Determination of anti-phage antibodies in calf sera following application of Escherichia coli and Mannheimia haemolytica-specific bacteriophages

Urban-Chmiel, Renata; Nowaczek, Anna; Chmiel, Paulina; Wawrzykowski, Jacek; Pyzik, Ewelina; Marek, Agnieszka; Stępien-Pyśniak, Dagmara; Alomari, Mohammed M.M.; Dec, Marta

doi:10.2478/jvetres-2022-0041

Figures & Tables

Sodium dodecyl sulphate–polyacrylamide gel electrophoresis electropherograms with phage proteinsA: Lines 1, 2 and 3 – 25 A2 and A5 phages specific for M. haemolytica strains; B: Lines 1, 2 and 3 –26, 27 and 29 phages specific for E. coli
M –10–240 kDa molecular weight protein ladder — Sodium dodecyl sulphate–polyacrylamide gel electrophoresis electropherograms with phage proteinsA: Lines 1, 2 and 3 – 25 A2 and A5 phages specific for M. haemolytica strains; B: Lines 1, 2 and 3 –26, 27 and 29 phages specific for E. coli M –10–240 kDa molecular weight protein ladder

Two-dimensional electrophoresis of phage proteinsA – phages specific for M. haemolytica; B – phages specific for E. coli
M – 7–240 kDa molecular weight standard — Two-dimensional electrophoresis of phage proteinsA – phages specific for M. haemolytica; B – phages specific for E. coli M – 7–240 kDa molecular weight standard

Average absorbance values (±standard deviation) measured in serum samples from calves by ELISA in response to application of bacteriophages specific for E_ coli and M_ haemolytica

Parameter	FBS			Calves with clinical signs of disease after phage application			Healthy calves after phage application			Healthy calves not treated with phages
Parameter
	IgG	IgM	IgA	IgG	IgM	IgA	IgG	IgM	IgA	IgG	IgM	IgA
Mean level of antibodies against whole M. haemolytica and E. coli bacteriophages	0.007 ± 0.004	0.008 ± 0.006	0.001 ± 0.0009	0.64 ± 0.09^ab	0.192 ± 0.068^ab	0.114 ± 0.048^ab	0.845 ± 0.111^ab	0.239 ± 0.082^ab	0.021 ± 0.007^a	0.056 ± 0.024^a	0.019 ± 0.00^a	0.017 ± 0.01^a
Mean level of antibodies against whole E. coli phages	0.0032 ± 0.001	0.014 ± 0.008	0.001 ± 0.0008	0.74 ± 0.16^ab	0.211 ± 0.052^ab	0.19 ± 0.09^ab	0.71 ± 0.13^ab	0.203 ± 0.2^ab	0.029 ± 0.003^a	0.06 ± 0.009^a	0.096 ± 0.038^a	0.02 ± 0.007^a
Mean level of antibodies against whole M. haemolytica phages	0.005 ± 0.004	0.004 ± 0.0001	0.002 ± 0.0005	0.95 ± 0.08^ab	0.99 ± 0.111^ab	0.23 ± 0.005^ab	0.6 ± 0.1^a	0.33 ± 0.02^ab	0.02 ± 0.002^a	0.72 ± 0.017^a	0.08 ± 0.03^a	0.03 ± 0.001^a
Mean level of antibodies against M. haemolytica and E. coli extracted proteins	0.005 ± 0.003	0.009 ± 0.007	0.001 ± 0.0005	0.71 ± 0.134^ab	0.18 ± 0.34^ab	0.15 ± 0.05^ab	0.53 ± 0.13^ab	0.226 ± 0.13^ab	0.03 ± 0.016	0.06 ± 0.01^a	0.07 ± 0.04^a	0.02 ± 0.0009^a
Mean level of antibodies against E. coli extracted proteins	0.004 ± 0.003	0.002 ± 0.0009	0.001 ± 0.0001	0.83 ± 0.032^ab	0.21 ± 0.004^ab	0.201 ± 0.002^ab	0.68 ± 0.15^ab	0.21 ± 0.004^ab	0.099 ± 0.007^a	0.062 ± 0.001^a	0.042 ± 0.002^a	0.02 ± 0.008^a
Mean level of antibodies against M. haemolytica extracted proteins	0.007 ± 0.004	0.004 ± 0.002	0.001 ± 0.000	0.77 ± 0.13^ab	0.3 ± 0.04^ab	0.15 ± 0.03^ab	0.65 ± 0.21^ab	0.26 ± 0.18 ^ab	0.05 ± 0.02^a	0.071 ± 0.04^a	0.05 ± 0.02^a	0.03 ± 0.01^a

Average values obtained in the phage neutralisation assay

Type of phage solution	Mean K ± SD in untreated calves	Mean K ± SD in treated calves
E. coli phages	0.0019 ± 0.001	7.28 ± 0.54*
M. haemolytica phages	0.0046 ± 0.003	5.57 ± 1.81*
E. coli extracted proteins	0.00156 ± 0.0004	6.74 ± 0.5*
M. haemolytica extracted proteins	0.0013 ± 0.0005	5.053 ± 1.53*

Determination of anti-phage antibodies in calf sera following application of Escherichia coli and Mannheimia haemolytica-specific bacteriophages

Figures & Tables

Fig. 1

Fig. 2

Average absorbance values (±standard deviation) measured in serum samples from calves by ELISA in response to application of bacteriophages specific for E_ coli and M_ haemolytica

Average values obtained in the phage neutralisation assay

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