Clostridium Perfringens Infection Research Articles

A mixture of organic acids and thymol protects primary chicken intestinal epithelial cells from Clostridium perfringens infection in vitro

Necrotic enteritis causes economic losses estimated to be up to 6 billion US dollars per year. Clinical and subclinical infections in poultry are also both correlated with decreased growth and feed efficiency. Moreover, in a context of increased antibiotic resistance, feed additives with enhanced antimicrobial properties are a useful and increasingly needed strategy. In this study, the protective effects of a blend of thymol and organic acids against the effects of Clostridium perfringens type A (CP) on chicken intestinal epithelial cells were investigated and compared to bacitracin, a widely used antibiotic in poultry production. Primary chicken intestinal epithelial cells were challenged with CP for a total time of 3 h to assess the beneficial effect of 2 doses of citric acid, dodecanoic acid, and thymol-containing blend, and compare them with bacitracin. During the challenge, different parameters were recorded, such as transepithelial electrical resistance, cell viability, mRNA expression, and reactive oxygen species production. CP induced inflammation with cytokine production and loss of epithelial barrier integrity. It was also able to induce reactive oxygen species production and increase the caspase expression leading to cellular death. The high dose of the blend acted similarly to bacitracin, preventing the disruptive effects of CP and inducing also an increase in zonula occludens-1 mRNA expression. The low dose only partially prevented the disruptive effects of CP but successfully reduced the associated inflammation. This study shows that the usage of thymol combined with 2 organic acids can protect primary chicken intestinal epithelial cells from CP-induced damages creating a valid candidate to substitute or adjuvate the antibiotic treatment against necrotic enteritis.

Infectious shock after liposuction

BackgroundLiposuction has become one of the most popular cosmetic surgeries in China. However, few studies have discussed infectious shock caused by C. perfringens as one of the causes of death after liposuction.Case presentationA 24-year-old woman was brought to the emergency department (ED) of Guangzhou Chinese Overseas Hospital for treatment. The patient had undergone liposuction in her bilateral lower limbs two days prior. At the ED, the patient was unconscious, and had bilateral equal-sized (diameter, 6 mm) round pupils, no light reflex, a blood pressure (BP) of 71/33 mmHg, a heart rate of 133 bpm, and an SpO2 of 70%. She had bilateral limb swelling, extensive ecchymoses in her lower abdomen and bilateral thighs, local crepitus, blisters, weak pulses on her femoral artery and dorsalis pedis, high skin tension, and hemoglobin of 32 g/L. The patient was diagnosed with Clostridium perfringens infection, and she underwent debridement surgery and supportive treatment. But the patient’s BP could not improve. At 8:28 pm on the day of admission, the patient was declared clinically dead after the electrocardiograph showed a horizontal line and spontaneous respiration ceased.ConclusionsFailure to meet surgical disinfection and environmental standards may be the cause of infection of C. perfringens through wounds. Therefore, it is necessary to strengthen the environmental disinfection of the operating room, and standardize the sterile conditions of the operation staff and patients before and during operation. Liposuction surgery necrotizing fasciitis is a rare but fatal complications, especially if diagnosis delay, therefore it is critical for early diagnosis and treatment of gas gangrene.

Diagnostic and therapeutic strategy for Clostridium perfringens infection in postpartum dairy cows: a report of 14 cases

ABSTRACT Clostridium perfringens type A infections are economically devastating to dairy farms. In spite of low morbidity, mortality is close to 100%. Although this disease has frequently been reported in calves, it is rarely seen in adult cattle, especially postpartum dairy cows. Sudden onset of symptoms, rapidly followed by death, was successively recorded in 14 postpartum dairy cows. Veterinary treatment was ineffective. The 14 cows fell ill after parturition, either within 24 h (fast invasion) or within 2–3 days (slow invasion). All the cows died shortly after symptom onset. Comprehensive analysis of epidemiology, clinical manifestation, autopsy results, and laboratory tests confirmed the 14 cases as most acute and acute C. perfringens type A infections. After the enactment of appropriate preventive and treatment measures, in conjunction with improved feeding management, no further symptoms of infection were observed in the postpartum cows. Our results demonstrated that autopsies and laboratory tests are required for the definitive diagnosis of this disease and highlighted importance of feeding management during the early perinatal period to prevent postpartum diseases in cows. This work also provides a reference for clinical veterinary diagnosis and the treatment of this disease, so as to reduce the economic losses of dairy farms.

