Clostridium Perfringens Infections Research Articles

Repurposing rabeprazole sodium as an anti-Clostridium perfringens drug by inhibiting perfringolysin O.

Clostridium perfringens infections affect food safety, human health, and the development of the poultry feed industry. Anti-virulence is an alternative strategy to develop new drug. Perfringolysin O (PFO) is an exotoxin of C. perfringens that has been demonstrated to play critical roles in the pathogenesis of this organism, promising it an attractive target to explore drugs to combat C. perfringens infection. Based on an activity-based screening, we identified six PFO inhibitors from the Food and Drug Administration (FDA)-approved drug library, among which rabeprazole sodium (RS) showed an optimal inhibitory effect with an IC50 of 1.82±0.746µg ml-1. The GLY57, ASP58, SER190, SER193-194, ASN199, GLU204, ASN377, THR379, and ALA200 in PFO interacted with RS during binding based on an energy analysis and H-bond analysis. This interaction blocked the oligomer formation of PFO, thereby inhibiting its cytotoxicity. RS treatment significantly increased the survival rate and alleviated pathological damage in C. perfringens or PFO-treated Galleria mellonella. RS could potentially be used as a candidate drug for treating C. perfringens infection.

Gut-targeted nanoparticles deliver specifically targeted antimicrobial peptides against Clostridium perfringens infections.

Specifically targeted antimicrobial peptides (STAMPs) are novel alternatives to antibiotics, whereas the development of STAMPs for colonic infections is hindered by limited de novo design efficiency and colonic bioavailability. In this study, we report an efficient de novo STAMP design strategy that combines a traversal design, machine learning model, and phage display technology to identify STAMPs against Clostridium perfringens. STAMPs could physically damage C. perfringens, eliminate biofilms, and self-assemble into nanoparticles to entrap pathogens. Further, a gut-targeted engineering particle vaccine (EPV) was used for STAMPs delivery. In vivo studies showed that both STAMP and EPV@STAMP effectively limited C. perfringens infections and then reduced inflammatory response. Notably, EPV@STAMP exhibited stronger protection against colonic infections than STAMPs alone. Moreover, 16S ribosomal RNA sequencing showed that both STAMPs and EPV@STAMP facilitated the recovery of disturbed gut microflora. Collectively, our work may accelerate the development of the discovery and delivery of precise antimicrobials.

Molecular epidemiology and genetic characterization of Clostridium perfringens infections in lambs

Clostridium perfringens is gram positive bacterium, wide spread in environment causing various diseases in animals and human. The current study was conducted to indentify the genetic identity of C. perfringens isolates from lambs from Egypt. Using specific primers amplifying genes associated to the toxins produced by C. perfringens, multiplex PCR was used to confirm C. perfringens in 87 out of 140 samples were collected from diseased and suspected lambs. The isolates were classified as type A in 49.4%, type B in 31.1% and type D in 19.5% of isolates. The phylogenetic analysis for the partial sequences of C. perfringens strains based on plc gene, cpb gene and etx gene obtained in the present study showed high degree of similarity with other sequences of C. perfringens strains in GenBank, isolating from sheep from Egypt and other countries. According to the findings, lambs with enterotoxaemia more frequently have C. perfringens type A and an efficient hygienic control program is necessary to reduce the infection spreading among susceptible animals.

Necrotizing soft tissue infection: clinical characteristics, diagnosis, and management of 32 cases in Beijing.

ObjectiveNecrotizing soft tissue infection (NSTI) is a rare but life-threatening bacterial infection. This study was performed to analyze the clinical characteristics of patients with NSTI admitted to the intensive care unit of a trauma and burn center in Beijing and to summarize the treatment experience.MethodsThis retrospective study involved patients with NSTI admitted to the intensive care unit from January 2010 to January 2020. The clinical manifestations, pathogens, laboratory test results, and prognosis were compared between survivors and nonsurvivors.ResultsThirty-two patients were enrolled (28 men, 4 women), including 25 (78.1%) survivors and 7 (21.9%) nonsurvivors. The patients’ median age was 41.5 years (range, 30.0–52.5 years). Mortality was significantly higher in patients with a blood urea nitrogen concentration of ≥11.5 mg/dL, creatinine concentration of <3.4 mg/dL, prothrombin time of ≥15.9 s, and international normalized ratio of ≥1.3. Streptococcus pyogenes and Clostridium perfringens infections were associated with higher mortality.ConclusionsMore attention should be given to elderly patients and those with hyponatremia, coagulation disorder, and higher blood urea nitrogen or creatinine concentrations. The finding of large gram-positive rods or large numbers of gram-positive cocci in wound secretion smears is a helpful early warning sign of highly lethal NSTI.

