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Characterization of Equine Platelet Lysate for Nebulization in the Horse

ABSTRACT: Platelet lysate (PL) demonstrates antimicrobial and anti-inflammatory properties and may offer potential for treatment of bacterial pneumonia in horses. It remains unknown whether nebulization represents a suitable method of administration of PL in horses. This study characterized particle size and flow rate of pooled equine PL nebulized using an equine-specific nebulizer (Flexivent). Protein composition and antimicrobial activity were compared before and after nebulization. Protein composition was evaluated according to growth factor, antimicrobial peptide and cytokine concentrations and proteomic analysis. To evaluate antimicrobial activity, bacterial growth inhibition [maximum growth (µmax); carrying capacity (K)] were determined for E.coli, Streptococcus equi subsp zooepidemicus and Rhodococcus equi (WT and MDR) using pre- and post-nebulized PL concentrations of 50%. Flow rate and median particle size were 0.8 ml/min and 4.991 µm with 52% of particles ≤ 5 µm. Cytokine, growth factor, and antimicrobial peptide concentrations and proteomic profiles were similar pre- and post-nebulization. For all bacteria, nebulization did not alter effect of PL on growth parameters. Negative effects of PL treatments on K were noted for S. zooepidemicus (p= 0.009 all) compared to BHI. No effect of PL was observed on µmax for S.zooepidemicus and on K and µmax for E.coli. For R. equi (WT and MDR), K and µ were positively affected by PL treatments (p≤ 0.05). We concluded that nebulization does not appear to meaningfully impact PL bacterial growth inhibition or protein composition. Further research is recommended to better characterize PL antimicrobial activity against clinically relevant pathogens and suitability for nebulization in horses.

ORGANISM(S): Equus Caballus

SUBMITTER: Kara Lascola

PROVIDER: PXD055023 | JPOST Repository | Thu Dec 26 00:00:00 GMT 2024

REPOSITORIES: jPOST

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			Action	DRS
	23-118-Lascola-Post-A1-120.raw	Raw
	23-118-Lascola-Post-B1-120.raw	Raw
	23-118-Lascola-Post-C1-120.raw	Raw
	23-118-Lascola-Pre-120_20231002154139.raw	Raw
	23-118-Lascola-platelet-lysate-Byonic-quan.mzML	Mzml

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Project description:<p>The study of antimicrobial resistance (AMR) in infectious diarrhea has generally been limited to cultivation, antimicrobial susceptibility testing and targeted PCR assays. When individual strains of significance are identified, whole genome shotgun (WGS) sequencing of important clones and clades is performed. Genes that encode resistance to antibiotics have been detected in environmental, insect, human and animal metagenomes and are known as "resistomes". While metagenomic datasets have been mined to characterize the healthy human gut resistome in the Human Microbiome Project and MetaHIT and in a Yanomani Amerindian cohort, directed metagenomic sequencing has not been used to examine the epidemiology of AMR. Especially in developing countries where sanitation is poor, diarrhea and enteric pathogens likely serve to disseminate antibiotic resistance elements of clinical significance. Unregulated use of antibiotics further exacerbates the problem by selection for acquisition of resistance. This is exemplified by recent reports of multiple antibiotic resistance in Shigella strains in India, in Escherichia coli in India and Pakistan, and in nontyphoidal Salmonella (NTS) in South-East Asia. We propose to use deep metagenomic sequencing and genome level assembly to study the epidemiology of AMR in stools of children suffering from diarrhea. Here the epidemiology component will be surveillance and analysis of the microbial composition (to the bacterial species/strain level where possible) and its constituent antimicrobial resistance genetic elements (such as plasmids, integrons, transposons and other mobile genetic elements, or MGEs) in samples from a cohort where diarrhea is prevalent and antibiotic exposure is endemic. The goal will be to assess whether consortia of specific mobile antimicrobial resistance elements associate with species/strains and whether their presence is enhanced or amplified in diarrheal microbiomes and in the presence of antibiotic exposure. This work could potentially identify clonal complexes of organisms and MGEs with enhanced resistance and the potential to transfer this resistance to other enteric pathogens.</p> <p>We have performed WGS, metagenomic assembly and gene/protein mapping to examine and characterize the types of AMR genes and transfer elements (transposons, integrons, bacteriophage, plasmids) and their distribution in bacterial species and strains assembled from DNA isolated from diarrheal and non-diarrheal stools. The samples were acquired from a cohort of pediatric patients and controls from Colombia, South America where antibiotic use is prevalent. As a control, the distribution and abundance of AMR genes can be compared to published studies where resistome gene lists from healthy cohort sequences were compiled. Our approach is more epidemiologic in nature, as we plan to identify and catalogue antimicrobial elements on MGEs capable of spread through a local population and further we will, where possible, link mobile antimicrobial resistance elements with specific strains within the population.</p>

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Characterization of Equine Platelet Lysate for Nebulization in the Horse

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