Protection Efficacy of Oral Bait Probiotic Vaccine Constitutively Expressing Tetravalent Toxoids against Clostridium perfringens Exotoxins in Livestock (Rabbits)

Clostridium perfringens is an opportunistic pathogen. Its main virulence factors are exotoxins, which are the etiological agents of enteritis necroticans and enterotoxemia caused in livestock (cattle, sheep, and rabbits). Here, we demonstrated effective immune protection for rabbits against α, β, and ε exotoxins of C. perfringens provided by an oral tetravalent bait probiotic vaccine delivering α, ε, β1, and β2 toxoids of C. perfringens. Results showed that the recombinant probiotic had good segregational stability and good colonization ability in the rabbit intestinal tract. Oral administration of the probiotic vaccine can effectively elicit significant levels of antigen-specific mucosa sIgA and sera IgG antibodies with exotoxin-neutralizing activity. Additionally, oral immunization with the probiotic vaccine effectively promoted lymphoproliferation and Th1/Th2-associated cytokine production. The protection rate of immunized rabbits with the probiotic vaccine was 80% after challenging rabbits with a combination of C. perfringens (toxinotypes A, C, and D) and exotoxin mixture, which was better than the 60% provided by a commercial inactivated C. perfringens A, C, and D trivalent vaccine. Moreover, obvious histopathological changes were observed in the intestinal tissues of rabbits in the commercial vaccine and PBS groups. The bait probiotic vaccine can provide effective protection against C. perfringens exotoxins, suggesting a promising C. perfringens vaccination strategy.

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The glycoconjugate-degrading enzymes of Clostridium perfringens: Tailored catalysts for breaching the intestinal mucus barrier

Glycobiology ◽

10.1093/glycob/cwaa050 ◽

2020 ◽

Cited By ~ 3

Author(s):

Kristin E Low ◽

Steven P Smith ◽

D Wade Abbott ◽

Alisdair B Boraston

Keyword(s):

Clostridium Perfringens ◽

Bacterial Species ◽

Opportunistic Pathogen ◽

Glycoside Hydrolases ◽

Necrotic Enteritis ◽

Carbohydrate Active Enzymes ◽

Intestinal Mucus ◽

Mucosal Layer ◽

Potential Component ◽

Mucus Barrier

Abstract The gastrointestinal (GI) tract of humans and animals is lined with mucus that serves as a barrier between the gut microbiota and the epithelial layer of the intestine. As the proteins present in mucus are typically heavily glycosylated, such as the mucins, several enteric commensal and pathogenic bacterial species are well-adapted to this rich carbon source and their genomes are replete with carbohydrate-active enzymes targeted toward dismantling the glycans and proteins present in mucus. One such species is Clostridium perfringens, a Gram-positive opportunistic pathogen indigenous to the gut of humans and animals. The genome of C. perfringens encodes numerous carbohydrate-active enzymes that are predicted or known to target glycosidic linkages within or on the termini of mucus glycans. Through this enzymatic activity, the degradation of the mucosal layer by C. perfringens has been implicated in a number of GI diseases, the most severe of which is necrotic enteritis. In this review, we describe the wide array of extracellular glycoside hydrolases, and their accessory modules, that is possessed by C. perfringens, and examine the unique multimodularity of these proteins in the context of degrading the glycoconjugates in mucus as a potential component of disease.

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Oral Vaccination with Subunit Vaccines Protects Animals against Aerosol Infection with Mycobacterium tuberculosis

Infection and Immunity ◽

10.1128/iai.70.6.3111-3121.2002 ◽

2002 ◽

Vol 70 (6) ◽

pp. 3111-3121 ◽

Cited By ~ 74

Author(s):

T. Mark Doherty ◽

Anja Weinrich Olsen ◽

Laurens van Pinxteren ◽

Peter Andersen

Keyword(s):

