In this research paper the in vitro antimicrobial and antibiofilm activity of phage cocktail against the coagulase negative Mammaliicoccus sciuri was investigated. Three M. sciuri isolates obtained from clinical bovine mastitis samples were characterized and identified by 16S rRNA gene sequencing. Bacteriophages with lytic activity against M. sciuri isolates were isolated from dairy farm effluents. Two typical phages were isolated using standard enrichment and plaque assay techniques, purified by polyethylene glycol precipitation, and morphologically characterized based on shape and size using transmission electron microscopy. This was followed by determination of host range using spot tests and stability to varying temperature, pH and UV treatment. The phage cocktail suppressed bacterial activity within 30 min of exposure. Crystal violet assay showed that the tested phages and their cocktail significantly reduced the biofilm biomass of all three M. sciuri strains compared to the untreated control in vitro within 24 h with a single dosing. Transmission electron micrography of the purified phage particle revealed an icosahedral head and a rigid contractile tail, characteristic of the class Caudoviricetes. The findings open new avenues in phage-based antimicrobial approaches for controlling contagious and teat skin opportunistic bacteria causing bovine mastitis.

The performance of the starter culture is a critical factor that decides the quality of fermented milk. Dahi is a fermented milk product popular in India made using a mixed starter culture of lactic acid bacteria comprising acid and flavour producers. The prevalence of bacteriophages in the dairy environment can critically affect the activity of these starter cultures resulting in starter failure. As there is little information available on the occurrence of bacteriophages in the dairy environment of Kerala, this research communication examines the presence of lytic bacteriophages acting against three potential flavour-producing strains of Lacticaseibacillus paracasei (Lc. paracasei). Dairy effluent samples were screened for the presence of phages against the strains of Lc. paracasei by the multiple host enrichment method. Plates showing clearance zone in spot assay were confirmed for the presence of phages by double-layer agar assay. The plaques obtained in the double-layer agar assay were purified for further identification by next-generation sequencing. A bacteriophage infecting one of the three strains of Lc. paracasei was detected by the plaque assay and the blast annotation of the bacteriophage sequence found 86.05% similarity of the phage to Siphoviridae family. The study endorses the need for monitoring phages in the dairy environment to control phage-related starter failure in the state of Kerala.

Two experiments were conducted to investigate the effects of dietary supplementation of bacteriophage cocktail, probiotics and a combination of these two supplements on performance and gut health of weanling pigs. In Experiment 1, 150 weaned piglets were randomly allotted to three treatments on the basis of BW. The dietary treatments included a basal diet supplemented with 0 (control), 1.0 and 1.5 g/kg bacteriophage cocktail. Pigs fed 1.0 and 1.5 g/kg bacteriophage product had greater (P<0.05) average daily gain (ADG), apparent total tract digestibility of dry matter from day 22 to 35, ileal Lactobacillus spp., villus height (duodenum and jejunum), and fewer coliforms (ileum) and Clostridium spp. (ileum). In Experiment 2, 200 weaned piglets were randomly allotted to four treatments. Dietary treatments included basal diet, basal diet supplemented with 3.0 g/kg fermented probiotic product (P), 1.0 g/kg bacteriophage cocktail (B) and combination of 1.0 g/kg bacteriophage cocktail and 3.0 g/kg fermented probiotic product. Pigs fed bacteriophage cocktail diets had greater (P<0.05) overall ADG, gain to feed ratio (G : F), fecal score from day 8 to day 21, and pigs fed bacteriophage cocktail diets had fewer coliforms (ileum) Clostridium spp. (ileum and cecum). Probiotics significantly increased G : F, colonization of Lactobacillus spp. in ileum. At day 35, bacteriophage treatment group showed greater (P<0.05) villus height of the duodenum, but a deeper crypt in duodenum. The present results indicate that the bacteriophage cocktail had a potential to enhance the performance and gut health of weanling pigs, however their combination with probiotics did not show an interaction.