Extracellular trap can be trained as a memory response

ABSTRACT Extracellular trap (ET) appears as a double-edged sword for the host since it participates in host immune defense by entrapping pathogens, while excessive ET release also contributes to various diseases progression including atherosclerosis, cancer, and autoimmune disorders. A better understanding of ET formation and regulation will be beneficial for developing strategies for infection control and ET-associated disease treatment. There is some evidence indicating that prior infection can enhance extracellular killing. Neutrophils from cancer or sepsis are predisposed to generate ET. It is reasonable to suspect that ET may be trained to form as a memory response, just like cytokine memory response termed “trained immunity.” The mice were intraperitoneally injected with heat-killed Candida albicans (HK-C. albicans), 3 days later bone marrow-derived macrophages (BMDM) were isolated and challenged with Clostridium perfringens as a second stimulation. We found that HK-C. albicans priming enhanced ET formation upon Clostridium perfringens infection, accompanied by increased extracellular killing capacity. Mannan priming also enhanced ET formation. Since ETs memory was induced in chicken PBMC, ETs memory may be evolutionarily conserved. Moreover, mTOR was required for ETs memory response. Collectively, this study showed that ETs can be trained as a memory response and indicated that memory property of ETs should be considered during the understanding of recurrent infection and ET-associated disorders.

Structural Changes in Trabecular Bone, Cortical Bone and Hyaline Cartilage as Well as Disturbances in Bone Metabolism and Mineralization in an Animal Model of Secondary Osteoporosis in Clostridium perfringens Infection.

There is no information regarding whether changes in the microbiological balance of the gastrointestinal tract as a result of an infection with Clostridium perfringens influence the development of metabolic bone disorders. The experiment was carried out on male broiler chickens divided into two groups: control (n = 10) and experimental (n = 10). The experimental animals were infected with Clostridium perfringens between 17 and 20 days of age. The animals were euthanized at 42 days of age. The structural parameters of the trabecular bone, cortical bone, and hyaline cartilage as well as the mineralization of the bone were determined. The metabolism of the skeletal system was assessed by determining the levels of bone turnover markers, hormones, and minerals in the blood serum. The results confirm that the disturbed composition of the gastrointestinal microflora has an impact on the mineralization and metabolism of bone tissue, leading to the structural changes in cortical bone, trabecular bone, and hyaline cartilage. On the basis of the obtained results, it can be concluded that changes in the microenvironment of the gastrointestinal tract by infection with C. perfringens may have an impact on the earlier development of osteoporosis.

Increasing head safety in broiler chickens when using probiotic Probiocid-Ultra under infection conditions of Clostridium perfringens

To prevent bacterial infections in birds, poultry farms often use feed antibiotics. Based on this, residues are often registered in products from such birds, which is dangerous for humans. Dietary supplements help to safely replace antibiotics and produce environmentally friendly products. They have a positive effect on the body of poultry: improve intestinal and microbial balance and, consequently, increase their safety and productivity.The aim of the work was to study the ability of "Probiocid® Ultra" to inhibit the development of Clostridium perfringens. In the conditions of vivariums on the basis of SRCC of poultry farming experiment with chickens-broilers of cross "Ross-308" was put. Daily chickens were divided into 3 groups of 40 chickens at random - 2 control groups and the group, in the diet of which was introduced a complex of additional food "Probiocid® Ultra".The birds were kept in the vivarium for 38 days. The microclimate, lighting, feeding and watering front complied with the requirements of Ross-308 cross. Since the age of 15 days 1 control group and the group with complex of additional feeding were infected with Clostridum perfringens at the dose of 1 billion microbial bodies per head. As a result of the experiment it was found that the studied Probiocid-Ultra has a positive effect on the poultry organism. When introducing this complex of supplementary nutrition in the diet of broilers under condi tions of Clostridium perfringens infection, a significantly higher preservation of livestock compared with the infected control was observed. Thus, it can be concluded that the use of the complex "Probiocid® Ultra" is able to protect poultry under conditions of high pathogenic load on the body.