Mycobacterium Tuberculosis ◽

Immune Responses ◽

Lipid A ◽

Vaccination Strategy ◽

Oral Immunization ◽

Subunit Vaccines ◽

Monophosphoryl Lipid A ◽

Subcutaneous Immunization ◽

Aerosol Infection

ABSTRACT Immunity against Mycobacterium tuberculosis depends largely on activation of cell-mediated responses, and gamma interferon has been shown to play a crucial role in this process in both humans and animal models. Since the lung is normally the organ in which infection is initiated and is the major site of pathology, immune responses in the lung play a significant role in restricting initial infection with M. tuberculosis. The aim of the present study was to stimulate efficient immunity in the lung by targeting the gut mucosa. Detoxified monophosphoryl lipid A (MPL) has been shown to be a relatively nontoxic adjuvant which efficiently promotes the induction of type 1 responses when it is given by the traditional subcutaneous route. We have therefore compared subcutaneous immunization of mice to oral immunization by using a model subunit vaccine carrying two immunodominant proteins from M. tuberculosis, in combination with MPL-based adjuvants. While less effective when used to prime a response, a heterologous priming and boosting vaccination strategy employing oral boosting induced significant systemic type 1 responses which equaled and surpassed those attained by subcutaneous immunization protocols. Moreover, the increased immune responses observed correlated with the induction of substantial protection against subsequent aerosol infection with virulent M. tuberculosis at levels comparable to, or better than, those obtained by multiple subcutaneous vaccinations. These results demonstrate that booster vaccinations via mucosal surfaces, by combining efficient subunit vaccines with the potent adjuvant MPL, may be an effective method of addressing some of the shortcomings of current vaccination strategies.

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Clostridium perfringens – epidemiological importance and diagnostics

Medycyna Weterynaryjna ◽

10.21521/mw.6161 ◽

2019 ◽

Vol 75 (01) ◽

pp. 6161-2019

Author(s):

NINA KOZIEŁ ◽

ELŻBIETA KUKIER ◽

KRZYSZTOF KWIATEK

Keyword(s):

Clostridium Perfringens ◽

Extracellular Enzymes ◽

Food Poisoning ◽

Immunocompromised Host ◽

Necrotic Enteritis ◽

Virulence Determinants ◽

Gastrointestinal Infections ◽

Food Borne ◽

Disease Diagnostics ◽

Enteritis Necroticans

Clostridium perfringens is one of the most widespread anaerobic spore forming bacteria found in the environment. The toxotype A of the species inhabits the gastrointestinal tract of birds and mammals exhibiting pathogenic properties in the immunocompromised host. The virulence determinants of C. perfringens are toxins and extracellular enzymes which cause gas gangrene, enteritis necroticans, food poisoning, and non-food borne gastrointestinal infections in humans. The young animals suffer from enterotoxaemia, dysentery and necrotic enteritis due to the anaerobic spore forming bacilli. This article reviews the epidemiological significance of C. perfringens and its disease diagnostics, taking into account all known to date virulence determinants of the microorganism.

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Clostridium perfringens β-toxin binding to vascular endothelial cells in a human case of enteritis necroticans

Journal of Medical Microbiology ◽

10.1099/jmm.0.008060-0 ◽

2009 ◽

Vol 58 (6) ◽

pp. 826-828 ◽

Cited By ~ 25

Author(s):

Julien Miclard ◽

Joop van Baarlen ◽

Marianne Wyder ◽

Benno Grabscheid ◽

Horst Posthaus

Keyword(s):

Endothelial Cells ◽

Clostridium Perfringens ◽

Vascular Endothelial Cells ◽

Direct Interaction ◽

Human Case ◽

Small Intestinal Mucosa ◽

Toxin Binding ◽

Intestinal Lesions ◽

Small Intestinal ◽

Enteritis Necroticans

Clostridium perfringens type C-induced enteritis necroticans is a rare but often fatal disease in humans. A consistent histopathological finding is an acute, deep necrosis of the small intestinal mucosa associated with acute vascular necrosis and massive haemorrhage in the lamina propria and submucosa. Retrospective immunohistochemical investigations of tissues from a diabetic adult who died of enteritis necroticans revealed endothelial localization of C. perfringens β-toxin in small intestinal lesions. Our results indicate that vascular necrosis might be induced by a direct interaction between C. perfringens β-toxin and endothelial cells and that targeted disruption of endothelial cells plays a role in the pathogenesis of enteritis necroticans.

