Pau Obregon-Gutierrez

Streptococcus suis is a Gram-positive bacterium with a dual role as a commensal member of the porcine nasal microbiota and a pathogen causing systemic disease in pigs and humans. Mounting evidence suggests that metabolism is a key driver of S. suis pathogenicity. Given the species' high genetic variability, we hypothesize that differences in metabolic networks could explain the diverse pathogenic phenotypes observed across different strains. To test this, we generated an atlas of over 3000 strain-specific and automatically curated genome-scale metabolic models that cover the breadth of pathogenic and commensal S. suis lineages. Using this model atlas, we performed the first species-level examination of metabolic traits in S. suis. Our simulations, supported by experimental validation, revealed three key insights. First, while metabolic traits are broadly conserved in S. suis, there are nevertheless lineage-dependent differences in amino acid auxotrophies and carbon utilization patterns that point towards distinct in vivo niches. Second, most strains are predicted to grow in different plausible in vivo environments regardless of their virulence phenotype, suggesting that metabolism is a weak barrier to systemic infection. Third, by systematically predicting reaction essentiality in more than 15 million reaction-strain-condition combinations, we identify a subset of 17 reactions, largely in nucleotide metabolism, that are conditionally essential in vivo and may serve as new targets for the development of new antimicrobials or vaccines. Overall, this study provides a valuable new resource for broadly examining S. suis metabolism and its role in pathogenicity.

Antimicrobial resistance (AMR) is a major public health concern influenced by antimicrobial use (AMU) in animal production systems. In swine, metaphylactic treatments may contribute to the emergence and dissemination of resistance genes. In this study, we isolated bacteria from the nasal cavities of 50 sows across 10 farms in the Federal District, Brazil. A total of 132 bacterial isolates were obtained and tested for phenotypic resistance to 23 antimicrobials using the disk diffusion method. Resistance was detected against all tested antimicrobials, with an overall resistance rate of 55.6% (1605/2888 tests). The highest resistance rates were observed for bacitracin (92.4%) and penicillin (79.2%), while lower resistance rates were found for aminoglycosides. Most isolates exhibited multidrug resistance to 7–9 classes of antimicrobials, including strains of Staphylococcus, Escherichia coli, and Klebsiella—all of which are relevant in the context of One Health. Actinobacillus suis showed the highest resistance levels among all identified species. AMR was positively correlated with both the duration and the number of antimicrobial agents used in feed, reinforcing the need for prudent AMU practices. The use of autogenous vaccines against Pasteurella multocida was associated with reduced lung lesions, underscoring the value of vaccination in disease control. AMR surveillance programs may benefit from including bacterial colonizers from the microbiota, though further studies are necessary to better understand the resistance dynamics of these commensals.

Antimicrobial resistance is a universal threat and is leading to a new awareness of antimicrobial use. The colonization of tissues by some microorganisms carrying resistance genes may pose a risk of spreading resistance to pathogens. Antimicrobials may induce an unstable microbiome that compromises the animal's immunity. Indeed, dysbiosis has been linked to many alterations in the immune response. Here, we isolated bacterial colonizers from the nasal microbiota of sows to describe the phenotypic resistance profile on different health managements. One hundred and thirty-two strains isolated from 50 nasal swabs collected from sows were tested against up to 23 antimicrobial agents by disk diffusion. Overall, the nasal communities showed 55% antimicrobial resistance (1605/2888 tests). Resistance was detected for all tested antimicrobials. The antimicrobial showing higher rate of resistance was bacitracin (92%), while the lowest was found in the aminoglycosides. Actinobacillus suis was one of the species with highest rate of resistance. The time and number of drugs used in feed influenced the resistance rate in the isolates. Vaccination was confirmed as a strategic protocol for disease control Studies on antimicrobial resistance in commensals may allow the identification of microorganisms to be used in surveillance and may constitute a tool for evaluating strategies to reduce antimicrobial use in pig herds.

Mounting evidence suggests that metabolite exchange between microbiota members is a key driver of microbiota composition. However, we still know little about the metabolic interaction networks that occur within many microbiota. This is particularly true for the nasal microbiota, and current efforts towards this end are hampered by a lack of microbial consortia that would enable the mapping of metabolic interactions between nasal microbiota members under in vitro conditions. To tackle these issues, we developed the Porcine Nasal Consortium (PNC8), a rationally designed microbial consortium of eight strains representing the most in vivo abundant genera in the nasal microbiota of healthy piglets. We used this consortium to systematically examine the metabolic capabilities of nasal microbiota members, as well as the metabolic interactions occurring between them. We found that PNC8 strains differ substantially in their metabolic pathway repertoire and ability to grow across various in vitro conditions. Nevertheless, spent-media experiments revealed that most metabolic interactions between PNC8 strains are negative, and exometabolomics data pointed to co-depletion of sugars as a key driver of this interaction network. Finally, direct co-cultivation experiments showed that, as a result of this largely negative metabolic interaction network, competition is common among pairs of PNC8 strains and leads to a complex competition hierarchy in which only few strains are able to consistently outcompete all others. Overall, this work provides a valuable resource for studying the nasal microbiota under experimentally tractable in vitro conditions and is a key step towards mapping its metabolic interaction network.

