Enterococcus Cecorum Research Articles

The gut microbiota of wild birds undergoing rehabilitation as a reservoir of multidrug-resistant enterococci in a metropolitan area in Brazil.

Enterococci are ubiquitous usually commensal bacteria that can act as opportunistic pathogens frequently associated with resistance to multiple antimicrobial classes. A variety of animals may carry potentially harmful enterococci. In the present work, the occurrence and characteristics of enterococci recovered from the fecal microbiota of wild birds belonging to four families (Accipitridae, Cathartidae, Falconidae and Strigidae) were investigated. Enterococci were recovered from 104 (92.0%) fecal samples obtained from 113 birds, and 260 strains were selected for additional characterization. Enterococcus faecalis was the predominant species (63.8%), followed by Enterococcus hirae (16.2%), Enterococcus faecium (11.5%), Enterococcus gallinarum (5.4%), Enterococcus avium (1.5%), Enterococcus casseliflavus (0.8%), and Enterococcus raffinosus and Enterococcus cecorum (0.4% each). Major percentages (11.9% 75.0%) of nonsusceptibility were observed to quinolones (particularly to enrofloxacin), erythromycin, rifampin, nitrofurantoin, tetracycline and streptomycin. Gentamicin and ampicillin resistances (13.3% each) were only detected among E. faecium. A total of 133 (51.2%) strains were MDR, showing a large variety of MDR profiles, composed by simultaneous resistance encompassing 3 to 12 antimicrobials. MDR strains were found in 68.2% of the birds. Antimicrobial resistance was associated with the presence of the aac(6')-aph(2″)-Ia, aph(2″)-Id, ant(6)-Ia, ant(9)-Ia, ant(9)-Ib, tet(M), tet(L), tet(S), erm(B), mef(A/E), msrC, and vat(D) genes. The most common virulence genes were efaA, gelE, ace, eeP, and asa1. PFGE analysis revealed a large genetic diversity among most of the strains. MLST performed for 35 E.faecalis strains revealed 23 different STs, whereas 14 STs were found among 18 E.faecium strains. Hospital-associated lineages ST22, ST25, ST56, ST1274 were identified. The results show that the wild birds investigated can carry a diversity of potentially hazardous enterococcal strains displaying multiple antimicrobial resistance and virulence genes, reinforcing the assumption that these animals provide an important target to monitor the circulation of microorganisms that deserve consideration under the One Health perspective.

Elevated muramyl dipeptide by sialic acid-facilitated postantibiotic pathobiont expansion contributes to gut dysbiosis-induced mastitis in mice

IntroductionIn responses to antibiotics exposure, gut dysbiosis is a risk factor not only for pathogen infection but also for facilitating pathobiont expansion, resulting in increased inflammatory responses in the gut and distant organs. However, how this process is regulated has not been fully elucidated. ObjectivesIn this study, we investigated the role of sialic acid, a host-derived carbohydrate, in the pathogenesis of gut dysbiosis-derived inflammation in distant organs. MethodsAmpicillin (Amp)-induced gut dysbiotic mice were treated with N-glycolylneuraminic acid (Neu5Gc) and N-acetylneuraminic acid (Neu5Ac) for three weeks to assess the role of sialic acids in mastitis. The underlying mechanism by which sialic acids regulate mastitis was explored using 16S rRNA sequencing, transcriptomics and employed multiple molecular approaches. ResultsAdministration of Neu5Ac and Neu5Gc exacerbated gut dysbiosis-induced mastitis and systemic inflammation. The gut dysbiosis caused by Amp was also aggravated by sialic acid. Notably, increased Enterococcus expansion, which was positively correlated with inflammatory markers, was observed in both Neu5Ac- and Neu5Gc-treated gut dysbiotic mice. Treatment of mice with Enterococcus cecorum (E. cecorum) aggravated gut dysbiosis-induced mastitis. Mechanically, sialic acid-facilitated E. cecorum expansion promoted muramyl dipeptide (MDP) release, which induced inflammatory responses by activating the NOD2-RIP2-NF-κB axis. ConclusionsCollectively, our data reveal a role of sialic acid-facilitated postantibiotic pathobiont expansion in gut dysbiosis-associated inflammation, highlighting a potential strategy for disease prevention by regulating the MDP-NOD2-RIP2 axis.