Pneumocephalus and pneumorrhachis caused by Clostridium perfringens infection

A 68-year-old man presented with a one-month history of fever. He was bedridden due to Parkinson's disease, and a central venous catheter was inserted in his left femoral vein for parenteral nutrition at another hospital two months previously. Computed tomography revealed air in the subarachnoid space (Fig. 1A,B), in the vertebral canal (Fig. 1C), and in the left femoral vein (Fig. 1D). Three sets of blood cultures grew Clostridium perfringens. The catheter was removed and ampicillin was intravenously administered.

Supplemental Bacillus subtilis PB6 Improves Growth Performance and Gut Health in Broilers Challenged with Clostridium perfringens.

Clostridium perfringens (CP) is the principal pathogenic bacterium of chicken necrotic enteritis (NE), which causes substantial economic losses in poultry worldwide. Although probiotics are known to provide multiple benefits, little is known about the potential effects of Bacillus subtilis (B. subtilis) application in preventing CP-induced necrotic enteritis. In this study, 450 male Arbor Acres broilers were divided into 5 experimental treatments: A: basal diet (control group); B: basal diet and CP challenge (model group); C: CP challenge+10 mg/kg enramycin (positive control group); D: CP challenge+4 × 107 CFU/kg of feed B. subtilis PB6 (PB6 low-dosage group); and E: CP challenge+6 × 107 CFU/kg of feed B. subtilis PB6 (PB6 high-dosage group). There were 6 replicate pens per treatment with 15 broilers per pen. The present research examined the effect of Bacillus subtilis PB6 (B. subtilis PB6) on growth performance, mRNA expression of intestinal cytokines and tight junctions, and gut flora composition in broilers challenged with CP. The entire experiment was divided into two phases: the non-CP challenge phase (d0–18) and the CP challenge phase (d18–26). PB6 did not increase the growth performance during the first stage, but the PB6 high-dosage group was found to have larger body weight gain and ADFI during the CP challenge stage. Feed supplementation with PB6 reduced the lesion score of challenged chicks, with increased tight junction-related gene expression (occludin and ZO-1) and decreased TNF-α expression compared with CP-infected birds. A decrease in the abundance of Clostridium XI, Streptococcus, and Staphylococcus was observed after CP infection (P < 0.05), while supplementation with PB6 restored the ileal microbial composition. In conclusion, administration of B. subtilis PB6 improved growth performance, enhanced intestinal barrier function, and mitigated intestinal inflammation/lesions, which might be due to its restoring effects on the ileal microbial composition in CP-challenged broilers.

Protective Effects of Bacillus amyloliquefaciens 40 Against Clostridium perfringens Infection in Mice.

This study aimed to investigate the protective effects of Bacillus amyloliquefaciens (BA40) against Clostridium perfringens (C. perfringens) infection in mice. Bacillus subtilis PB6 was utilized as a positive control to compare the protective effects of BA40. In general, a total of 24 5-week-old male C57BL/6 mice were randomly divided into four groups, with six mice each. The BA40 and PB6 groups were orally dosed with resuspension bacteria (1 × 109 CFU/ml) once a day, from day 1 to 13, respectively. In the control and infected groups, the mice were orally pre-treated with phosphate-buffered saline (PBS) (200 μl/day). The mice in the infected groups, PB6 + infected group and BA40 + infected group, were orally challenged with C. perfringens type A (1 × 109 CFU/ml) on day 11, whereas the control group was orally dosed with PBS (200 μl/day). The results showed that the BA40 group ameliorated intestinal structure damage caused by the C. perfringens infection. Furthermore, the inflammatory responses detected in the infected groups which include the concentrations of IL-1β, TNF-α, IL-6, and immunoglobulin G (IgG) in the serum and secretory immunoglobulin (SigA) in the colon, and nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) activity in the jejunum, were also alleviated (P < 0.05) by BA40 treatment. Similarly, cytokines were also detected by quantitative PCR (qPCR) in the messenger RNA (mRNA) levels, and the results were consistent with the enzyme-linked immunosorbent assay (ELISA) kits. Additionally, in the infected group, the mRNA expression of Bax and p53 was increasing and the Bcl-2 expression was decreasing, which was reversed by BA40 and PB6 treatment (P < 0.05). Moreover, the intestinal microbiota imbalance induced by the C. perfringens infection was restored by the BA40 pre-treatment, especially by improving the relative abundance of Verrucomicrobiota (P < 0.05) and decreasing the relative abundance of Bacteroidetes (P < 0.05) in the phyla level, and the infected group increased the relative abundance of some pathogens, such as Bacteroides and Staphylococcus (P < 0.05) in the genus level. The gut microbiota alterations in the BA40 group also influenced the metabolic pathways, and the results were also compared. The purine metabolism, 2-oxocarboxylic acid metabolism, and starch and sucrose metabolism were significantly changed (P < 0.05). In conclusion, our results demonstrated that BA40 can effectively protect mice from C. perfringens infection.

