Potential of lactoferrin to prevent antibiotic-induced Clostridium difficile infection.

C H Chilton,M Brindell,M H Wilcox,K Śpiewak,G S Crowther,G Singh,T M Monaghan

doi:10.1093/jac/dkv452

Abstract

ObjectivesClostridium difficile infection (CDI) is a global healthcare problem. Recent evidence suggests that the availability of iron may be important for C. difficile growth. This study evaluated the comparative effects of iron-depleted (1% Fe3+ saturated) bovine apo-lactoferrin (apo-bLf) and iron-saturated (85% Fe3+ saturated) bovine holo-lactoferrin (holo-bLf) in a human in vitro gut model that simulates CDI.MethodsTwo parallel triple-stage chemostat gut models were inoculated with pooled human faeces and spiked with C. difficile spores (strain 027 210, PCR ribotype 027). Holo- or apo-bLf was instilled (5 mg/mL, once daily) for 35 days. After 7 days, clindamycin was instilled (33.9 mg/L, four times daily) to induce simulated CDI. Indigenous microflora populations, C. difficile total counts and spores, cytotoxin titres, short chain fatty acid concentrations, biometal concentrations, lactoferrin concentration and iron content of lactoferrin were monitored daily.ResultsIn the apo-bLf model, germination of C. difficile spores occurred 6 days post instillation of clindamycin, followed by rapid vegetative cell proliferation and detectable toxin production. By contrast, in the holo-bLf model, only a modest vegetative cell population was observed until 16 days post antibiotic administration. Notably, no toxin was detected in this model. In separate batch culture experiments, holo-bLf prevented C. difficile vegetative cell growth and toxin production, whereas apo-bLf and iron alone did not.ConclusionsHolo-bLf, but not apo-bLf, delayed C. difficile growth and prevented toxin production in a human gut model of CDI. This inhibitory effect may be iron independent. These observations suggest that bLf in its iron-saturated state could be used as a novel preventative or treatment strategy for CDI.

Highlights

Clostridium difficile infection (CDI) is a global healthcare problem
As seen in previous gut-model experiments, C. difficile remained as spores before and during clindamycin instillation in both models (Figure 2)
In the apo-Bovine lactoferrin (bLf) model, this germination was followed by rapid vegetative cell proliferation and toxin production (Figure 2a)

Summary

Introduction

Clostridium difficile infection (CDI) is a global healthcare problem. It is the leading cause of hospital-acquired infectious diarrhoea, causing significant morbidity and societal financial burden.[1,2] Recurrent infections and increasing antibiotic resistance have complicated the treatment of CDI.[3,4,5] There is an urgent need for the development of novel, non-antibioticbased therapeutic and preventative strategies. To successfully sustain an infection, most bacteria, fungi and protozoa require a continuous supply of host iron, which participates in many essential metabolic processes.[6] Recent in vitro evidence suggests that the availability and source of iron may be important for C. difficile growth.[7] The development of pharmaceutical agents that can manipulate microbial access to iron may help prevent and possibly treat CDI

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