Research Note: Comparison of Enterococcus cecorum genomes from broiler chickens with enterococcal spondylitis in Australian farms and strains from other countries

Chickens in Australia have recently been identified with symptoms and morphological findings including spondylitis attributed to pathogenic Enterococcus cecorum. Notably, there is limited information on clinical E. cecorum strains in Australia. The cpsO gene, located downstream of the capsular polysaccharide (cps) locus, was recently reported to successfully differentiate between pathogenic and commensal E. cecorum strains, as this gene is highly conserved in the pathogenic strains. In this study, pathogenic E. cecorum, with a conserved cpsO gene, was detected on 1 of the 2 farms studied in Australia. E. cecorum strains isolated from clinical sites of the diseased birds from the second farm did not have the cpsO gene and were distant from the isolates of the first farm. A cpsO PCR of the caecal content of the birds on this farm was positive, while cpsO PCR of washed culture plates where the tissue extracts were spread onto and incubated for bacterial growth was negative. This suggests that pathogenic E. cecorum with the cpsO gene, as detected in Farm 1 and reported in other countries, was present in the second farm but could not grow on the selective agar plates during the initial step of E. cecorum isolation. Nevertheless, E. cecorum isolated from the clinical sites on the second farm might represent the pathogenic strain, but further animal studies are required to validate this possibility. Phylogenetic analysis showed that the pathogenic strains in Australia were most closely related to the clinical strains in North America.

Towards the identification of transmission pathways and early detection of Enterococcus cecorum infection in broiler chickens

Enterococcus cecorum (EC) infection is an emerging endemic disease in UK and global broiler poultry with major economic impact and welfare concerns. There are significant research gaps with regards to EC pathogenesis, source of infection, transmission routes and early detection of disease, which this study aimed to address. In this prospective study, 725 environmental samples were collected from 4 broiler farms (A-D) the day before chick placement (d 1) and through the subsequent crop (d 7, 14, and 21). Cecal swabs were collected from birds that died of natural causes during the study period. A sample of birds that had been found dead or were culled for health reasons, were presented for post-mortem and samples were taken from lesions for EC culture. DNA was extracted from all environmental samples and EC detected using a qPCR and MALDI-TOF. Two EC isolates from diseased birds were inoculated on concrete slabs and incubated at 23°C and 32°C followed by swabbing of concrete culturing and determination of EC cfu at defined time points. Alongside environmental and bird sampling commercially available, smart camera systems were installed in selected houses on each farm to monitor bird activity and distribution. No EC outbreak occurred during the study, however, it was detected by qPCR in 215/725 (29.7 %) of all samples collected. Also, EC DNA was detected on average in 37% of samples collected on d 1, with approx. 88% of samples from chick paper being positive. Despite this, it was only cultured from 3 ceca samples and joint fluids of two infected birds from farm B on d 14 and 21. The survival experiments using isolates from infected chickens showed EC can survive on concrete for at least 21 d. This study provides invaluable insights into transmission pathways and tenacity of EC. Further studies are needed to determine strain characteristics in relation to their ability to cause disease and to further elucidate the sources of infection on poultry farms.

A detailed analysis of 16S rRNA gene sequencing and conventional PCR-based testing for the diagnosis of bacterial pathogens and discovery of novel bacteria.

