The influence of the genetic background and feeding strategies on the health, performance and gastrointestinal homeostasis of weaned pigs

In pig intensive production system, weaning is generally considered the most stressful phase and represents a main challenge. Piglets have to face several stress factors that coupled with the lack of a complete immune competence contribute to the occurrence of post-weaning diarrhoea (PWD) syndrome mainly supported by enterotoxigenic Escherichia coli (ETEC) F4 and F18. In this phase, the host-microbial interplay plays a key role and it can be influenced both by external (i.e. diet) and internal (i.e. age, sex and genetics) factors. The present thesis reports three studies focused on genetic and feeding strategies influences on the host-microbial interplay in weaned pig through the application of new omics approach. The first study demonstrates that the pigs' Mucine 4 (MUC4) genotype (DQ848681:g.8227C>G), previously associated with ETEC F4ac susceptibility, influences the degree of jejunum fucosylation, the expression of genes involved in the intestinal homeostasis and the blood metabolic profile of healthy piglets. The first and the second studies showed that the pigs' Alpha (1,2)-fucosyltransferase (FUT1) genotype (M307 A>G), previously associated to ETEC F18 susceptibility, affects the intestinal fucosylation, the jejunal microbial balance by increasing the non-beneficial bacteria in susceptible genotypes and the blood metabolic profile of healthy piglets during pre and post-weaning. The third study evaluates the effectiveness of Saccharomyces cerevisiae (Sc) supplementation (5x1010 CFU/kg) to counteract the ETEC F4ac infection in MUC4 susceptible weaned pigs. MUC4 genotype confirmed its efficacy as a marker for ETEC F4ac susceptibility. The preventive and competitive supplementation of Sc was effective in containing the ETEC F4ac infection as showed by jejunal transcriptomic profile. The thesis provides new insight on the interplay between host-genetics, gut microbiota and host physiological status. Furthermore, it highlights that known genetic markers for ETEC F4 and F18 could be applied to stratify the animals in trials both in healthy or challenge-based protocols

Importance of Breed, Parity and Sow Colostrum Components on Litter Performance and Health

The aims of this study were to investigate the effect of breed and parity on colostrum components, and to associate sow breed, parity, and colostrum components with survival, growth, and the occurrence of diarrhoea of their litters. In Experiment 1, 64 sows (Duroc = 13; Landrace = 17 and Large White = 34) were included. In Experiment 2, 71 sows with different parities (1 = 10; 2 = 16; 3 = 13; 4 = 12; ≥5 = 20) were included. The number (N) of live piglets, litter body weight (Experiment 1), and the occurrence of diarrhoea (Experiment 1) were recorded at farrowing, at 2–3 days of age, and at weaning. Colostrum was analysed for proximate composition, immunoglobulins (Igs), and somatic cell count (SCC). Stepwise regressions and ANOVA models were used to associate breed, parity, and colostrum components with litter performance. The Duroc breed had the highest IgG and IgA (p < 0.005). Gilts had a higher fat% and SCC (p< 0.0001); these compounds were positively correlated (r = 0.45). Increased IgA tended to increase the N of weaned piglets (p = 0.058) and reduce litter diarrhoea (p = 0.021). The SCC increased the N of weaned piglets (p = 0.031). Overall, this study confirmed that breed and parity can influence the colostrum composition and highlighted the key role of Igs and somatic cells in piglet health

Contribution of L-Arginine supplementation during gestation on sow productive performance and on sow microbial faecal profile

Arginine plays an important role during reproduction, however, the amount of supplementation in sow diet is still uncertain. The aim of the study was to verify the efficacy of a gestating diet enriched or not with a low dose of L-arginine (Arg) on sow productive performance in terms of numbers and weight of piglets at birth and at weaning, frequency of intrauterine growth retardation (IUGR) and piglets' mortality, sow placenta weight and salivary humoral immunity and intestinal microbial balance of the sows. 205 sows (Landrace x Large White) were divided into two experimental groups: a control group (CON) (102 sows) and a group supplemented with 0.25% of Arg (ARG) for the whole pregnancy period. Saliva and faecal samples were collected two days before farrowing and used for immunoglobulins and microbial analysis, respectively. Arg improved the number of total born piglets (p = .043) and tended to improve the number of total born alive (p = .086) and to reduce IUGR % (p = .090) and dead piglets at d0–d3 (p = .088). The weight of placenta and humoral immunity were not influenced by Arg. Arg did not modify the faecal microbial structure (alpha and beta indices) but increased the relative abundance of Bacteroidaceae family and Bacteroides genera (p = .0001). The results support the knowledge that Arg plays a key role in nutrition and physiology of pregnant sows without compromising gut eubiosis.HighlightArg supplementation of sows' gestation diet increased the number of total bornsArg supplementation of sows' gestation diet did not negatively affect the sows' gut eubiosisArg plays a significant role in the nutrition of pregnant sows. Arg supplementation of sows' gestation diet increased the number of total borns Arg supplementation of sows' gestation diet did not negatively affect the sows' gut eubiosis Arg plays a significant role in the nutrition of pregnant sows