Antibiotics are one of the most important medical discoveries of the 20th century and will remain an essential tool for treating animal and human diseases in the 21st century. However, antibiotic resistance among bacterial pathogens and concerns over their extensive use in food animals has garnered global interest in limiting antibiotic use in animal agriculture. Yet, limiting the availability of medical interventions to prevent and control animal diseases on the farm will directly impact global food security and safety as well as animal and human health. Insufficient attention has been given to the scientific breakthroughs and novel technologies that provide alternatives to antibiotics. The objectives of the symposium ‘Alternatives to Antibiotics’ were to highlight promising research results and novel technologies that could potentially lead to alternatives to conventional antibiotics, and assess challenges associated with their commercialization, and provide actionable strategies to support development of alternative antimicrobials. The symposium focused on the latest scientific breakthroughs and technologies that could provide new options and alternative strategies for preventing and treating diseases of animals. Some of these new technologies have direct applications as medical interventions for human health, but the focus of the symposium was animal production, animal health and food safety during food-animal production. Five subject areas were explored in detail through scientific presentations and expert panel discussions, including: (1) alternatives to antibiotics, lessons from nature; (2) immune modulation approaches to enhance disease resistance and to treat animal diseases; (3) gut microbiome and immune development, health and diseases; (4) alternatives to antibiotics for animal production; and (5) regulatory pathways to enable the licensure of alternatives to antibiotics.

This paper addresses food safety in beef cattle production, with particular emphasis on factors that affect the prevalence of Escherichia coli O157:H7 in beef cattle and on control methods that have been investigated. Product recalls and foodborne diseases due to this organism continue to occur even though control measures have been under investigation for over 20 years. Most meatborne outbreaks are due to improper food handling practices and consumption of undercooked meat. However, the majority of pathogenic bacteria that can spread at slaughter by cross-contamination can be traced back to the farm rather than originating from the slaughter plant. This would ideally require the adoption of rigorous on-farm intervention strategies to mitigate risks at the farm level. On-farm strategies to control and reduce E. coli O157:H7 at the farm level will reduce the risk of carcass contamination at slaughter and processing facilities although they will not eliminate E. coli O157:H7. The most successful strategy for reducing the risk of contamination of beef and beef products will involve the implementation of both pre- and post-harvest measures.

We construct a stochastic model ofbacteriophage parasitism of a host bacteria that accounts fordemographic stochasticity of host and parasite and allows formultiple bacteriophage adsorption to host. We analyze the associateddeterministic model, identifying the basic reproductive number forphage proliferation, showing that host and phage persist when itexceeds unity, and establishing that the distribution of adsorbedphage on a host is binomial with slowly evolving mean. Notsurprisingly, extinction of the parasite or both host and parasitecan occur for the stochastic model.

The successful use of virulent (lytic) bacteriophages (phages) in preventing and treating neonatal enterotoxigenic Escherichia coli infections in calves, lambs and pigs has prompted investigation of other applications of phage therapy in food animals. While results have been very variable, some indicate that phage therapy is potentially useful in virulent Salmonella and E. coli infections in chickens, calves and pigs, and in control of the food-borne pathogens Salmonella and Campylobacter jejuni in chickens and E. coli O157:H7 in cattle. However, more rigorous and comprehensive research is required to determine the true potential of phage therapy. Particular challenges include the selection and characterization of phages, practical modes of administration, and development of formulations that maintain the viability of phages for administration. Also, meaningful evaluation of phage therapy will require animal studies that closely represent the intended use, and will include thorough investigation of the emergence and characteristics of phage resistant bacteria. As well, effective use will require understanding the ecology and dynamics of the endemic and therapeutic phages and their interactions with target bacteria in the farm environment. In the event that the potential of phage therapy is realized, adoption will depend on its efficacy and complementarity relative to other interventions. Another potential challenge will be regulatory approval.