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. 2023 Nov 28;11(12):2877.
doi: 10.3390/microorganisms11122877.

In Vitro Efficacy of Isobutyl Cyanoacrylate Nanoparticles against Fish Bacterial Pathogens and Selection Preference by Rainbow Trout (Oncorhynchus mykiss)

Mawuko G Ahiable  1 , Kouki Matsunaga  1 , Mao Hokin  1 , Kazuhiro Iida  2 , Fumiaki Befu  2 , Syun-Ichirou Oshima  1
Affiliations

In Vitro Efficacy of Isobutyl Cyanoacrylate Nanoparticles against Fish Bacterial Pathogens and Selection Preference by Rainbow Trout (Oncorhynchus mykiss)

Mawuko G Ahiable et al. Microorganisms. .

Abstract

The upsurge in havoc being wreaked by antibiotic-resistant bacteria has led to an urgent need for efficacious alternatives to antibiotics. This study assessed the antibacterial efficacy of two isobutyl cyanoacrylate nanoparticles (iBCA-NPs), D6O and NP30, against major bacterial pathogens of fish. In vivo tests on rainbow trout were preceded by in vitro tests of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). NP30 exhibited higher efficacy than D60, but both iBCA-NPs demonstrated dose-dependent and species-specific in vitro antibacterial properties against the bacterial isolates. Generally, Gram-negative bacteria were more resistant to the iBCA-NPs. Streptococcus iniae, Tenacibaculum maritimum, and Photobacterium damselae were particularly sensitive to both iBCA-NPs. Administered to rainbow trout at 3571.4 mg (iBCA-NP)/kg feed, the iBCA-NPs produced a relative gain rate and survival rates comparable to the control (p > 0.05). The condition factor and the hepatosomatic and viscerosomatic indices of fish were indifferentiable (p > 0.05) between the iBCA-NP groups and the control. The iBCA-NPs caused no alteration in stress, oxidative stress (superoxide dismutase, SOD), plasma complement titer, or lysozyme activity. This study presents the first report of antibacterial activity of iBCA-NPs against Gram-negative bacteria. The results of this study suggest that D60 and NP30 may contribute to reducing the amounts of antibiotics and chemotherapeutic agents used in aquaculture.

Keywords: antibacterial efficacy; isobutyl cyanoacrylate nanoparticles; rainbow trout; safety.

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Conflict of interest statement

Befu Fumiaki and Kazuhiro Iida are employees of Chikami Miltec Inc. The remaining authors declare no conflict of interest.

Figures

Figure 1
Figure 1
(a) Preparation of the isobutyl cyanoacrylate nanoparticles (iBCA-NPs) used in this study. (b) Plate comparing cloudiness of the iBCA-NP solutions used in this study.
Figure 2
Figure 2
A plot to determine MIC, showing bacterial growth inhibition after 24 h in NP30 (a) vs. bacterial growth in dispersant (b). From the plot, the MIC of NP30 against E. tarda (typical) is 1.25 mg/mL.
Figure 3
Figure 3
Percent growth inhibition (GI %) of Vibrio spp. at all growth inhibiting concentrations: (a) GI % of Vibrio anguillarum for D60 (I) and NP30 (II); (b) GI % of Vibrio harveyi for D60 (I) and NP30 (II); (c) GI % of Vibrio parahaemolyticus for D60 (I) and NP30 (II); (d) GI % of Vibrio rotiferanus for D60 (I) and NP30 (II). All data are expressed as means ± SEM (n = 3).
Figure 3
Figure 3
Percent growth inhibition (GI %) of Vibrio spp. at all growth inhibiting concentrations: (a) GI % of Vibrio anguillarum for D60 (I) and NP30 (II); (b) GI % of Vibrio harveyi for D60 (I) and NP30 (II); (c) GI % of Vibrio parahaemolyticus for D60 (I) and NP30 (II); (d) GI % of Vibrio rotiferanus for D60 (I) and NP30 (II). All data are expressed as means ± SEM (n = 3).
Figure 4
Figure 4
Percent growth inhibition (GI %) of some Gram-positive bacteria strains at all growth-inhibiting concentrations observed: (a) GI % of Lactococcus garvieae; (b) GI % of Nocardia seriolae; (c) GI % of Streptococcus iniae for D60 (I) and NP30 (II). All data are expressed as means ± SEM (n = 3).
Figure 5
Figure 5
Percent growth inhibition (GI %) of some Gram-negative bacteria strains at all growth-inhibiting concentrations observed: (a) GI % of Photobacterium damselae subsp. piscicida; (b) GI % of Tericibaculum maritimum; (c) GI % of Aeromonas. salmonicida subsp. salmonicida; (d) GI % of Edwardsiella tarda (typical); (e) GI % of Edwardsiella tarda (atypical) for D60 (I) and NP30 (II). All data are expressed as means ± SEM (n = 3).
Figure 5
Figure 5
Percent growth inhibition (GI %) of some Gram-negative bacteria strains at all growth-inhibiting concentrations observed: (a) GI % of Photobacterium damselae subsp. piscicida; (b) GI % of Tericibaculum maritimum; (c) GI % of Aeromonas. salmonicida subsp. salmonicida; (d) GI % of Edwardsiella tarda (typical); (e) GI % of Edwardsiella tarda (atypical) for D60 (I) and NP30 (II). All data are expressed as means ± SEM (n = 3).
Figure 6
Figure 6
Overall bacterial sensitivity via percent growth inhibition by (A) D60 and (B) NP30 at 0.5 mg/mL in descending order.
Figure 6
Figure 6
Overall bacterial sensitivity via percent growth inhibition by (A) D60 and (B) NP30 at 0.5 mg/mL in descending order.
Figure 7
Figure 7
Fish growth and safety parameters. (a) Relative gain rate of rainbow trout for the study period. (b) Blood plasma lysozyme activity at OD 493 nm. (c) Plasma complement activity at OD 493 nm. (d) Plasma glucose concentrations. (e) Rate of plasma SOD inhibition (%) per treatment. All data are expressed as means ± SEM, where n = the number of replicates per treatment and N = the total number of fish for each test. Different letter superscripts, x, y, z, and asterisks (*) indicate significant differences between treatments and or sampling time (p < 0.05). CT = control group; M1, M2, and M3 = months 1, 2, and 3.
Figure 7
Figure 7
Fish growth and safety parameters. (a) Relative gain rate of rainbow trout for the study period. (b) Blood plasma lysozyme activity at OD 493 nm. (c) Plasma complement activity at OD 493 nm. (d) Plasma glucose concentrations. (e) Rate of plasma SOD inhibition (%) per treatment. All data are expressed as means ± SEM, where n = the number of replicates per treatment and N = the total number of fish for each test. Different letter superscripts, x, y, z, and asterisks (*) indicate significant differences between treatments and or sampling time (p < 0.05). CT = control group; M1, M2, and M3 = months 1, 2, and 3.
Figure 8
Figure 8
Plate comparing healthy rainbow trout (A,B) and one with a tumor (C,D) isolated from an NP30 (N = 1; where N = total number of fish).
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