Pattern of antibiotic consumption in two Italian production chains differing by the endemic status for Porcine Reproductive and Respiratory Syndrome

The aim of this case study was to quantify antibiotic (AB) use in Italian weaning (W) and fattening (F) units differentiated for porcine reproductive and respiratory syndrome (PRRS) occurrence. Farms were classified as either PRRS negative (–) or PRRS positive (+) based on the circulation of the virus among the animals. In all the farms, the modified live PRRS virus (PRRSV) vaccine was provided to all the animals. In the PRRS– farms, the level of circulating antibodies was low, and the disease, in its clinical form, did not occur. In the PRRS+ farms, the level of circulating antibodies against the virus was high, and the disease was recurrent. Data regarding AB consumption were collected from 2017 to 2020, and the active compounds (ACs) were expressed as milligrams of AC/total kilogram of body weight (BW) produced. Each AC was classified into one of four categories according to the EuropeanMedicines Agency classification of ABs for prudent and responsible use in animals: Avoid, Restrict, Caution, and Prudence. Data regarding the ACs in each category were analyzed using a linear model that included production phase, PRRS status, and their interaction as factors. Performance parameters, average age of the pigs at the end of each phase, daily live weight gain, feed-to-gain ratio, total losses, cost index, and medication costs were significantly influenced by the PRRS chain. The use of class B ABs was not affected by production phase or PRRS status. Conversely, for class C ABs, interaction between the two factors (p = 0.02) was observed; W/PRRS+ and F/PRRS+ showed the greatest AB use for this class (p = 0.003). For class D ABs, the interaction was significant (p = 0.01); class C and D ABs were used more in the weaning (p = 0.07) than in the fattening phase (p = 0.003). For the weaning phase, the use of class C and D ABs was greater in the PRRS+ than in the PRRS– chain (p < 0.01). In conclusion, PRRS status affected the growth of pigs and economic performance. Moreover, PRRS status significantly influenced the use of ABs during all the growing periods with the greatest impact being on the weaning phase

Investigation of the Defatted Colostrum 1H-NMR Metabolomics Profile of Gilts and Multiparous Sows and Its Relationship with Litter Performance

The aim of the study was to characterize the soluble metabolomics profile of defatted colostrum of sows at different parity number (PA) and to correlate the metabolomics profile with the Brix percentage estimate of colostrum immunoglobulin G (IgG) and sow productive traits. A total of 96 Meidam (crossbreed Large White 7 Meishan) sows of PA from 1-4 (PA1: 28; PA2:26; PA3:12; PA4:26) were included, and their productive traits were recorded at 10 days post-farrowing. Colostrum IgG was quantified using a Brix refractometer, and metabolomics profile was assessed using 1H-NMR spectroscopy. Sows' PA slightly influenced the metabolomics profile of colostrum. lactose and glycine were higher in PA1 compared with PA4 (p 0.05) and N-acetylglucosamine (GlcNAc) tended to be higher in PA2 than PA3 and PA4 (p < 0.10). The Brix percentage of IgG was negatively associated with lactose and positively with creatine, myo-inositol, and O-phosphocholine (p < 0.05). Taurine was positively related to litter weight at birth. GlcNAc and myo-inositol were linked to piglet mortality at day 10 with a negative and positive trend, respectively. In conclusion, colostrum of gilts and multiparous sows had a similar metabolomics profile. Specific metabolites contributed to explanation of the variability in colostrum Brix percentage estimate of IgG concentration and the sows' productive performance

Faecal Microbiota Characterisation of Potamochoerus porcus Living in a Controlled Environment

Intestinal bacteria establish a specific relationship with the host animal, which causes the acquisition of gut microbiota with a unique composition classified as the enterotype. As the name suggests, the Red River Hog is a wild member of the pig family living in Africa, in particular through the West and Central African rainforest. To date, very few studies have analysed the gut microbiota of Red River Hogs (RRHs) both housed under controlled conditions and in wild habitats. This study analysed the intestinal microbiota and the distribution of Bifidobacterium species in five Red River Hog (RRH) individuals (four adults and one juvenile), hosted in two different modern zoological gardens (Parco Natura Viva, Verona, and Bioparco, Rome) with the aim of disentangling the possible effects of captive different lifestyle and host genetics. Faecal samples were collected and studied both for bifidobacterial counts and isolation by means of culture-dependent method and for total microbiota analysis through the high-quality sequences of the V3-V4 region of bacterial 16S rRNA. Results showed a host-specific bifidobacterial species distribution. Indeed, B. boum and B. thermoacidophilum were found only in Verona RRHs, whereas B. porcinum species were isolated only in Rome RRHs. These bifidobacterial species are also typical of pigs. Bifidobacterial counts were about 106 CFU/g in faecal samples of all the individuals, with the only exception for the juvenile subject, showing 107 CFU/g. As in human beings, in RRHs a higher count of bifidobacteria was also found in the young subject compared with adults. Furthermore, the microbiota of RRHs showed qualitative differences. Indeed, Firmicutes was found to be the dominant phylum in Verona RRHs whereas Bacteroidetes was the most represented in Roma RRHs. At order level, Oscillospirales and Spirochaetales were the most represented in Verona RRHs compared with Rome RRHs, where Bacteroidales dominated over the other taxa. Finally, at the family level, RRHs from the two sites showed the presence of the same families, but with different levels of abundance. Our results highlight that the intestinal microbiota seems to reflect the lifestyle (i.e., the diet), whereas age and host genetics are the driving factors for the bifidobacterial population

