Fig 1.
Viable ɸV10 recovered from printing onto dissolvable paper.
Five stock concentrations of ɸV10 were printed onto 1cm2 pieces of dissolvable paper. Upon dissolution and dilution, the illustrated number of viable ɸV10 were recovered. The left column for each treatment group represents a theoretical 100% recovery, whereas the right column describes the number of viable phage experimentally recovered.
Fig 2.
Recovery of ɸV10 applied to dissolvable paper in different deposition patterns.
(A) Deposition patterns of ɸV10 onto 1cm2 dissolvable paper. A consistent volume (1.75 μL) of liquid phage stock applied to papers in three patterns. i) ɸV10 printed onto dissolvable paper across the entire surface of the paper using 10nL droplets in a grid-like pattern of deposition (surface area of 100 mm2). ii) Five droplets containing 350 nL were pipetted onto the paper in approximately equal spacing (surface area average of 21.24 ± 2.94 mm2, n = 15). iii) One droplet containing 1.75 μL was pipetted in the center of the dissolvable paper (surface area average of 17.78 ± 2.23 mm2, n = 15). (B) Recovery of ɸV10 applied to dissolvable paper in different deposition patterns. Deposition patterns correspond to the 1.75 μL patterns listed in Fig 2A. Significant differences between recovered PhiV10 from various deposition patterns onto dissolvable papers are a result of a one-way ANOVA analysis with post-hoc Duncan test. Capital letters denote significantly distinct (p < 0.05) titer values as a result. Distinct differences in recovered viable phage counts are seen and are inversely proportional to the surface area of phage applied to papers. The control group were 1.75 μL of the same phage stock into 1mL sterile water. Results shown are from three replicates of each treatment.
Table 1.
Viable ɸV10 phage recovered from dissolvable papers after varying time of drying.
Fig 3.
Longevity of ɸV10 on dissolvable papers stored at 4°C.
Data shown include one-way ANOVA analysis to show significant (p < 0.05) differences between treatment groups at each timepoint and within each treatment group over time. Lowercase letters denote significant differences between treatments within a single timepoint. For each timepoint, the lowercase letter “a” corresponds to the highest, statistically distinct value for recovered phage titer, with “b” representing the second highest, statistically distinct value, etc. Capital letters denote significant differences within a single treatment group over time. Capital letters are only significant within a single treatment group over time and are assigned in order of descending titers (e.g. “A” is always the largest, significantly different titer for a single treatment group).
Fig 4.
Longevity of lyophilized ɸV10.
Titers from treatment groups of ɸV10 stored in BPB with varying concentrations of sucrose with or without lyophilization. One-way ANOVA analysis has been applied to illustrate significant differences between viable phage titers between treatment groups over time. Each lowercase data label corresponds to a significantly different (p < 0.05) titer value within each timepoint. These lowercase labels signify statistically distinct titer values between the different treatments only within a single timepoint. Capital letters denote significantly distinct (p < 0.05) titer values for a single treatment and time point compared to each other time point within the same treatment conditions. These capital data labels correspond to how recovery of phage from a single treatment changed over time; each capital letter denotes a statistically distinct titer value for a single treatment over several timepoints.
Fig 5.
ɸV10nLuc titers after lyophilization and storage at 23°C.
(A) Initial ɸV10nLuc titers after lyophilization. For each lyophilized treatment, titers were compared to their respective liquid buffer controls. Asterisks denote statistical significance between groups (p < 0.05). (B) ɸV10nLuc titers after lyophilization and 18 days of storage at 23°C. For each lyophilized treatment, titers were compared to their respective liquid buffer controls. Asterisks denote statistical significance between groups (p < 0.05).
Fig 6.
Rates of lysogeny for ɸV10nLuc in liquid and lyophilized buffers after 18 days.
The rates of lysogeny are determined by averaging triplicate experiments where the number of kanamycin resistant lysogens that are formed for a given phage sample are divided by the total number of phage in the sample determined by plaque assays. Capital letters denote a sample’s statistical grouping compared to the phage buffer control group. Lowercase letters denote significant differences between all treatment groups (e.g. 0.1M sucrose buffer and lyophilized 0.1M sucrose buffer samples).