Effects of Eimeria maxima and Clostridium perfringens infections on cecal microbial composition and the possible correlation with body weight gain in broiler chickens

With the voluntary and regulatory withdrawal of antibiotic growth promoters from animal feed, coccidiosis and necrotic enteritis (NE) emerge as the top two enteric poultry infectious diseases responsible for major economic loss worldwide. The objective of this study was to investigate the correlation between the cecal microbiota compositions with the growth trait after coccidiosis and NE. In this study, the effects of Eimeria maxima and/or Clostridium perfringens infections on the microbial composition and potential correlation with the body weight gain were investigated in broiler chickens using 16S rRNA gene sequencing. E. maxima and C. perfringens coinfection successfully induced NE with its typical gut lesions and significant reductions in the percentage of relative body weight gain (RBWG%). The NE challenge model did not affect cecal microbial diversity, but influenced the cecal microbial composition. KEGG enzymes in microbiota were significantly altered in abundance following dual infections. Furthermore, significant correlations between cecal microbiota modules and RBWG% were identified in the sham control, E. maxima or C. perfringens infected groups. Understanding of host-microbiota interaction in NE would enhance the development of antibiotics-independent strategies to reduce the harmful effect of NE on the gut microbiota structure, and improve the gut health and poultry production.

Immunization with a subunit of housekeeping proteins provide partial protection against experimental necrotic enteritis in broiler chickens

Abstract Necrotic enteritis (NE) due to Clostridium perfringens (CP) infections is a prevalent enteric infectious disease accounting for $6 billion economical loss in global poultry industry. The increasing incidence of NE has been associated with the reduction and withdrawal of antibiotic growth promoters from animal feed during recent years. Therefore, the development of effective vaccines specific for NE assumes a priority for poultry industry. By far, no any effective vaccines are commercially available in broiler chickens. Our aim was to identify the potential CP proteins as vaccine targets for NE. We expressed 4 different recombinant CP proteins targeting 6 antigens: 3 major virulence-related toxins (alpha-toxin, NetB and TpeL), and 3 pilus subunit or enzymes (Collagen adhesion protein (Cna), Fructose-1,6-bisphosphate aldolase (FBA) and a Hypothetic protein (HP)), and their protection efficacies were evaluated with a severe Coccidiosis/NE challenge model using netB+tpeL+ CP strain. Young chicks were immunized twice subcutaneously with adjuvanted CP proteins on days 4, and 15. Prior to challenge, the chickens immunized with related antigens had much higher serum antibody titers for Cna and FBA/HP antigens than those with any of 3 toxin antigen-immunized groups. Following the challenge, the Cna-immunized group showed the least mortality and lowest lesion score among the individual antigen-vaccinated groups, while the pooled antigens-immunized group demonstrated no mortality against virulent challenges. The results indicate that the immunization with CP housekeeping protein, Cna, conferred better protection than other conventional virulence-related toxin proteins. Mucosal delivery system is being investigated next.