The nasal microbiota is a key contributor to animal health, and characterizing the nasal microbiota composition is an important step towards elucidating the role of its different members. Efforts to characterize the nasal microbiota composition of domestic pigs and other farm animals frequently report the presence of bacteria that are typically found in the gut, including many anaerobes from the Bacteroidales and Clostridiales orders. However, the in vivo role of these gut-microbiota associated taxa is currently unclear. Here, we tackled this issue by examining the prevalence, origin, and activity of these taxa in the nasal microbiota of piglets. First, analysis of the nasal microbiota of farm piglets sampled in this study, as well as various publicly available data sets, revealed that gut-microbiota associated taxa indeed constitute a substantial fraction of the pig nasal microbiota that is highly variable across individual animals. Second, comparison of herd-matched nasal and rectal samples at amplicon sequencing variant (ASV) level showed that these taxa are largely shared in the nasal and rectal microbiota, suggesting a common origin driven presumably by the transfer of fecal matter. Third, surgical sampling of the inner nasal tract showed that gut-microbiota associated taxa are found throughout the nasal cavity, indicating that these taxa do not stem from contaminations introduced during sampling with conventional nasal swabs. Finally, analysis of cDNA from the 16S rRNA gene in these nasal samples indicated that gut-microbiota associated taxa are indeed active in the pig nasal cavity. This study shows that gut-microbiota associated taxa are not only present, but also active, in the nasal cavity of domestic pigs, and paves the way for future efforts to elucidate the function of these taxa within the nasal microbiota.

Background: The nasal microbiota is a key contributor to animal health, and characterizing the nasal microbiota composition is an important step towards elucidating the role of its different members. Efforts to characterize the nasal microbiota composition of domestic pigs and other farm animals frequently report the presence of bacteria that are typically found in the gut, including many anaerobes from the Bacteroidales and Clostridiales orders. However, the in vivo role of these gut-microbiota associated taxa is currently unclear. Here, we tackled this issue by examining the prevalence, origin, and activity of these taxa in the nasal microbiota of piglets. Results: First, analysis of the nasal microbiota of farm piglets sampled in this study, as well as various publicly available data sets, revealed that gut-microbiota associated taxa indeed constitute a substantial fraction of the pig nasal microbiota that is highly variable across individual animals. Second, comparison of herd-matched nasal and rectal samples at amplicon sequencing variant (ASV) level showed that these taxa are largely shared in the nasal and rectal microbiota, suggesting a common origin driven presumably by the transfer of fecal matter. Third, surgical sampling of the inner nasal tract showed that gut-microbiota associated taxa are found throughout the nasal cavity, indicating that these taxa do not stem from contaminations introduced during sampling with conventional nasal swabs. Finally, analysis of cDNA from the 16S rRNA gene in these nasal samples indicated that gut-microbiota associated taxa are indeed active in the pig nasal cavity. Conclusion: This study shows that gut-microbiota associated taxa are not only present, but also active, in the nasal cavity of domestic pigs, and paves the way for future efforts to elucidate the in vivo function of these taxa within the nasal microbiota.

The microbiota plays an important role in the development of diarrhea in pre-weaned calves. The characterization of the fecal microbiota in health and disease can be critical to unravel the bacterial dynamics associated with diarrhea and help with its prevention and control. In this study, we aimed to detect changes in the fecal microbiota of calves that experienced early-life diarrhea episodes. Fecal samples were taken from calves remaining healthy and calves with an episode of diarrhea during the study. We sampled at arrival (12 days of age) and after one and two months of life; also, at the time of the diarrhea episode for the diarrheic calves (day 17). Samples were processed to extract total DNA, submitted to 16S rRNA gene sequencing, and bioinformatically analyzed to infer the bacterial populations. Microbiota changes through time were reported for both groups. However, we detected an earlier stabilization in the healthy group. Moreover, we detected changes within low abundant taxa that may play a role in the subsequent health status of the animals. The fecal microbiota of healthy and diarrheic calves showed different dynamics in the diversity through time that may be the reflections of the variations within low-abundant taxa.

The nasal microbiota composition is associated with the health status of piglets. Sow-contact in early life is one of the factors influencing the microbial composition in piglets; however, its impact has never been assessed in the nasal microbiota of piglets reared in controlled environmental conditions. Nasal microbiota of weaning piglets in high-biosecurity facilities with different time of contact with their sows (no contact after farrowing, contact limited to few hours or normal contact until weaning at three weeks) was unveiled by 16S rRNA gene sequencing. Contact with sows demonstrated to be a major factor affecting the nasal microbial composition of the piglets. The nasal microbiota of piglets that had contact with sows until weaning, but were reared in high biosecurity facilities, was richer and more similar to the previously described healthy nasal microbiota from conventional farm piglets. On the other hand, the nasal communities inhabiting piglets with no or limited contact with sows was different and dominated by bacteria not commonly abundant in this body site. Furthermore, the length of sow–piglet contact was also an important variable. In addition, the piglets raised in BSL3 conditions showed an increased richness of low-abundant species in the nasal microbiota. Artificially rearing in high biosecurity facilities without the contact of sows as a source of nasal colonizers had dramatic impacts on the nasal microbiota of weaning piglets and may introduce significant bias into animal research under these conditions.