Coinfection of Clostridium perfringens and Escherichia coli in gas-producing perianal abscess diagnosed by 16S rDNA sequencing: a case report

BackgroundGas-producing perianal abscess raises the possibility of clostridial infection, with Clostridium perfringens being the most common causative agent, which is highly lethal if untreated timely. As the treatment of clostridial infections often differs from that of non-clostridial infections, which they may closely resemble, the importance of accurate pathogenic organism identification cannot be overemphasized. The 16S rDNA of bacteria is highly conserved within a species and among species of the same genus but demonstrates substantial variation between different species, thus making it a suitable genomic candidate for bacterial detection and identification.Case presentationHere, we report the case of a 53-year-old patient who was admitted to the hospital for a gas-producing perianal abscess. The patient was managed with ceftizoxime and ornidazole and then received debridement and drainage at the lesion on the second day after admission. The bacterial cultures of the patient isolates from the debridement showed a coinfection of Escherichia coli and Enterococcus faecium. Although perianal redness and swelling subsided obviously after the surgery, the patient was febrile to 38.3℃ with his left upper thigh red and swollen, aggravated with tenderness and crepitus. Considering insufficient debridement and the risk of incorrect identification of pathogens, a second debridement and drainage were performed 4 days after the primary operation, and 16S rDNA sequencing of the isolates implicated Clostridium perfringens infection. Given the discrepancies in diagnostic results and the treatment outcomes, Enterococcus faecium was identified as sample contamination, and a diagnosis of coinfection of Clostridium perfringens and Escherichia coli in gas-producing perianal abscess was confirmed. The patient was then successfully treated with meropenem and vancomycin and was discharged at 27 days of admission.ConclusionsThis case represents the first report of coinfection of both clostridial and non-clostridial organisms in gas-producing perianal abscess and the first case reporting the use of 16S rDNA sequencing in the diagnosis of perianal abscess. Timely pathogen identification is critical for treating gas-producing perianal abscess and an antibiotic regimen covering both aerobic and anaerobic organisms is recommended before true pathogens are identified.

Pathogenic Characterization of Clostridium perfringens Strains Isolated From Patients With Massive Intravascular Hemolysis.

Sepsis caused by Clostridium perfringens infection is rare but often fatal. The most serious complication leading to poor prognosis is massive intravascular hemolysis (MIH). However, the molecular mechanism underlying this fulminant form of hemolysis is unclear. In the present study, we employed 11 clinical strains isolated from patients with C. perfringens septicemia and subdivided these isolates into groups H and NH: septicemia with (n = 5) or without (n = 6) MIH, respectively. To elucidate the major pathogenic factors of MIH, biological features were compared between these groups. The isolates of two groups did not differ in growth rate, virulence-related gene expression, or phospholipase C (CPA) production. Erythrocyte hemolysis was predominantly observed in culture supernatants of the strains in group H, and the human erythrocyte hemolysis rate was significantly correlated with perfringolysin O (PFO) production. Correlations were also found among PFO production, human peripheral blood mononuclear cell (PBMC) cytotoxicity, and production of interleukin-6 (IL-6) and interleukin-8 (IL-8) by human PBMCs. Analysis of proinflammatory cytokines showed that PFO induced tumor necrosis factor-α (TNF-α), IL-5, IL-6, and IL-8 production more strongly than did CPA. PFO exerted potent cytotoxic and proinflammatory cytokine induction effects on human blood cells. PFO may be a major virulence factor of sepsis with MIH, and potent proinflammatory cytokine production induced by PFO may influence the rapid progression of this fatal disease caused by C. perfringens.