Isolation and Genotyping Study of Clostridium Perfringens From Broiler Farms Infected with Necrotic Enteritis in Sulaimania Province

The study is conducting to isolate and toxinotype the suspected cases of Clostridium perfringens infections of broiler farms in Sulaimania province. A total of 108 samples were collected from intestinal contents, mucosal scraping, and hemorrhagic lymphoid nodules from suspected cases of necrotic enteritis in broilers. The result of isolated and identified bacteria were revealed that 63 (58%) out of 108 samples were positive for C. perfringens. The results revealed that the isolates were only positive for alpha and beta2 toxin genes. Phylogenetic and DNA sequence analysis of cpa and cpb2 gene showed that cpa genes were highly identical to isolates from broiler in Iran, poultry stool and broiler in Brazil, and blue calves in Belgium. While cpb2 gene is closely related to the isolates of broiler in Iran, India and isolates of goat in Pakistan. The results indicated that the causative agent of necrotic enteritis in broiler farms in the region was mainly due to C. perfringens type A infection

Antimicrobials Used for the Therapy of Necrotic Enteritis and Coccidiosis in Broiler Chickens and Turkeys in Canada, Farm Surveillance Results (2013-2017).

Since 2013, the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) has collected antimicrobial use (AMU) and antimicrobial resistance data from sentinel broiler chicken flocks (Br, five provinces) and sentinel turkey flocks (Tk, one province 2013-2015, three provinces 2016-2017). The objectives of this paper were to describe various preventive strategies aimed at controlling necrotic enteritis (NE) and coccidiosis in the broiler chicken and turkey flocks participating in CIPARS and FoodNet Canada Farm Surveillance Program between 2013 and 2017, to quantify and identify trends in antimicrobials used in feed, and to describe temporal changes in the diagnoses of bacterial and protozoal diseases in relation to antimicrobial use in feed. Comprehensive data were collected (by questionnaire) enabling AMU assessment by various count-based metrics (i.e., frequency and number of medicated rations), weight-based metrics (i.e., inclusion rate in feed and kilograms consumed), and technical indicators (i.e., milligrams per population correction unit [mg/PCU]). Qualitative information such as reasons for use and frequency of diagnosed diseases provided context to the trends in AMU. Between 2013 and 2017, 646 broiler flocks (14.9 million kg biomass) and 234 turkey flocks (12.4 million kg biomass) were surveyed. Overall, antimicrobials used for the prevention of Clostridium perfringens infections (NE) contributed to 85% (109/128 mg/PCUBr) and 95% (59/62 mg/PCUTk) of the quantity of antimicrobials administered via feed in broiler chickens and turkeys, respectively. Three NE programs were used: either 1, 2, or ≥3 antimicrobials administered throughout the production cycle. The treatment protocol in which a single antimicrobial was used throughout the cycle was the most frequent NE preventive program for broiler chickens (58%) and turkeys (76%). Bacitracin and virginiamycin were the top two most frequently used antimicrobials in both species for NE. For coccidiosis control, ionophores and chemical coccidiostats contributed to 66% (3091 kg) and 68% (1561 kg) of the total feed antimicrobial exposures in broiler chickens and turkeys, respectively. Documented coccidiosis programs included continuous or straight (1 drug/cycle), shuttle or dual control (≥2 drugs/cycle), and vaccination. Variations in coccidiosis programs between species were noted: broiler chickens frequently used a shuttle or dual-control program (68%), whereas turkey flocks used primarily a continuous or straight program (74%). Flocks raised without antibiotics and organic farms (10.3% of broiler chickens and 9.8% turkey flocks) used vaccines to prevent coccidiosis. A small number of broiler flocks (n = 6) used a combination of a vaccination and a coccidiostat during the cycle. During the surveillance timeframe used for this paper, the total feed AMU decreased over time in broiler chickens from 136 to 120 mg/PCUBr and in turkeys from 85 to 62 mg/ PCUTk, with no remarkable changes in the frequency of flocks diagnosed with bacterial and protozoal diseases. Surveillance findings such as these will be used as valid reference points in light of the upcoming changes in Canadian federal AMU regulations and industry-led initiatives aimed at reducing AMU.

Interrelationships between Multiple Climatic Factors and Incidence of Foodborne Diseases.

