets/litter on days 10 and 18 (n = 20/treatment) and subject to 16S rRNA amplicon analysis. Escherichia, Clostridium, Campylobacter, and Treponema were more abundant in MP sows, while P1 sows had a higher abundance of Lactobacillus and Prevotella. At 10 days, P1 progeny fecal microbiota differed, and growth and survival were reduced when compared with MP progeny. No treatment effect was observed for P1-FT piglets (P > 0.05). Donor piglets had a different fecal microbiota and improved weight and survival then all other treatments (P < 0.05). Overall, the removal of sow feces from the farrowing crate improved piglet microbiota development, growth, and survival.

Characterisation of Early Microbial Colonisers within the Spiral Colon of Pre- and Post-Natal Piglets.

Initial enteric microbial colonisation influences animal health and disease, hence an understanding of the first microbial colonisers within the piglet is important. The spiral colon of piglets that were stillborn (n = 20), born-alive (n = 10), and born alive and had sucked (n = 9) were collected from 28 sows to investigate whether initial microbial colonisation occurs pre- or post-partum and how it develops during the first 24 h post-partum. To examine this, DNA was extracted and 16S rRNA amplicon analysis was performed to allow analysis of microbial communities. The results indicate that microbial colonisation of the spiral colon had occurred in stillborn pigs, suggesting microbial exposure prior to birth. Alpha diversity metrics indicated that the number of taxa and community richness were higher in piglets that sucked (p < 0.001) and community evenness was lower in stillborns in comparison to born-alive (p < 0.001) but was not affected by colostrum consumption (p < 0.001). Additionally, when compared with stillborn piglets, the bacteria colonising the spiral colon during the first 24 h post-partum included the potentially pathogenic bacteria Escherichia coli, Clostridium perfringens and Clostridium celatum, and potentially beneficial bacteria Lactobacillus reutueri and Faecalibacterium prausnitzii. The relative presence of Archaea was high in stillborn piglets but decreased with post-natal environmental exposure. It is evident that stillborn piglets have bacteria present within their spiral colon, however further studies are needed in order to determine the time at which colonisation is initiated and the mechanisms determining how colonisation occurs. Additionally, as expected, the immediate post-natal environment largely influences the microorganisms colonising, while colostrum consumption further contributes to the microbial community enrichment.

A Single Faecal Microbiota Transplantation Altered the Microbiota of Weaned Pigs.

Weaning is a stressful time for piglets, often leading to weight loss and is associated with increased morbidity and mortality. A leading cause for these post-weaning problems is enteric dysbiosis and methods to improve piglet health at this crucial developmental stage are needed. This study aimed to determine whether an enteric dysbiosis caused by weaning could be corrected via a faecal microbiota transplantation (FMT) from healthy piglets from a previous wean. Two or four focal piglets per litter were assigned to one of two treatments; FMT two days post weaning (n = 21; FMT) or a control which received saline two days post weaning (n = 21; CON). FMT consisted of homogenised donor faeces administered orally at 3 mL/kg. Weaning occurred at 18 days of age and weights and faecal samples were collected on days 18, 20, 24 and 35. 16S rRNA amplicon analysis was used to assess the faecal microbiota of piglets. FMT increased Shannon's diversity post weaning (p < 0.001) and reduced the scratch score observed at 24 days of age (p < 0.001). The bacterial populations significantly differed in composition at each taxonomic level. In FMT pigs, significant increases in potentially pathogenic Escherichia coli were observed. However, increases in beneficial bacteria Lactobacillus mucosae and genera Fibrobacteres and Bacteroidetes were also observed in FMT treated animals. To our knowledge, this is the first study to observe a significant effect on piglet faecal microbiota following a single FMT administered post weaning. Therefore, FMT post weaning can potentially alleviate enteric dysbiosis.

Faecal Microbiota Analysis of Piglets During Lactation.

Antimicrobial use in animals and the potential development of antimicrobial resistance is a global concern. So, non-antimicrobial techniques for animal disease control are needed. This study aimed to determine whether neonatal ceftiofur (CF) treatment affects piglet faecal microbiomes and whether faecal microbiome transplantation (FMT) can correct it. Two focal piglets per sow were assigned to treatments as follows: cffresh (n = 6) received CF (3 mg/kg intramuscular) at 7 d and fresh FMT at 13 d; cffrozen (n = 7) received CF at 7 d and frozen FMT at 13 d; CF (n = 8) received CF at 7 d and no FMT; and no CF (n = 5) received no CF or FMT. DNA was extracted from faecal samples collected on days 7, 13, and 18 for 16S rRNA amplicon analysis. All faecal blends used for the FMT consisted of pooled donor pig faeces at 1:2 ratio with saline, delivered orally at 3 mL/kg. Alpha and beta diversity metrics increased with age (p < 0.05). However, no effect of antibiotic or FMT treatment was evident in 13 and 18 d old piglets (p > 0.05). Although no effect of treatment was observed, information regarding microbial membership during lactation was gained.

Development and Function of the Intestinal Microbiome and Potential Implications for Pig Production.

The intestinal microbiota has received a lot of attention in recent times due to its essential role in the immune system development and function. Recent work in humans has demonstrated that the first year of life is the most critical time period for microbiome development with perturbations during this time being proven to have long term health consequences. In this review, we describe the literature surrounding early life events in humans and mice that contribute to intestinal microbiota development and function, and compare this to piglets predominantly during their lactation period, which focuses on the impact lactation management practices may have on the intestinal microbiota. Although extensive research has been conducted in this area in humans and mice, little research exists in pigs during perceivably the most critical time period of development, which is the lactation period. The research reviewed outlines the importance of appropriate intestinal microbiota development. However, further research is needed in order to understand the full extent routine farm practices have on a piglet's intestinal microbiota.