Dietary Lactobacillus fermentum and Bacillus coagulans Supplementation Modulates Intestinal Immunity and Microbiota of Broiler Chickens Challenged by Clostridium perfringens.

Preventative effects of Lactobacillus fermentum and Bacillus coagulans against Clostridium perfringens infection in broilers have been well-demonstrated. The present study was conducted to investigate the modulation of these two probiotics on intestinal immunity and microbiota of C. perfringens-challenged birds. The 336 one-day-old broilers were assigned to four groups with six replicates in each group. Birds in the control were unchallenged and fed a basal diet, and birds in the three challenged groups were dietary supplemented with nothing (Cp group), 1 × 109 CFU/kg of L. fermentum (Lf_Cp group), or 1 × 1010 CFU/kg of B. coagulans (Bc_Cp group). Challenge was performed from days 14 to 20, and samples were collected on days 21 and 28. Challenge upregulated interleukin (IL)-1β and transforming growth factor (TGF)-β4 mRNA expression in jejunum on day 21, which was downregulated by B. coagulans and L. fermentum, respectively (P < 0.05). Both probiotic groups upregulated jejunal IL-1β, interferon (IFN)-γ, IL-17, and TGF-β4 on day 28 as well as IFN-γ on day 21 (P < 0.05). The Bc_Cp group increased CD3+ T cell counts in the jejunal crypt on day 21 (P < 0.05). Challenge decreased the ileal ACE index on day 21 and cecal microbial richness on day 28, which were increased by probiotic treatments, and ileal bacterial richness decreased in the Bc_Cp group on day 28 (P < 0.05). Only ileal microbiota on day 21 was distinctly affected with an R-value at 0.3116 by ANOSIM analysis (P < 0.05). Compared with the control, ileal Firmicutes increased on day 21, and ileal Bacteroidetes and cecal Proteobacteria decreased on day 28 in challenged groups (P < 0.05). Challenge increased Romboutsia spp. in the ileum as well as unclassified f_Lachnospiraceae and Ruminococcus_torques group in the cecum, and decreased Lactobacillus spp. in the ileum on day 21, which were all conversely modulated by L. fermentum (P < 0.05). Challenge increased amino acid metabolism of ileal microbiota and membrane transport of cecal microbiota, and decreased amino acid metabolism of cecal microbiota on day 21, which were conversely regulated by both probiotics (P < 0.05). In conclusion, L. fermentum and B. coagulans attenuated the intestinal inflammation and microbial dysbiosis soon after C. perfringens challenge.

Protective Effects of Lactobacillus plantarum 16 and Paenibacillus polymyxa 10 Against Clostridium perfringens Infection in Broilers.

This study aimed to investigate the protective effects of Lactobacillus plantarum 16 (Lac16) and Paenibacillus polymyxa 10 (BSC10) against Clostridium perfringens (Cp) infection in broilers. A total of 720 one-day-old chicks were randomly divided into four groups. The control and Cp group were only fed a basal diet, while the two treatment groups received basal diets supplemented with Lac16 (1 × 108 cfu·kg−1) and BSC10 (1 × 108 cfu·kg−1) for 21 days, respectively. On day 1 and days 14 to 20, birds except those in the control group were challenged with 1 × 108 cfu C. perfringens type A strain once a day. The results showed that both Lac16 and BSC10 could ameliorate intestinal structure damage caused by C. perfringens infection. C. perfringens infection induced apoptosis by increasing the expression of Bax and p53 and decreasing Bcl-2 expression and inflammation evidence by higher levels of IFN-γ, IL-6, IL-1β, iNOS, and IL-10 in the ileum mucosa, and NO production in jejunal mucosa, which was reversed by Lac16 and BSC10 treatment except for IL-1β (P < 0.05). Besides, the two probiotics restored the intestinal microbiota imbalance induced by C. perfringens infection, characterized by the reduced Firmicutes and Proteobacteria and the increased Bacteroidetes at the phyla level and decreased Bacteroides fragilis and Gallibacterium anatis at the genus level. The two probiotics also reversed metabolic pathways of the microbiota in C. perfringens-infected broilers, including B-vitamin biosynthesis, peptidoglycan biosynthesis, and pyruvate fermentation to acetate and lactate II pathway. In conclusion, Lac16 and BSC10 can effectively protect broilers against C. perfringens infection through improved composition and metabolic pathways of the intestinal microbiota, intestinal structure, inflammation, and anti-apoptosis.