This study represents the first analysis of the bacterial community in chickens affected by swollen head syndrome, utilizing 16S rRNA gene sequencing. Samples were obtained from clinical laying chickens and were examined for the presence of Avibacterium paragallinarum (APG) and Ornithobacterium rhinotracheale (ORT) using conventional polymerase chain reaction (PCR). From the samples, five APG-positive (APG) and APG-negative (N-APG) samples were chosen, along with five specific pathogen-free chickens, for 16S rRNA gene sequencing. Results showed that APG and ORT were widely detected in the chicken samples with swollen head syndrome (SHS, 9/10), while APG was detected in all five specific pathogen-free (SPF) samples. In contrast, conventional PCR sensitivity was found to be inadequate for diagnosis, with only 35.7% (5/14) and 11.1% (1/9) sensitivity for APG and ORT, respectively, based on 16S rRNA gene sequencing data. Furthermore, 16S rRNA gene sequencing was able to quantify the bacteria in the samples, revealing that the relative abundance of APG in the APG group ranged from 2.7 to 81.3%, while the relative abundance of APG in the N-APG group ranged from 0.1 to 21.0%. Notably, a low level of APG was also detected in all 5 SPF samples. The study also identified a significant number of animal and human common bacterial pathogens, including but not limited to Gallibacterium anatis, Riemerella columbina, Enterococcus cecorum, Mycoplasma synoviae, Helicobacter hepaticus, and Staphylococcus lentus. In conclusion, 16S rRNA gene sequencing is a valuable tool for bacterial pathogen diagnosis and the discovery of novel bacterial pathogens, while conventional PCR is not reliable for diagnosis.

51. Synthesis: Enterococcus cecorum, an opportunistic pathogen of poultry: Learning more about to better control it on the farms

51. Synthesis: Enterococcus cecorum, an opportunistic pathogen of poultry: Learning more about to better control it on the farms

91. Preliminary study of the molecular diversity of Enterococcus cecorum in Brittany farms

91. Preliminary study of the molecular diversity of Enterococcus cecorum in Brittany farms

82. Effect of a triple-strain Bacillus-based probiotic on in vitro inhibition of clinical Enterococcus cecorum strains isolated from commercial broilers

82. Effect of a triple-strain Bacillus-based probiotic on in vitro inhibition of clinical Enterococcus cecorum strains isolated from commercial broilers

Aspirin prevents colorectal cancer by regulating the abundance of Enterococcus cecorum and TIGIT+Treg cells

Although aspirin can reduce the incidence of colorectal cancer (CRC), there is still uncertainty about its significance as a treatment for CRC, and the mechanism of aspirin in CRC is not well understood. In this study, we used aspirin to prevent AOM/DSS-induced CRC in mice, and the anti-CRC efficacy of aspirin was assessed using haematoxylin and eosin (H&E) staining and by determining the mouse survival rate and tumour size. 16S rDNA sequencing, flow cytometry (FCM), and Western blotting were also conducted to investigate the changes in the gut microbiota, tumour immune microenvironment, and apoptotic proteins, respectively. The results demonstrated that aspirin significantly exerted anti-CRC effects in mice. According to 16S rDNA sequencing, aspirin regulated the composition of the gut microbiota and dramatically reduced the abundance of Enterococcus cecorum. FCM demonstrated that there were more CD155 tumour cells and CD4 + CD25 + Treg cells showed increased TIGIT levels. Moreover, increased TIGIT expression on Treg cells is associated with reduced Treg cell functionality. Importantly, the inhibition of Treg cells is accompanied by the promotion of CD19 + GL-7 + B cells, CD8 + T cells, CD4 + CCR4 + Th2 cells, and CD4 + CCR6 + Th17 cells. Overall, aspirin prevents colorectal cancer by regulating the abundance of Enterococcus cecorum and TIGIT + Treg cells.

The Genetic Landscape of Antimicrobial Resistance Genes in Enterococcus cecorum Broiler Isolates.

Enterococcus cecorum is associated with bacterial chondronecrosis with osteomyelitis (BCO) in broilers. Prophylactic treatment with antimicrobials is common in the poultry industry, and, in the case of outbreaks, antimicrobial treatment is needed. In this study, the minimum inhibitory concentrations (MICs) and epidemiological cutoff (ECOFF) values (COWT) for ten antimicrobials were determined in a collection of E. cecorum strains. Whole-genome sequencing data were analyzed for a selection of these E. cecorum strains to identify resistance determinants involved in the observed phenotypes. Wild-type and non-wild-type isolates were observed for the investigated antimicrobial agents. Several antimicrobial resistance genes (ARGs) were detected in the isolates, linking phenotypes with genotypes for the resistance to vancomycin, tetracycline, lincomycin, spectinomycin, and tylosin. These detected resistance genes were located on mobile genetic elements (MGEs). Point mutations were found in isolates with a non-wild-type phenotype for enrofloxacin and ampicillin/ceftiofur. Isolates showing non-wild-type phenotypes for enrofloxacin had point mutations within the GyrA, GyrB, and ParC proteins, while five amino acid changes in penicillin-binding proteins (PBP2x superfamily) were observed in non-wild-type phenotypes for the tested β-lactam antimicrobials. This study is one of the first that describes the genetic landscape of ARGs within MGEs in E. cecorum, in association with phenotypical resistance determination.