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The CpAL Quorum Sensing System Regulates Production of Hemolysins CPA and PFO To Build Clostridium perfringens Biofilms

Infection and Immunity ◽

10.1128/iai.00240-15 ◽

2015 ◽

Vol 83 (6) ◽

pp. 2430-2442 ◽

Cited By ~ 18

Author(s):

Jorge E. Vidal ◽

Joshua R. Shak ◽

Adrian Canizalez-Roman

Keyword(s):

Quorum Sensing ◽

Clostridium Perfringens ◽

Biofilm Formation ◽

Extracellular Dna ◽

Wild Type ◽

Content Type ◽

Type C ◽

Enteritis Necroticans ◽

Biofilm Structure

Clostridium perfringensstrains produce severe diseases, including myonecrosis and enteritis necroticans, in humans and animals. Diseases are mediated by the production of potent toxins that often damage the site of infection, e.g., skin epithelium during myonecrosis. In planktonic cultures, the regulation of important toxins, such as CPA, CPB, and PFO, is controlled by theC. perfringensAgr-like (CpAL) quorum sensing (QS) system. Strains also encode a functional LuxS/AI-2 system. AlthoughC. perfringensstrains form biofilm-like structures, the regulation of biofilm formation is poorly understood. Therefore, our studies investigated the role of CpAL and LuxS/AI-2 QS systems and of QS-regulated factors in controlling the formation of biofilms. We first demonstrate that biofilm production by reference strains differs depending on the culture medium. Increased biomass correlated with the presence of extracellular DNA in the supernatant, which was released by lysis of a fraction of the biofilm population and planktonic cells. Whereas ΔagrBmutant strains were not able to produce biofilms, a ΔluxSmutant produced wild-type levels. The transcript levels of CpAL-regulatedcpaandpfoAgenes, but notcpb, were upregulated in biofilms compared to planktonic cultures. Accordingly, Δcpaand ΔpfoAmutants, in type A (S13) or type C (CN3685) backgrounds, were unable to produce biofilms, whereas CN3685Δcpbmade wild-type levels. Biofilm formation was restored in complemented Δcpa/cpaand ΔpfoA/pfoAstrains. Confocal microscopy studies further detected CPA partially colocalizing with eDNA on the biofilm structure. Thus, CpAL regulates biofilm formation inC. perfringensby increasing levels of certain toxins required to build biofilms.

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Enteritis Necroticans with Recurrent Enterocutaneous Fistulae Caused by Clostridium perfringens in a Child With Cyclic Neutropenia

Journal of Pediatric Gastroenterology and Nutrition ◽

10.1097/00005176-200402000-00021 ◽

2004 ◽

Vol 38 (2) ◽

pp. 213-215 ◽

Cited By ~ 8

Author(s):

Ding-You Li ◽

Ann O. Scheimann ◽

J. Glenn Songer ◽

Richard E. Person ◽

Marshall Horwitz ◽

...

Keyword(s):

Clostridium Perfringens ◽

Cyclic Neutropenia ◽

Enteritis Necroticans

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Enteritis necroticans caused by Clostridium perfringens type a

The Journal of Pediatrics ◽

10.1016/j.jpeds.2003.09.039 ◽

2004 ◽

Vol 144 (3) ◽

pp. 410 ◽

Cited By ~ 2

Author(s):

Tadashi Iwanaka ◽

Hiroshi Kawashima ◽

Hiroshi Kishimoto ◽

Mikio Kakinuma ◽

Kazuaki Arai ◽

...

Keyword(s):

Clostridium Perfringens ◽

Type A ◽

Enteritis Necroticans

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Synergistic Effects of Clostridium perfringens Enterotoxin and Beta Toxin in Rabbit Small Intestinal Loops

Infection and Immunity ◽

10.1128/iai.01848-14 ◽

2014 ◽

Vol 82 (7) ◽

pp. 2958-2970 ◽

Cited By ~ 26

Author(s):

Menglin Ma ◽

Abhijit Gurjar ◽

James R. Theoret ◽

Jorge P. Garcia ◽

Juliann Beingesser ◽

...