Primary Emphysematous Pancreatitis Caused by Clostridium Perfringens Infection

A 66-year-old male was admitted to a close hospital for acute pancreatitis. 48h hours after developed clinical worsening. Therefore, a CT scan was performed showing signs of Emphysematous Severe Acute Pancreatitis. A transfer to our hospital was granted, being admitted to the ICU. A follow-up CT scan was performed, showing an increase in the size of abdominal collections and pneumoretroperitoneum. For this reason, an open pancreatic necrosectomy was performed. However, the patient showed an overall excellent response allowing for weaning and extubation. 15 days after the surgery, bleeding from the abdominal wall is detected. A CT scan was performed showing epigastric artery bleeding. After infructuous endovascular embolization, a surgery was required. During the stay in the ICU the patient suffered through diverse nosocomial infections by multiresistant bacteria. The patient’s condition worsened. Due to an absence in response to therapy, the patient died 58 days after being admitted to the hospital. Discussion: Clostridium’s perfringens necrotizing acute pancreatitis is a rare condition that can usually be suspected by the presence of gas around the pancreas and characteristically pneumoretroperitoneum. Its origin is usually linked to invasive procedures of the bile duct. It can also appear on collections or necrotic tissue related to previous acute pancreatitis or chronic pancreatitis. Conclusion: The Clostridium perfringens confers a raise in morbidity and mortality in spite of optimal treatment decisions. Therefore, an early diagnosis and admission to ICUs, as well as an early appliance of surgical measures, might lower the chances of adverse outcomes.

Inflammatory responses, energy metabolism enzymes, oxidative status in Clostridium perfringens infection in broilers

The aim of this study was to determine whether infection by Clostridium perfringens negatively interferes with the oxidant/antioxidant status and activity of energy metabolism enzymes (creatine kinase (CK), adenylate kinase (AK) and pyruvate kinase (PK)), as well as the zootechnical performance of broilers. A completely randomized design with three treatments, with five replications by treatment, and 10 birds by repetitions was used: T1: non-infected group; T2: group infected with C. perfringens; T3: group infected with C. perfringens, and basal diet with performance enhancers (antibiotic and coccidiostatic). At 21 days of age, the birds were experimentally infected orally with 4.0 x 108 CFU/mL C. perfringens. At strategic moments during the experimental period, zootechnical data and blood samples were collected. There were no significant differences among treatments in terms of weight gain, feed intake, or feed conversion. At 20 days, heterophils counts were significantly lower in the T3 group birds than in the others. A significant reduction in the number of total leukocytes, because of the lower number of lymphocytes, was observed in groups T2 and T3 on day 34. Neutrophil and monocyte counts were significantly lower in group T3 than in group T1 on day 34. On day 42, there were significantly lower levels of total protein and globulin in birds in T3 than in T1. On day 27, PK activity was significantly higher in groups T2 and T3; similar behavior observed on day 34 when CK and PK activities in T2 and T3 were significantly higher than those of T1. AK activity was significantly higher in the serum of T3 birds than in those of the other groups at 34 days. At 41 days, CK and PK activities were significantly higher only in birds in group T3 compared to other groups. The levels of reactive oxygen species and lipoperoxidation, as well as the activity of glutathione S-transferase, were significantly lower in birds from group T3 compared to other groups at age 20 days. After a challenge with C. perfringens, there were fluctuations in the behavior of oxidative stress biomarkers; in particular, at 41 days of age, birds in T2 had elevated levels of lipoperoxidation, different from what was seen in birds in T3 that consumed feed with a performance enhancer. Preliminary results suggest that clostridiosis affects serum activity of enzymes in the phosphotransfer network, requiring compensatory enzymatic changes in an attempt to maintain energy homeostasis. In addition, the infection reduces the number of inflammatory cells and causes oxidant/antioxidant imbalance that may contribute to the pathophysiology of the disease. Además, la infección reduce el número de células inflamatorias y provoca un desequilibrio en el estado oxidante / antioxidante que puede contribuir a la fisiopatología de la enfermedad.