Climatic factors can affect the incidence of foodborne diseases (FBDs). Moreover, microbial network inference is useful for predicting the interrelationships between the incidence of FBDs and climatic factors. However, the interrelationships between FBD pathogens and most climatic factors are unknown. Using principal component analysis (PCA) and partial correlation coefficient matrices (PCCMs), we determined the intra-ecosystem interrelationship network of the multiple combined effects of 5 climatic factors (temperature, relative humidity, rainfall, insolation, and cloudiness) and the monthly incidences of 12 bacterial FBDs. Many FBD pathogens are interrelated with multiple combined factors. Salmonellosis has strong positive interrelationships with Vibrio parahaemolyticus and enterohemorrhagic Escherichia coli, and the interrelationships between Staphylococcus aureus/enteropathogenic E. coli/enterotoxigenic E. coli exhibits a typical triangular pattern with the combined effects of all 5 climatic factors. Meanwhile, campylobacteriosis and Clostridium perfringens infections are negatively interrelated with insolation and cloudiness. Enteroinvasive E. coli, Bacillus cereus, Listeria spp., and Yersinia enterocolitica are significantly interrelated with any climatic factor combination. The interrelationships or higher-order interrelationships among these climatic factors play an important role in the incidence of FBDs, although the underlying mechanisms remain unclear. Our results will serve as a foundation for more sophisticated models of future FBD patterns with regard to climate change.

Clinical manifestation of Campylobacter enteritis in children.

PurposeTimely antibiotic therapy in selected cases of diarrhea associated with bacterial infections can reduce the duration and severity of illness and prevent complications. The availability of a predictive index before identification of causative bacteria would aid in the choice of a therapeutic agent.MethodsThe study included patients admitted to the pediatrics unit at Konyang University Hospital for acute inflammatory diarrhea from August 1, 2015 to July 31, 2016 who underwent multiplex polymerase chain reaction testing. Of 248 patients, 83 had positive results. The clinical symptoms and blood test results were examined in 61 patients with Campylobacter spp. (25 patients), Salmonella spp. (18 patients), and Clostridium perfringens (18 patients) infections. The mean age of the 61 patients (male:femal=31:30) was 84.0±54.8 months, and the mean hospital stay was 4.6±1.7 days.ResultsThere were no statistical differences in sex, age, clinical symptoms, or signs. Patients with Campylobacter infection were significantly older (P=0.00). C-reactive protein (CRP) levels in patients with Campylobacter infection were higher than those in the other 2 groups, at 9.6±6.1 mg/dL. The results of receiver-operating characteristic curve analysis showed that the cutoff age was ≥103.5 months (sensitivity, 72%; specificity, 86%) and the CRP cutoff level was ≥4.55 mg/dL (sensitivity, 80%; specificity, 69%).ConclusionAge (≥103.5 months) and higher CRP level (≥4.55 mg/dL) were good predictors of Campylobacter enterocolitis. If neither criterion was met, Campylobacter enterocolitis was unlikely (negative predictive value 97.2%). When both criteria were met, Campylobacter enterocolitis was highly likely.

Establishment of valnemulin susceptibility breakpoint against Clostridium perfringens in rabbits

Susceptibility breakpoints provide fundamental information for rational administration of antibiotics. The present investigation reports the first valnemulin susceptibility breakpoint (MIC<0.25 μg/mL) against Clostridium perfringens infections in rabbits based on the wild-type cutoff (COWT) and the pharmacokinetics/pharmacodynamic (PK/PD) cutoff (COPD). The established susceptibility breakpoint of valnemulin might be useful in resistance surveillance of pleuromutilins and development of clinical breakpoints.

Bacteriological and molecular studies of Clostridium perfringens infections in newly born calves.

Clostridium perfringens is considered one of the important causes of calf diarrhea. Two hundred and twenty-seven clinical samples from newly born and dead diarrheic calves were examined bacteriologically and by PCR. Bacterial culture identified C. perfringens in 168 of 227 samples. A total of 144 of these isolates were lecithinase positive, indicating C. perfringens Type A. In addition, 154 isolates were positive by alpha toxin encoding gene-PCR assay. This study showed high agreement between the results of bacteriology and multiplex PCR. The multiplex PCR typed all isolates that were typed as C. perfringens Type A through bacteriologic methods, but ten samples that were lecithinase negative were positive in the multiplex PCR. The study showed the highest occurrence of C. perfringens Type A isolations from calves during the winter and autumn compared with other seasons.