In Vitro Evaluation of Phytobiotic Mixture Antibacterial Potential against Enterococcus spp. Strains Isolated from Broiler Chicken.

Enterococcus spp. are normal intestinal tract microflorae found in poultry. However, the last decades have shown that several species, e.g., Enterococcus cecorum, have become emerging pathogens in broilers and may cause numerous losses in flocks. In this study, two combinations (H1 and H2) of menthol, 1,8-cineol, linalool, methyl salicylate, γ-terpinene, p-cymene, trans-anethole, terpinen-4-ol and thymol were used in an in vitro model, analyzing its effectiveness against the strains E. cecorum, E. faecalis, E. faecium, E. hirae and E. gallinarum isolated from broiler chickens from industrial farms. To identify the isolated strains classical microbiological methods and VITEK 2 GP cards were used. Moreover for E. cecorum a PCR test was used.. Antibiotic sensitivity (MIC) tests were performed for all the strains. For the composition H1, the effective dilution for E. cecorum and E. hirae strains was 1:512, and for E. faecalis, E. faecium and E. gallinarum, 1:1024. The second mixture (H2) showed very similar results with an effectiveness at 1:512 for E. cecorum and E. hirae and 1:1024 for E. faecalis, E. faecium and E. gallinarum. The presented results suggest that the proposed composition is effective against selected strains of Enterococcus in an in vitro model, and its effect is comparable to classical antibiotics used to treat this pathogen in poultry. This may suggest that this product may also be effective in vivo and provide effective support in the management of enterococcosis in broiler chickens.

Detection of Enterococcus cecorum to identify persistently contaminated locations using faecal and environmental samples in broiler houses of clinically healthy flocks

ABSTRACT Worldwide outbreaks make infections with pathogenic strains of Enterococcus cecorum (EC) one of the most important diseases in the broiler industry. Although research has increased knowledge about the pathogen, the transmission is not fully understood. Samples from different locations were collected from two broiler farms in Germany over a total of six production cycles. Samples were collected at days 1, 5, 10, 15, 21, 27, 34, 41 post-hatch and after cleaning and disinfection (C&D). A total of 1017 samples were collected from 25 different locations on the farms. Samples were analysed in the laboratory for EC by quantitative real-time PCR. Overall, 7.5% of the samples were positive. The probabilities for positive and negative samples did not differ between the farms. The number of findings differed significantly between the cycles. Compared to other samples, the chances of detecting EC in faecal samples were significantly higher. Most positive samples were found in the last week of the production periods, indicating an accumulation of EC in the barn environment. After C&D, positive PCR results were obtained in four out of 14 locations. A re-introduction from contaminated environment seemed possible. However, one pooled faecal sample was positive 1 day post-hatch. The locations that showed positive results after C&D and the positive faecal sample 1 day post-hatch indicated the persistence of EC in broiler houses of clinically healthy flocks that could lead to potential horizontal transmission routes. The present study detected potential EC sources and may help to improve hygienic measures to avoid transmissions. RESEARCH HIGHLIGHTS Methodology is suitable to detect EC during production and after C&D. Locations were detected that may serve as a reservoir for EC. Cycles with fewer positive samples were observed. Cleaning and disinfection had a major impact on the detection of EC.