Evaluation of the efficacy of chlorogenic acid in reducing small intestine injury, oxidative stress, and inflammation in chickens challenged with Clostridium perfringens type A

The goal of the study was testing the effects of chlorogenic acid (CA) supplementation on small intestine healthiness, growth performance, oxidative stress, inflammatory response, and blood biochemical indices in specific-pathogen-free (SPF) chickens after infection with Clostridium perfringens (CP) type A. In this study, 324 1-day-old male SPF chickens were randomly distributed into 6 groups: control group; CA group; CP infection group; CA + CP group; antibiotic group; antibiotic + CP group. All 1-day-old chickens were fed with CA or antibiotic in corresponding treatment group for 13 d. On the 14 d, the chickens in corresponding infection group were challenged with CP type A for 3 d. Samples in each group were collected when the chickens were 17 and 21 d old. This study proves for the first time that CA, a Chinese herbal medicine, can effectively improve growth performance, inhibit small intestine structural damage, improve antioxidant capacity, inhibit damage to ileal mucosal layer construction and tight junctions, inhibit inflammatory cytokines, and ameliorate blood biochemical indices. Therefore, this study provides data for CA being able to effectively alleviate small intestine damage caused by CP type A infection in chickens.

Saccharomyces boulardii attenuates inflammatory response induced by Clostridium perfringens via TLR4/TLR15-MyD8 pathway in HD11 avian macrophages

Macrophages are professional phagocytic cells that play a critical role in initiating immune responses by presenting antigen and phagocytic clearance. The macrophages can be targeted for immunomodulation by beneficial microbes, such as probiotics. The aim of this study is to investigate the protective effect of Saccharomyces boulardii against Clostridium perfringens infection in avian macrophage cell line HD11. In this study, HD11 macrophages were prestimulated with S. boulardii for 6 h and then infected with C. perfringens for 3 h. Results showed that S. boulardii enhanced phagocytosis and bactericidal capacity against C. perfringens by HD11 cells. The S. boulardii effectively promoted the mRNA expression of CD80, CD83, and CD197 cell-surface molecules in C. perfringens-infected HD11 cells. Moreover, we found that prestimulation with S. boulardii reduced the mRNA expression of CD40, toll-like receptor [TLR] 4, and TLR15 induced by C. perfringens and thereby downregulated the mRNA expression of myeloid differentiation primary response 88, TNF receptor associated factor 6, nuclear factor kappa-B p65 subunit, and c-Jun N-terminal kinase genes in HD11 cells. The upregulation of cytokines (interleukin [IL]-6, tumor necrosis factor alpha, and IL-10) and inducible nitric oxide synthase mRNA expression in C. perfringens-infected HD11 cells were noticeably inhibited by S. boulardii pretreatment. Conclusively, these results might provide a new insight into the role of S. boulardii in regulating avian immune defense against C. perfringens invasion and immune escape.

Bioinformatics analysis of NetF proteins for designing a multi-epitope vaccine against Clostridium perfringens infection

Clostridium perfringens is an important human and animal pathogen that is the primary causative agent of necrotizing enteritis and enterotoxemia in many types of animals. C. perfringens produces a variety of toxins, including NetF which may plays a crucial role in the pathogenesis of foal and canine necrotizing enteritis. In this study, we used several bioinformatics methods to analyze various aspects of the NetF proteins, including the physicochemical properties, secondary and tertiary structures, and the dominant B-cell and T-cell epitopes. The results showed that NetF protein was a stable and hydrophilic protein. The secondary structure of the NetF protein consisted of 2.62% alpha helixes, 6.56% beta turns, 38.69% extended strands and 52.13% random coils. Moreover, several potential B and T-cell epitopes were identified for NetF. In addition, the obtained findings from antigenicity and allergenicity evaluation remarked that this protein is immunogenic and non-allergen.Based on the results of Ramachandran plot, 94.22%, 5. 42%, and 0.36% of amino acid residues were incorporated in the favored, allowed, and outlier regions, respectively. This study provides a foundation for further investigations, and laid a theoretical basis for the development of an appropriate vaccine against C. perfringens infection.