Presence and characterization of pig group A and C rotaviruses in feces of Belgian diarrheic suckling piglets

The importance of group A and C rotaviruses (RVA and RVC) in the pathogenesis of diarrhea in Belgian suckling pigs is poorly investigated, and it is not known which strains are circulating in the Belgian suckling pig population. Obtaining better insights in the occurrence of both viral species in the swine population is essential in order to develop accurate diagnostic, therapeutic and prophylactic strategies to protect suckling pigs against diarrhea in a durable manner.In the present study, viral loads of RVA and RVC were quantified in diarrhea samples of suckling piglets less than 2 weeks old, collected on 36 different Belgian farms. On 22 of 36 farms tested (61%), high viral loads of RVA (6.96–11.95 log10 copies/g feces) and/or RVC (5.40–11.63 log10 copies/g feces) were detected. Seventeen RVA isolates were genotyped for their outer capsid proteins VP7 and VP4. Four different G-genotypes (G3, G4, G5 and G9) for VP7 were found together with 4 different P-genotypes (P[6], P[7], P[13] and P[23]) for VP4, in 8 different G/P combinations. All characterized RVC strains belonged to genotype G6 (VP7), except for one strain possessing the G1 genotype. VP4 genes of Belgian RVC strains were genetically heterogeneous, but were classified in the genotype P5. Most rotavirus positive samples also contained Escherichia coli, whereas Clostridium perfringens infections were mainly detected in rotavirus negative samples.Results of the present study offer better insights in the occurrence of RVA and RVC infections in Belgian diarrheic suckling piglets. As a conclusion, routine diagnostic testing for both viral species in cases of diarrhea in suckling pigs is highly recommended. Furthermore, the present findings also offer valuable information for the development of new prophylactic measures against rotavirus. Finally, the relatedness between RVC strains from pigs and other host species is described, and their possible implications in interspecies transmission events are discussed.

Preliminary screening for toxin genes amongst stock cultures of Clostridium perfringens strains isolated from dogs and calves

The spectrum of Clostridium perfringens infections ranges from food toxinosis to myonecrosis. In the current study, whole cell protein and toxin gene types were profiled in 12 randomly selected C. perfringens veterinary stock cultures from the University of Wisconsin, Madison to determine epidemio-logical similarity, or diversity amongst strains of animal origin. Whole cell protein analysis was done by SDS-PAGE while toxin gene typing was achieved by extracting DNA by boiling, DNA concentration and purity was determined by spectrophotometer and nanodrop while separation was carried out by checking it on gel electrophoresis. Multiplex PCR was used to identify the toxigenic gene-type. C. perfringens B and C. perfringens EE with established profiles were used as control strains. Isolates typed included strains cp 296, 309, 12872 (from dogs) and 304, 305, 306, 341, 342, 10754, 12218-2, 12218-3, 12473 (from calves). All 12 strains possess the cpa gene, 4 strains have cpb2, 3 strains etx, 2 strains positive for cpe and 1 for cpb. None of the strains carries the itx gene. Two strains have only cpa gene however no strains has more than two toxin gene types, with cpa-cpb2 combination be-ing more frequent. C. perfringens 305 (etx and cpa) and 342 (cpe and cpa) shared the same protein profile but belong to different toxinotype. It is evi-dent that the cpa gene is a marker for all C. perfringens strains, and similarity in protein profile is not sine qua non for toxin gene type.

Animal models to study the pathogenesis of enterotoxigenic Clostridium perfringens infections

Rabbits, mice, rats, non-human primates, sheep and cattle have been used to study the effect of Clostridium perfringens enterotoxin (CPE). CPE produces mostly necrosis of the small intestinal epithelium along with fluid accumulation in rabbits and mice. In the latter, CPE can bind to internal organs such as the liver, which induces lethal potassium levels in blood.

