Salmonella stays one of the crucial vital foodborne bacterial pathogens worldwide, and is steadily linked with the consumption of contaminated poultry meat and eggs. Regardless of some famous successes in lowering specific serotypes and strains, different serotypes have turn into extra problematic. Progress has additionally been annoyed by rising resistance to antimicrobials and biocides.

R.J. Atterbury – Faculty of Veterinary Medication and Science, College of Nottingham, Sutton Bonington, UK

Organic management utilizing bacteriophages or predatory micro organism is another strategy to Salmonella management in poultry which has been efficient in laboratory, experimental and/or discipline trial settings. The European Union is at present contemplating authorizing the primary phage-based industrial product for the remedy of Salmonella in poultry. Questions stay concerning the regulation and industrial standing of such merchandise, in addition to their long-term efficacy; however their means to particularly goal multidrug resistant micro organism might present a brand new approach of lowering our reliance on antimicrobials and reduce the burden of illness in each animals and people sooner or later.

Introduction
Salmonellosis stays one of the crucial frequent food-borne zoonoses, constituting a worldwide main public well being concern. Though Salmonella might be acquired from a spread of meals, poultry meat and eggs stay outstanding (Antunes et al., 2016). Human salmonellosis circumstances within the US are estimated at 1.35 million per 12 months (Facilities for Illness Management, 2019). Likewise, circumstances within the UK and EU haven’t modified considerably over the previous decade.
Success in controlling some Salmonella serotypes (e.g. Enteritidis) in poultry by way of vaccination and biocontrol have been punctuated by failure to manage different serotypes. A current instance of this was an outbreak of Salmonella Infantis in the US in 2018, affecting 32 states (Facilities for Illness Management, 2019). Within the EU, Salmonella Infantis is now the 4^th most typical serotype from human infections, and essentially the most frequent serotype in broilers flocks (45.6 %) and broiler meat (50.6 %) in Europe (Alba et al., 2020). Extra worryingly, multidrug-resistant isolates have gotten more and more widespread in Europe, notably these which have acquired a pESI megaplasmid encoding resistance to a number of courses of antibiotics in addition to virulence genes and enhanced tolerance of heavy metals and biocides (Tyson et al., 2021).
One different strategy to Salmonella management is using organic management brokers; both viruses which particularly infect Salmonella – bacteriophages – or use of predatory micro organism reminiscent of Bdellovibrio bacteriovorus. In a current evaluate of alternate options to antibiotics, funded by the Wellcome Belief and UK Division of Well being, bacteriophage-based remedies comprised three of the ten Tier 1 (most promising) new applied sciences (Czaplewski et al., 2016). Though much less developed, Bdellovibrio has been used efficiently to manage an infection in a number of animal fashions of illness, together with the remedy of Salmonella in poultry (Atterbury et al., 2011).
Organic management has distinctive benefits over antimicrobials. These brokers are each self-replicating and self-limiting – reproducing solely when prone micro organism are current. In contrast to broad-spectrum antibiotics, they aim a particular genus, species or pressure of bacterium, avoiding potential dangerous dysbiosis within the affected person. Likewise, a number of research have discovered that Bdellovibrio can prey upon multidrug resistant pathogens in biofilms (Atterbury and Tyson, 2021; Solar et al., 2017). As well as, Bdellovibrio can survive for as much as 24 h inside eukaryotic cells with out adversarial results, suggesting that it may very well be utilized to manage pathogens with an intracellular element to their lifecycle, together with Salmonella (Atterbury and Tyson, 2021).

Technique
Candidate therapeutic bacteriophage are normally remoted from an setting the place their bacterial hosts are ample. Within the case of Salmonella, that is typically poultry feces, wastewater or human sewage. As soon as purified, these phages are screened towards a big panel of goal serotypes and strains which must be chosen to replicate the variety of pathogens affecting the inhabitants. Phages that infect a broad vary of micro organism, both individually or together (cocktail) are normally chosen for genome sequencing and evaluation. This step permits the identification of undesirable phage e.g. temperate phage which don’t underdo a direct and predictable lytic an infection cycle. It additionally identifies phage carrying undesirable genes reminiscent of these related to virulence. Extra detailed in vitro characterization of the phage could also be carried out in parallel, reminiscent of figuring out the burst dimension and replication kinetics of an infection. For promising candidates, the precise phage receptor(s) on the host bacterium could also be recognized, together with an evaluation of how steadily resistance to phage happens.
More and more, bioinformatics is getting used to facilitate the characterization and collection of phage. There are instruments accessible to foretell the taxonomy, DNA packaging mechanisms (Garneau et al., 2017), lifecycle (McNair et al., 2012), receptor specificity and host vary (Zhang et al., 2017) of candidate phage. As soon as a phage is characterised, on-line instruments can be utilized to match it with others uploaded to the database which can be helpful for cross-validation (Rangel-Pineros et al., 2021). Additional confirmatory exams can then be carried out within the laboratory, for instance to determine the phage receptors predicted in silico. Thereafter, the phage candidates might be examined for efficacy in easy animal fashions reminiscent of Galleria mellonella and their stability in industrial preparations earlier than taking ahead to experimental and discipline trials in poultry.
Bdellovibrio, like phage, is most frequently remoted from environments the place their prey are ample. That is typically in aquatic environments or sewage. In contrast to most bacteriophage, Bdellovibrio targets a number of genera of Gram-negative pathogens, and resistance to predation is uncommon and transient (Marine et al., 2020). There’s proof that Bdellovibrio or like organisms are current within the intestinal tracts of a spread of animal species, together with poultry (Schwudke et al., 2001). Nevertheless, these micro organism have been tough to isolate and exploit instantly. Bdellovibrio are metabolically energetic and chemotactic in direction of their prey. As such, in contrast to phage they don’t seem to be reliant on passive diffusion to unfold by way of a bacterial inhabitants. Nevertheless, it additionally means the choices accessible for therapeutic supply are extra restricted than with phage and industrial preparations could also be tougher.