Keyword(s):

Clostridium Perfringens ◽

Synergistic Effects ◽

Fluid Accumulation ◽

Content Type ◽

Positive Type ◽

Intestinal Infections ◽

Type C ◽

Small Intestinal ◽

Beta Toxin ◽

Enteritis Necroticans

ABSTRACTThe ability ofClostridium perfringenstype C to cause human enteritis necroticans (EN) is attributed to beta toxin (CPB). However, many EN strains also expressC. perfringensenterotoxin (CPE), suggesting that CPE could be another contributor to EN. Supporting this possibility, lysate supernatants from modified Duncan-Strong sporulation (MDS) medium cultures of three CPE-positive type C EN strains caused enteropathogenic effects in rabbit small intestinal loops, which is significant since CPE is produced only during sporulation and sinceC. perfringenscan sporulate in the intestines. Consequently, CPE and CPB contributions to the enteropathogenic effects of MDS lysate supernatants of CPE-positive type C EN strain CN3758 were evaluated using isogeniccpbandcpenull mutants. While supernatants of wild-type CN3758 MDS lysates induced significant hemorrhagic lesions and luminal fluid accumulation, MDS lysate supernatants of thecpbandcpemutants caused neither significant damage nor fluid accumulation. This attenuation was attributable to inactivating these toxin genes since complementing thecpemutant or reversing thecpbmutation restored the enteropathogenic effects of MDS lysate supernatants. Confirming that both CPB and CPE are needed for the enteropathogenic effects of CN3758 MDS lysate supernatants, purified CPB and CPE at the same concentrations found in CN3758 MDS lysates also acted together synergistically in rabbit small intestinal loops; however, only higher doses of either purified toxin independently caused enteropathogenic effects. These findings provide the first evidence for potential synergistic toxin interactions duringC. perfringensintestinal infections and support a possible role for CPE, as well as CPB, in some EN cases.

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Complete genomic sequence and analysis of β2 toxin gene mapping of Clostridium perfringens JXJA17 isolated from piglets in China

Scientific Reports ◽

10.1038/s41598-020-79333-8 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Xiu Zeng ◽

Baosheng Liu ◽

Jiao Zhou ◽

Yimin Dai ◽

Chunsheng Han ◽

...

Keyword(s):

Clostridium Perfringens ◽

Genomic Sequence ◽

Antibiotic Resistance Genes ◽

Close Relation ◽

Illumina Miseq ◽

Opportunistic Pathogen ◽

Toxin Gene ◽

Circular Chromosome ◽

Toxin Genes ◽

Genome Homology

AbstractClostridium perfringens (Cp) is a ubiquitous opportunistic pathogen of humans and animals in the natural environment and animal intestines. The pathogenicity of Cp depends on the production of toxins encoded by genes on the chromosomes or plasmids. In contemporary literature, there is no clear consensus about the pathogenicity of CpA β2 toxin. To analyze the homology of the genome of piglet source CpA and its β2 toxin, we sequenced the whole genome of strain JXJA17 isolated from diarrhea piglets using the Illumina Miseq and Pacbio Sequel platforms. The genome was composed of a circular chromosome with 3,324,072 bp (G + C content: 28.51%) and nine plasmids. Genome and 16S rDNA homology analysis revealed a close relation of the JXJA17 strain with the JGS1495, Cp-06, Cp-16, and FORC_003 strains. These strains were isolated from different samples and belonged to different toxin-types. JXJA17 strain was found to carry two toxin genes (plc and cpb2). In contrast to other Cp strains, the cpb2 of JXJA17 was located on a large plasmid (58 kb) with no co-localization of other toxin genes or antibiotic resistance genes. Analysis of JXJA17 genome homology and its cpb2 would facilitate our further study the relationship between β2 toxin and piglet diarrhea.

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