The impact of exogenous dietary nucleotides in ameliorating Clostridium perfringens infection and improving intestinal barriers gene expression in broiler chicken

The current study aimed to evaluate the efficiency of dietary nucleotides-supplementation on broiler chickens to alleviate the intestinal Clostridium perfringens (C. perfringens) levels and its adverse effect on gut and growth performance parameters. In this study, a total of 270 one-day-old mixed broiler chicks (Cobb 500) were randomly divided into six treatment groups with three replicates of 15 chicks/ replicate. Treatment 1 (CX), a negative control group was fed corn-soybean basal diet without added nucleotides. Treatment 2 (CN 0.05) and treatment 3 (CN 0.1), consisted of chicks were fed the basal diet with the addition of nucleotides on top at two levels (0.05 and 0.1%) respectively. Treatment 4 (PX), treatment 5 (PN 0.05), and treatment 6 (PN 0.1) consisted of chicks that were challenged with C. perfringens inoculum (~4 × 108 CFU/ml) on day 14, 15, 16 and 17of the experiment and were fed diets similar to treatments 1, 2, and 3 respectively. The trial continued for 35 days. At the end of the experiment, the intestinal C. perfringens counts, microscopic lesion scores, intestinal histomorphology, intestinal barriers (occludin and mucin mRNA expression) and growth parameters were determined. The results showed that the pathogen challenge significantly (P˂0.05) increased both C. perfringens levels and intestinal lesion scores. Which adversely affects intestinal barriers and intestinal histomorphology resulting in a significant decrease (P˂0.05) in body weight gain (BWG) with an increase in feed conversion ratio (FCR). Whereas, nucleotides-supplementation, at 0.1%, significantly decreased both C. perfringens levels and intestinal lesion scores, and significantly improved intestinal barriers and intestinal histomorphology which consequently resulted in improved growth performance parameters to be nearly the same as that of the control un-supplemented group. In conclusion, nucleotides markedly ameliorated the negative effects of C. perfringens challenge by improving the intestinal barrier function and intestinal histomorphology which positively reflected on the growth performance of challenged birds.

Gastric disorders of cattle in western Rio Grande do Sul State, Brazil

ABSTRACT: A retrospective study of gastric disorders in autopsied cattle in the Western region of Rio Grande do Sul State, was performed. The exam reports of bovine necropsy of the Veterinary Pathology Laboratory, Unipampa, were analyzed in the period from 2010 to 2018. All cases in which death was primarily caused by disturbance in the gastric chambers were included. During the period evaluated, 141 cattle were necropsied. Of those, 25 had gastric disorders. Of those, 53% had alterations in the rumen, followed by abomasum (17%), involvement of two chambers (13%) and reticulum (9%). Most cases corresponded to beef cattle raised in an extensive system and most them for calf production and fattening with an average age of approximately three years. The cases occurred in farms of four different municipalities. Bullous bloat by excessive Trifolium repens ingestion was the gastric disturbance with the highest number of dead cattle observed in this study, especially in irrigated áreas of livestock farms. Cases such as lactic acidosis, ruminal alkalosis due to excessive urea ingestion and Baccharis coridifolia poisoning were also important gastric disturbances in necropsied cattle, associated especially with poor management and period of scarcity of good quality fodder. Cases of Clostridium perfringens infection were also observed in young cattle suggesting that it is an important infectious agent in the evaluated cattle herds, also showing failures in vaccination of the herds. As observed, gastric disturbances in cattle in the western region of Rio Grande do Sul have several causes. Metabolic/toxic and infectious disturbances were important causes of mortality in the herds, inducing considerable economic losses. Based on this study, it is clear that the majority of outbreaks or isolated cases occurred due to errors in the management of the properties and the vast majority of them could have been avoided with improvements in the technical qualification of the workers and simple adjustments in the farming methods. It is also emphasized the importance of the conclusive diagnosis to control these disorders, once after the orientation to the producers, was observed significant decrease in cattle losses in the farms.