Enterococcus Cecorum bacteremia and meningitis in an asplenic patient: A case report

This paper presents a case of Enterococcus cecorum (E. cecorum) meningitis with bacteremia in a middle-aged woman who had previously undergone a splenectomy. Within 24 hours, the bacterium was identified from a blood culture. The patient was clinically diagnosed with meningitis, which was radiologically verified by an MRI brain scan. Fortunately, she had a favorable outcome. To the best of our knowledge, this is the first reported case of E. cecorum-induced meningitis in humans and the second documented incidence of E. cecorum infection in humans in Australia. The purpose of this case report is to emphasize the potential for serious human infections caused by an extremely rare pathogen.

Molecular Genomic Analyses of Enterococcus cecorum from Sepsis Outbreaks in Broilers.

Extensive genomic analyses of Enterococcus cecorum isolates from sepsis outbreaks in broilers suggest a polyphyletic origin, likely arising from core genome mutations rather than gene acquisition. This species is a normal intestinal flora of avian species with particular isolates associated with osteomyelitis. More recently, this species has been associated with sepsis outbreaks affecting broilers during the first 3 weeks post-hatch. Understanding the genetic and management basis of this new phenotype is critical for developing strategies to mitigate this emerging problem. Phylogenomic analyses of 227 genomes suggest that sepsis isolates are polyphyletic and closely related to both commensal and osteomyelitis isolate genomes. Pangenome analyses detect no gene acquisitions that distinguish all the sepsis isolates. Core genome single nucleotide polymorphism analyses have identified a number of mutations, affecting the protein-coding sequences, that are enriched in sepsis isolates. The analysis of the protein substitutions supports the mutational origins of sepsis isolates.

Serological monitoring of Enterococcus cecorum specific antibodies in chickens

Pathogenic Enterococcus cecorum (EC) has gained increasing importance as the cause of skeletal infections in meat-type chicken production. Since effective intervention strategies are scarce, it must be focused on preventive measures. Vaccination of meat-type breeder chicken flocks is common practice to protect the progeny against infection with EC. However, no data are available on seroconversion after infection or vaccination. The aim of the present study was the serological monitoring of chickens for EC-specific immunoglobulin Y (IgY) using a newly established EC-specific, indirect ELISA for chickens. Sera from previous infection studies were used for the establishment of the assay. Serum samples from confirmed EC-positive meat-type chicken flocks, vaccinated, and non-vaccinated meat-type chicken breeder flocks were analyzed for EC-specific IgY. Comparison of ELISA results with results from real-time PCR and/or bacteriological examination via culture revealed fair to substantial agreement. In infected chickens, more samples were classified as positive via ELISA than via real-time PCR and/or bacteriological examination via culture. Focusing on chickens experimentally infected at 1 day post-hatch (dph), the highest proportion of positive results and highest S/P ratios were found at 42 dph (p < 0.05). A similar trend was observed for the samples from naturally infected chickens (p < 0.05). Adjustment of the secondary antibody against immunoglobulin M (IgM) may open possibilities to use the assay during the early phase of the growing period, when there is still a chance to treat the infection. The examination of samples from vaccinated and non-vaccinated meat-type breeder chickens revealed no significant differences of S/P ratios independent of farm and autogenous vaccine used. In addition to that, monitoring of a non-vaccinated meat-type breeder chicken flock at 4, 10, 15, and 19 weeks post-hatch showed a continuous increase of ELISA-positive serum samples associated with an increase of S/P ratios. This may be explained by cross reactivity with antibodies to Enterococcus hirae or natural antibodies. The usage of EC-specific, recombinant proteins for coating of the plates may help to reduce unspecific background and increase the assay’s specificity in future applications. In conclusion, the newly developed ELISA provides a suitable tool for serological monitoring of meat-type chickens during experimental studies with EC under standardized conditions. Remarkably, the assay is able to detect a higher proportion of EC-positive chickens than other methods, which are currently available. However, the assay is not yet suitable for the monitoring of breeder flocks due to high background.