Clostridium perfringens Infections Initially Attributed to Norovirus, North Carolina, 2010

We investigated an outbreak initially attributed to norovirus; however, Clostridium perfringens toxicoinfection was subsequently confirmed. C. perfringens is an underrecognized but frequently observed cause of food-borne disease outbreaks. This investigation illustrates the importance of considering epidemiologic and laboratory data together when evaluating potential etiologic agents that might require unique control measures.

Pathogenesis of Clostridium Perfringens Infections: the Case of Poultry

Pathogenesis of Clostridium Perfringens Infections: the Case of Poultry

Clostridium Perfringens type A Toxin Production in 3 Commonly Used Culture Media

In vitro toxin production is an important tool not only for diagnostic purposes but also for the study of pathogenesis of Clostridium perfringens infections. The present study was carried out to compare the level of toxin production by several strains of C. perfringens type A, isolated from the intestine of animals, when cultured in 3 different conventional culture media. Six strains of C. perfringens type A isolated from the small intestine of healthy sheep were cultured in commercial cooked meat medium (CMM), brain heart infusion (BHI), and tryptone glucose yeast (TGY). Intravenous lethality in mice and phospholipase C (PLC) activity were measured in filtered culture supernatants. Lethality of culture supernatants was highest for all isolates when grown in BHI, followed by CMM. No supernatants from any isolates grown in TGY produced lethality in mice. Phospholipase C activity was highest when the isolates were grown in BHI and CMM and significantly lower when grown in TGY.

Clostridium perfringens invasiveness is enhanced by effects of theta toxin upon PMNL structure and function: the roles of leukocytotoxicity and expression of CD11/CD18 adherence glycoprotein.

Clostridium perfringens infections are characterized by the lack of an inflammatory response at the site of infection and rapidly progressive margins of tissue necrosis. Studies presented here investigated the role of theta toxin from C. perfringens in the pathophysiology of these events. Mice passively immunized with neutralizing monoclonal antibody against theta toxin and challenged with an LD100 of log phase C. perfringens had significantly less mortality than untreated controls. Intramuscular injection of killed, washed C. perfringens in mice induced a massive time-dependent influx of polymorphonuclear leukocytes (PMNL) into tissue; injection of either viable, washed C. perfringens or killed organisms plus theta toxin dramatically attenuated PMNL influx although PMNL accumulated in adjacent vessels. The anti-inflammatory effects could not be attributed to an absence of chemoattractants since C. perfringens proteins had chemotactic factor activity, and killed bacilli generated serum-derived chemotactic factors. Scanning and transmission electron microscopy demonstrated the dramatic leukocidal effects of high doses of theta toxin on PMNL. In contrast, sublethal concentrations of theta toxin primed PMNL chemiluminescence, disrupted PMNL cytoskeletal actin polymerization/disassembly, and stimulated functional upregulation of CD11b/CD18 adherence glycoprotein. In summary, these results demonstrate that theta toxin is an important virulence factor in C. perfringens infection. In a concentration-dependent fashion, theta toxin contributes to the pathogenesis of clostridial gangrene by direct destruction of host inflammatory cells and tissues, and by promoting dysregulated PMNL/endothelial cell adhesive interactions.

Influence of Bacteria on Clostridium perfringens Infections in Young Chickens

When monoflora chickens with Lactobacillus acidophilus or Streptococcus faecalis were inoculated with Clostridium perfringens either in broth culture or resuspended in Gifu anaerobic medium broth or supernatant fluid, few or no chickens died. Approximately 50% of germ-free chickens died after inoculation of C. perfringens culture, whereas no conventional birds died after inoculation of broth culture. C. perfringens in the contents of duodenum from germ-free chickens numbered about 10(4) colony-forming units (CFU)/g after inoculation 10(8) CFU broth culture per bird, but in gnotobiotic and conventional chickens this organism decreased or was not detected. When C. perfringens was cultured in intestinal contents collected from germ-free chickens, C. perfringens proliferated but alpha toxin was not detected. These findings indicate that the pathogenicity of C. perfringens was suppressed by L. acidophilus or S. faecalis administered previously or inhibited by normal intestinal flora.