Campylobacter Bacteriophage Cocktail Design Based on an Advanced Selection Scheme

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Abstract

Campylobacteriosis is a worldwide-occurring disease and has been the most commonly reported zoonotic gastrointestinal infection in the European Union in recent years. The development of successful phage-based intervention strategies will require a better understanding of phage–bacteria interactions to facilitate advances in phage cocktail design. Therefore, this study aimed to investigate the effects of newly isolated group II and group III phages and their combinations on current Campylobacter field strains. A continuous workflow for host range and efficiency of plating (EOP) value determination was combined with a qPCR-based phage group identification and a liquid-based planktonic killing assay (PKA). An advanced analysis scheme allowed us to evaluate phage cocktails by their efficacy in inhibiting bacterial population growth and the resulting phage concentrations. The results of this study indicate that data obtained from PKAs are more accurate than host range data based on plaque formation (EOP). Planktonic killing assays with Campylobacter appear to be a useful tool for a straightforward cocktail design. Results show that a group II phage vB_CcM-LmqsCP218-2c2 and group III phage vB_CjM-LmqsCP1-1 mixture would be most promising for practical applications against Campylobacter coli and Campylobacter jejuni.

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The European Union One Health 2019 Zoonoses Report

(2021)

Abstract This report of the EFSA and the European Centre for Disease Prevention and Control presents the results of zoonoses monitoring activities carried out in 2019 in 36 European countries (28 Member States (MS) and eight non‐MS). The first and second most reported zoonoses in humans were campylobacteriosis and salmonellosis, respectively. The EU trend for confirmed human cases of these two diseases was stable (flat) during 2015–2019. The proportion of human salmonellosis cases due to Salmonella Enteritidis acquired in the EU was similar to that in 2017–2018. Of the 26 MS reporting on Salmonella control programmes in poultry, 18 met the reduction targets, whereas eight failed to meet at least one. The EU prevalence of Salmonella target serovar‐positive flocks has been stable since 2015 for breeding hens, laying hens, broilers and fattening turkeys, with fluctuations for breeding turkey flocks. Salmonella results from competent authorities for pig carcases and for poultry tested through national control programmes were more frequently positive than those from food business operators. Shiga toxin‐producing Escherichia coli (STEC) infection was the third most reported zoonosis in humans and increased from 2015 to 2019. Yersiniosis was the fourth most reported zoonosis in humans in 2019 with a stable trend in 2015–2019. The EU trend of confirmed listeriosis cases remained stable in 2015–2019 after a long period of increase. Listeria rarely exceeded the EU food safety limit tested in ready‐to‐eat food. In total, 5,175 food‐borne outbreaks were reported. Salmonella remained the most detected agent but the number of outbreaks due to S. Enteritidis decreased. Norovirus in fish and fishery products was the agent/food pair causing the highest number of strong‐evidence outbreaks. The report provides further updates on bovine tuberculosis, Brucella, Trichinella, Echinococcus, Toxoplasma, rabies, West Nile virus, Coxiella burnetii (Q fever) and tularaemia.

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Campylobacter.

John E Moore, Deborah Corcoran, James Dooley … (2015)

Species within the genus, Campylobacter, have emerged over the last three decades as significant clinical pathogens, particularly of human public health concern, where the majority of acute bacterial enteritis in the Western world is due to these organisms. Of particular concern are the species, C. jejuni and C. coli, which are responsible for most of these gastrointestinal-related infections. Although these organisms have already emerged as causative agents of zoonoses, several aspects of their epidemiology and pathophysiology are only beginning to emerge. Trends in increasing antibiotic resistance are beginning to emerge with oral antibiotics, which may be the drug of choice for when it is necessary to intervene chemotherapeutically. This review wishes to examine (i) emerging clinical aspects of the disease, such as Guillain Barre syndrome (GBS), (ii) the association between these organisms and poultry as a natural host, (iii) environmental aspects of Campylobacter epidemiology, (iv) the emergence of atypical campylobacters (v) emerging trends in antibiotic resistance, (vi) adoption of modern methods for the detection of campylobacters.

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A Method for Generation Phage Cocktail with Great Therapeutic Potential

Jingmin Gu, Xiaohe Liu, Yue Li … (2012)

Background Bacteriophage could be an alternative to conventional antibiotic therapy against multidrug-resistant bacteria. However, the emergence of resistant variants after phage treatment limited its therapeutic application. Methodology/Principal Findings In this study, an approach, named “Step-by-Step” (SBS), has been established. This method takes advantage of the occurrence of phage-resistant bacteria variants and ensures that phages lytic for wild-type strain and its phage-resistant variants are selected. A phage cocktail lytic for Klebsiella pneumoniae was established by the SBS method. This phage cocktail consisted of three phages (GH-K1, GH-K2 and GH-K3) which have different but overlapping host strains. Several phage-resistant variants of Klebsiella pneumoniae were isolated after different phages treatments. The virulence of these variants was much weaker [minimal lethal doses (MLD)>1.3×109 cfu/mouse] than that of wild-type K7 countpart (MLD = 2.5×103 cfu/mouse). Compared with any single phage, the phage cocktail significantly reduced the mutation frequency of Klebsiella pneumoniae and effectively rescued Klebsiella pneumoniae bacteremia in a murine K7 strain challenge model. The minimal protective dose (MPD) of the phage cocktail which was sufficient to protect bacteremic mice from lethal K7 infection was only 3.0×104 pfu, significantly smaller (p<0.01) than that of single monophage. Moreover, a delayed administration of this phage cocktail was still effective in protection against K7 challenge. Conclusions/Significance Our data showed that the phage cocktail was more effective in reducing bacterial mutation frequency and in the rescue of murine bacteremia than monophage suggesting that phage cocktail established by SBS method has great therapeutic potential for multidrug-resistant bacteria infection.