Enterococcus cecorum lesion strains are less sensitive to the hostile environment of albumen and more resistant to lysozyme compared to cloaca strains

ABSTRACT Enterococcus cecorum lesion strains cause more embryonic mortality after inoculation into the albumen of embryonated eggs compared to cloaca strains. We hypothesized that these strain differences are a result of differences in sensitivity to the antimicrobial effects of the albumen. In this study, the sensitivity of 14 lesion strains and 14 cloaca strains to albumen from 12-day incubated and non-incubated eggs was assessed. A major antimicrobial protein of the albumen is lysozyme and, therefore, the lysozyme resistance of all strains was determined. Albumen from 12-day incubated and non-incubated eggs was inoculated with four cloaca strains and four lesion strains (104 CFU/tube). Based on the results, in a subsequent experiment, all 28 strains were inoculated only into albumen from non-incubated eggs. For all strains, the minimal inhibitory concentrations (MIC) of lysozyme were determined using an agar dilution method and the growth rates in broth with (500 and 2000 µg/ml) or without lysozyme were assessed. Compared to cloaca strains, lesion strains had 320 times higher odds of being reisolated from albumen (95% confidence interval: 55–3962) and had 8.5 times higher reisolation ratios (reisolation dose/inoculation dose) (95% confidence interval: 4.8–15.3). Thirteen cloaca strains had a MIC ranging from 1000 to 8000 µg/ml, while one cloaca strain and all lesion strains were resistant to the lysozyme concentrations tested. Growth rates of cloaca strains were decreased more by lysozyme compared to lesion strains. In conclusion, lesion strains had higher survival rates in egg albumen and were more resistant to lysozyme compared to cloaca strains. RESEARCH HIGHLIGHTS Egg albumen inhibits Enterococcus cecorum cloaca strains more than lesion strains. Enterococcus cecorum lesion strains are resistant to high concentrations of lysozyme. Lysozyme resistance could enhance survival in albumen and body fluids.

Research Note: Preliminary results, first detection of Enterococcus cecorum from environmental samples by streaking on X-Gluc containing selective media

The isolation of cultivable E. cecorum from the environment of poultry houses remains a challenge. Environmental samples (dust wipes, equipment swabs, pooled feces) and samples from culled bird vertebras were collected from an infected broiler flock on d 37 posthatching. To isolate the bacterium from the cultivable microbiota, suspensions from the environmental samples were streaked onto a blood agar base medium supplemented with 5-bromo-4-chloro-3-indolyl-beta-D-glucuronic acid cyclohexylammonium salt (X-Gluc), colistin sulfate, and nalidixin. The chromogenic reaction facilitated the isolation of E. cecorum from contaminated surfaces and pooled feces. Isolates from both the environment and vertebras were confirmed using MALDI-TOF and PCR analysis. Colony appearance and antimicrobial susceptibility tests revealed no phenotypic differences among the isolates. It remained unclear whether the isolates originated from the same clone. However, the principle of isolating the pathogen by streaking on a chromogenic agar may motivate researchers to investigate the transmission routes of infectious isolates, potentially leading to the optimization of biosecurity measures.

Vertebral osteomyelitis associated with Enterococcus faecalis in Broiler Breeders in Chile

Vertebral osteomyelitis is a re-emerging disease characterized by inflammation and necrosis of the thoracic vertebral body, caused by Enterococcus cecorum. Here, we report the first case of vertebral osteomyelitis caused by Enterococcus faecalis in Broiler Breeders, in Chile, which also causes infections in humans and is resistant to multiple antimicrobials, representing a risk to public health.

Specific Oral Microbial Differences in Proteobacteria and Bacteroidetes Are Associated with Distinct Sites When Moving from Healthy Mucosa to Oral Dysplasia-A Microbiome and Gene Profiling Study and Focused Review.