Outcomes
Salmonella is steadily a goal of organic management trials owing each to its significance as a human pathogen and as a trigger of serious illness and manufacturing losses in livestock. Most effort has been directed in direction of controlling vital non-host-restricted serotypes of Salmonella, reminiscent of Enteritidis and Typhimurium that are extra important from a regulatory perspective within the EU and elsewhere.
Cocktails of lytic phage have been used to scale back intestinal carriage of Salmonella by between 1 and three.5 log₁₀ CFU/g (Atterbury et al., 2007; Sklar and Joerger, 2007). These reductions have been typically accompanied by marked enchancment or elimination of medical indicators of illness (Sklar and Joerger, 2007). Phage remedy has been mixed with seeder fashions of an infection to show how efficient phage might be in stopping horizontal an infection of chicks by Salmonella. Lim et al. (2012) confirmed that remedy with as much as 109 PFU/g of phage in feed may cut back intestinal colonization by as much as 1 log₁₀ CFU/g, whereas 70% of the contact chickens had no detectable Salmonella Enteritidis colonization. Henriques et al. (2013) considerably diminished the unfold of Salmonella Enteritidis in chicks by treating fertilized eggs with a cocktail of two phages. Medical indicators of illness on the finish of the eight-day research weren’t considerably completely different from the management group.
Extra just lately, two research have examined the impression of making use of therapeutic phage on the hen microbiome. Kosznik-Kwaśnicka et al. (2022) discovered that each phage and antibiotics (enrofloxacin and colostin) have been all in a position to cut back Salmonella to under detectable ranges in contaminated chickens. Nevertheless, antibiotic use was related to important and extended modifications within the microbiome whereas the results of phage have been transient and normalized a number of weeks after remedy. Equally, Clavijo et al. (2022) used a proprietary Salmonella phage product in a industrial broiler farm to evaluate the impact on the intestine microbiome. No important results of phage on the maturation of the microbiome have been detected.
Bdellovibrio has been remoted beforehand from the intestinal tracts of chickens in addition to people and different vertebrates (Schwudke et al., 2001). Atterbury et al. (2011) used Bdellovibrio to deal with chickens experimentally contaminated with Salmonella Enteritidis. Important reductions of over 1 log₁₀ CFU/g have been recorded within the ceca of handled birds in contrast with buffer-treated controls. Bdellovibrio-treated birds additionally had considerably fewer cecal abnormalities in contrast with each untreated controls and birds handled with a non-predatory Bdellovibrio mutant. Apparently, in vitro experiments utilizing each phage and Bdellovibrio confirmed that the mixture of each virus and predator was in a position to eradicate pathogen populations rather more successfully than both used independently (Hobley et al., 2020)

Dialogue
Bacteriophages have been proven to considerably cut back the intestinal carriage of a number of serotypes of Salmonella in broiler and layer chickens. These reductions have been steadily accompanied by enhancements in medical manifestations of illness (diminished perihepatitis, pericarditis, typhlitis) in addition to feed conversion ratios. Though phage-resistant variants of Salmonella have been recorded, these phenotypes are sometimes transient, and related to diminished health in animal fashions of an infection. Resistance to Bdellovibrio then again is extraordinarily uncommon, and solely conferred by the presence of an intact S-layer which is metabolically costly to keep up. Apparently, when Bdellovibrio and bacteriophage are utilized in mixture, they will act synergistically for a larger discount within the goal pathogen with out the event of resistance.
Regardless of their promise, there stay some technical, industrial and regulatory hurdles which should be overcome earlier than bacteriophages and Bdellovibrio may very well be used extra extensively within the UK/EU. These brokers don’t match simply inside present regulatory frameworks which have been designed for outlined chemotherapeutics. Likewise, the flexibility to guard mental property referring to naturally-occurring organic entities is an open query. Regardless of these obstacles, industrial phage merchandise focusing on food-borne pathogens can be found in the US and the EU is in superior discussions to authorize the primary bacteriophage remedy for livestock within the bloc. Given the urgent want to manage multidrug-resistant infections, and the lowering efficacy and availability of antibiotics to deal with them, organic management brokers reminiscent of Bdellovibrio and bacteriophage supply a possible resolution.

References can be found on request
From the Proceedings of the Australian Poultry Science Symposium 2023