Disentangling the Possible Drivers of Indri indri Microbiome: A Threatened Lemur Species of Madagascar

Research on the gut microbiome may help with increasing our understanding of primate health with species’ ecology, evolution, and behavior. In particular, microbiome-related information has the potential to clarify ecology issues, providing knowledge in support of wild primates conservation and their associated habitats. Indri (Indri indri) is the largest extant living lemur of Madagascar. This species is classified as “critically endangered” by the IUCN Red List of Threatened Species, representing one of the world’s 25 most endangered primates. Indris diet is mainly folivorous, but these primates frequently and voluntarily engage in geophagy. Indris have never been successfully bred under human care, suggesting that some behavioral and/or ecological factors are still not considered from the ex situ conservation protocols. Here, we explored gut microbiome composition of 18 indris belonging to 5 different family groups. The most represented phyla were Proteobacteria 40.1 ± 9.5%, Bacteroidetes 28.7 ± 2.8%, Synergistetes 16.7 ± 4.5%, and Firmicutes 11.1 ± 1.9%. Further, our results revealed that bacterial alpha and beta diversity were influenced by indri family group and sex. In addition, we investigated the chemical composition of geophagic soil to explore the possible ecological value of soil as a nutrient supply. The quite acidic pH and high levels of secondary oxide-hydroxides of the soils could play a role in the folivorous diet’s gut detoxification activity. In addition, the high contents of iron and manganese found the soils could act as micronutrients in the indris’ diet. Nevertheless, the concentration of a few elements (i.e., calcium, sulfur, boron, nickel, sodium, and chromium) was higher in non-geophagic than in geophagic soils. In conclusion, the data presented herein provide a baseline for outlining some possible drivers responsible for the gut microbiome diversity in indris, thus laying the foundations for developing further strategies involved in indris’ conservation

SARS-CoV-2 infects human cardiomyocytes promoted by inflammation and oxidative stress

INTRODUCTION The respiratory illness triggered by severe acute respiratory syndrome virus-2 (SARS-CoV-2) is often particularly serious or fatal amongst patients with pre-existing heart conditions. Although the mechanisms underlying SARS-CoV-2-related cardiac damage remain elusive, inflammation (i.e. 'cytokine storm') and oxidative stress are likely involved. METHODS AND RESULTS Here we sought to determine: 1) if cardiomyocytes are targeted by SARS-CoV-2 and 2) how inflammation and oxidative stress promote the viral entry into cardiac cells. We analysed pro-inflammatory and oxidative stress and its impact on virus entry and virus-associated cardiac damage from SARS-CoV-2 infected patients and compared it to left ventricular myocardial tissues obtained from non-infected transplanted hearts either from end stage heart failure or non-failing hearts (donor group). We found that neuropilin-1 potentiates SARS-CoV-2 entry into human cardiomyocytes, a phenomenon driven by inflammatory and oxidant signals. These changes accounted for increased proteases activity and apoptotic markers thus leading to cell damage and apoptosis. CONCLUSION This study provides new insights into the mechanisms of SARS-CoV-2 entry into the heart and defines promising targets for antiviral interventions for COVID-19 patients with pre-existing heart conditions or patients with co-morbidities

Epigenome-wide meta-analysis of blood DNA methylation and its association with subcortical volumes:findings from the ENIGMA Epigenetics Working Group

DNA methylation, which is modulated by both genetic factors and environmental exposures, may offer a unique opportunity to discover novel biomarkers of disease-related brain phenotypes, even when measured in other tissues than brain, such as blood. A few studies of small sample sizes have revealed associations between blood DNA methylation and neuropsychopathology, however, large-scale epigenome-wide association studies (EWAS) are needed to investigate the utility of DNA methylation profiling as a peripheral marker for the brain. Here, in an analysis of eleven international cohorts, totalling 3337 individuals, we report epigenome-wide meta-analyses of blood DNA methylation with volumes of the hippocampus, thalamus and nucleus accumbens (NAcc)-three subcortical regions selected for their associations with disease and heritability and volumetric variability. Analyses of individual CpGs revealed genome-wide significant associations with hippocampal volume at two loci. No significant associations were found for analyses of thalamus and nucleus accumbens volumes. Cluster-based analyses revealed additional differentially methylated regions (DMRs) associated with hippocampal volume. DNA methylation at these loci affected expression of proximal genes involved in learning and memory, stem cell maintenance and differentiation, fatty acid metabolism and type-2 diabetes. These DNA methylation marks, their interaction with genetic variants and their impact on gene expression offer new insights into the relationship between epigenetic variation and brain structure and may provide the basis for biomarker discovery in neurodegeneration and neuropsychiatric conditions