Oral potentially malignant disorders (OPMDs) are a group of conditions that carry a risk of oral squamous cell carcinoma (OSCC) development. Recent studies indicate that periodontal disease-associated pathogenic bacteria may play a role in the transition from healthy mucosa to dysplasia and to OSCC. Yet, the microbial signatures associated with the transition from healthy mucosa to dysplasia have not been established. To characterize oral microbial signatures at these different sites, we performed a 16S sequencing analysis of both oral swab and formalin-fixed, paraffin-embedded tissue (FFPE) samples. We collected oral swabs from healthy mucosa (from healthy patients), histologically normal mucosa adjacent to dysplasia, and low-grade oral dysplasia. Additionally, FFPE samples from histologically normal mucosa adjacent to OSCC, plus low grade and high-grade oral dysplasia samples were also collected. The collected data demonstrate significant differences in the alpha and beta microbial diversities of different sites in oral mucosa, dysplasia, and OSCC, as well as increased dissimilarities within these sites. We found that the Proteobacteria phyla abundance increased, concurrent with a progressive decrease in the Firmicutes phyla abundance, as well as altered levels of Enterococcus cecorum, Fusobacterium periodonticum, Prevotella melaninogenica, and Fusobacterium canifelinum when moving from healthy to diseased sites. Moreover, the swab sample analysis indicates that the oral microbiome may be altered in areas that are histologically normal, including in mucosa adjacent to dysplasia. Furthermore, trends in specific microbiome changes in oral swab samples preceded those in the tissues, signifying early detection opportunities for clinical diagnosis. In addition, we evaluated the gene expression profile of OSCC cells (HSC-3) infected with either P. gingivalis, T. denticola, F. nucelatum, or S. sanguinis and found that the three periodontopathogens enrich genetic processes related to cancer progression, including skin keratinization/cornification, while the commensal enriched processes related to RNA processing and adhesion. Finally, we reviewed the dysplasia microbiome literature and found a significant decrease in commensal bacteria, such as the Streptococci genus, and a simultaneous increase in pathogenic bacteria, mainly Bacteroidetes phyla and Fusobacterium genus. These findings suggest that features of the oral microbiome can serve as novel biomarkers for dysplasia and OSCC disease progression.

Characterizing the impact of Enterococcus cecorum infection during late embryogenesis on disease progression, cecal microbiome composition, and early performance in broiler chickens

Enterococcus cecorum (EC) has been associated with septicemia and early mortality in broiler chickens. There is limited research investigating the pathogenicity of EC field strains obtained from affected birds. The purpose of this study was to evaluate the effect of in-ovo administration into the amnion with different EC field isolates at d 18 of embryogenesis (DOE18). In Exp 1, 7 EC field isolates alone or in combination (EC1-EC3, EC4-EC5, EC6, and EC7) were selected based on phenotypic characteristics and evaluated at different concentrations (1×102, 1×104, and 1×106 CFU/200 µL/embryo) to assess the impact on early performance and macroscopic lesions. Three isolates (n=3; EC2, EC5, EC7) were selected for additional evaluation based on the significant (P < 0.05) BWG reduction (d 0-21) compared to the negative control (NC) and the presence of macroscopic lesions observed during posting sessions at d 14 and d 21. An additional isolate associated with enterococcal spondylitis was included in Exp 2 (EC11B). Treatment groups for Exp 2 include: 1) NC, 2) EC2, 3) EC5, 4) EC7, and 5) EC11B (n=90-120/embryos/group). Groups 2 to 5 were challenged at 1×102 CFU/200 µL/embryo by in-ovo injection into the amnion at DOE18. Chicks were placed in battery cages for the duration of the study (21 d), and pen weights were recorded at d 0, d 7, d 14, and d 21 to calculate average BW and BWG. At d 14 and d 21 posthatch, liver, spleen, free thoracic vertebrae (FTV), and femoral head (FH) were aseptically collected to enumerate Enterococcus spp. using Chromagar Orientation as the selective media. Cecal contents were collected at d 21 to evaluate the effect of EC challenge on the cecal microbiome composition. There was a significant (P < 0.05) reduction in BW at d 21, and BWG from d 14 to 21 and d 0 to 21, for EC7 and EC11B. Enterococcus cecorum was recovered from the FTV of all challenged groups at d 14 and d 21. The most representative lesions were pericarditis, hydropericardium, focal heart necrosis, and FH osteomyelitis. However, lesions were not uniform across challenged groups or ages (d 14 and d 21). Alpha diversity of the cecal contents was markedly lower in EC5 and EC11B compared to all treatment groups suggesting that EC exposure during late embryogenesis affect the cecal microbiome up to 21 d posthatch. Additionally, these results highlight the differences in pathogenicity of EC strains isolated from field cases and suggest that hatchery exposure to EC during late embryogenesis is a potential route of introduction into a flock.