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. 2022 Mar 2;9(3):109.
doi: 10.3390/vetsci9030109.

Genetic Relatedness, Antibiotic Resistance, and Effect of Silver Nanoparticle on Biofilm Formation by Clostridium perfringens Isolated from Chickens, Pigeons, Camels, and Human Consumers

Heba A Ahmed  1 , Rasha M El Bayomi  2 , Rehab I Hamed  3 , Rasha A Mohsen  4 , Fatma A El-Gohary  5 , Ahmed A Hefny  6 , Eman Elkhawaga  7 , Hala M N Tolba  8
Affiliations

Genetic Relatedness, Antibiotic Resistance, and Effect of Silver Nanoparticle on Biofilm Formation by Clostridium perfringens Isolated from Chickens, Pigeons, Camels, and Human Consumers

Heba A Ahmed et al. Vet Sci. .

Abstract

In this study, we determined the prevalence and toxin types of antibiotic-resistant Clostridium perfringens in chicken, pigeons, camels, and humans. We investigated the inhibitory effects of AgNPs on biofilm formation ability of the isolates and the genetic relatedness of the isolates from various sources determined using RAPD-PCR. Fifty isolates were identified using PCR, and all the isolates were of type A. The cpe and cpb2 genes were detected in 12% and 56% of the isolates, respectively. The effect of AgNPs on biofilm production of six representative isolates indicated that at the highest concentration of AgNPs (100 µg/mL), the inhibition percentages were 80.8-82.8%. The RAPD-PCR patterns of the 50 C. perfringens isolates from various sources revealed 33 profiles and four clusters, and the discriminatory power of RAPD-PCR was high. Multidrug-resistant C. perfringens isolates are predominant in the study area. The inhibition of biofilm formation by C. perfringens isolates was dose-dependent, and RAPD-PCR is a promising method for studying the genetic relatedness between the isolates from various sources. This is the first report of AgNPs' anti-biofilm activity against C. perfringens from chickens, pigeons, camels, and humans, to the best of our knowledge.

Keywords: Clostridium perfringens; RAPD-PCR genotyping; biofilm inhibition; silver nanoparticles; toxinotyping.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Multifocal pale foci of mucosal necrosis and the lumen of intestine filled with gas bubbles in broiler Hubbard chicken (A,B). Jejunum filled with thick, brownish watery exudate in cobb broiler chicken (C,D).
Figure 2
Figure 2
Heat map representation of virulence, biofilm, and antimicrobial resistance profiles of C. perfringens isolates recovered from chicken, pigeon, camels, and human consumers. CT: chicken intestine, CM: chicken meat, PT: pigeon intestine, CF: camel feces, CA: camel meat, HU: human stool, M: moderate, S: strong, W: weak, MAR: multiple antibiotic resistance. Penicillin (PEN), ampicillin (AMP), amoxicillin (AMX), ampicillin-sulbactam (SAM), clindamycin (CLI), metronidazole (MTZ), imipenem (IPM), meropenem (MEM), chloramphenicol (CHL), tetracycline (TET), cefotaxime (CTX), cefoxitin (FOX), and ceftriaxone (CRO). R: resistant, S: sensitive.
Figure 3
Figure 3
Percent inhibition of biofilm formation by various concentrations of AgNPs against Clostridium perfringens. The absorbance was measured at 620 nm for the quantification of biofilm formation. CT: chicken intestine, CM: chicken meat, PT: pigeon intestine, CF: camel feces, CA: camel meat, HU: human stool.
Figure 4
Figure 4
Dendrogram showing the relatedness of C. perfringens isolated from different sources as determined by RAPD-PCR fingerprinting using the SPSS computer software program. (CT: chicken intestine, CM: chicken meat, PT: pigeon intestine, CF: camel feces, CA: camel meat, HU: human stool).
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