Hematological investigations in a case of intravascular hemolysis due to Clostridium perfringens infection.

A patient presented with fever, severe pain and edematous tight due to hip trauma and was scheduled for urgent fasciotomy. Following physical examination, laboratory analyses were requested, and results revealed anemia and severe infection. As the patient's condition was serious, a new set of samples was sent to the laboratory four hours later. Following centrifugation, severely hemolyzed dark-colored serum and plasma samples were obtained and in vitro hemolysis was suspected. The collection of samples was repeated, but a new set of samples was also hemolyzed with a significant decrease in the hemoglobin value. At that point, in vivo hemolysis was suspected, and samples were processed according to standard laboratory procedures for hemolytic samples. Following confirmation of the gas gangrene diagnosis by clinicians, the cause of hemolysis was attributed to the cytotoxic activity of α-toxin produced by the anaerobic gram-positive bacterium Clostridium perfringens. An insight into the laboratory procedure that could help to narrow down the causes of hemolysis and single out C. perfringens as a cause of intravascular hemolysis was given.

Clostridium perfringens phospholipase C, an archetypal bacterial virulence factor, induces the formation of extracellular traps by human neutrophils.

Neutrophil extracellular traps (NETs) are networks of DNA and various microbicidal proteins released to kill invading microorganisms and prevent their dissemination. However, a NETs excess is detrimental to the host and involved in the pathogenesis of various inflammatory and immunothrombotic diseases. Clostridium perfringens is a widely distributed pathogen associated with several animal and human diseases, that produces many exotoxins, including the phospholipase C (CpPLC), the main virulence factor in gas gangrene. During this disease, CpPLC generates the formation of neutrophil/platelet aggregates within the vasculature, favoring an anaerobic environment for C. perfringens growth. This work demonstrates that CpPLC induces NETosis in human neutrophils. Antibodies against CpPLC completely abrogate the NETosis-inducing activity of recombinant CpPLC and C. perfringens secretome. CpPLC induces suicidal NETosis through a mechanism that requires calcium release from inositol trisphosphate receptor (IP3) sensitive stores, activation of protein kinase C (PKC), and the mitogen-activated protein kinase/extracellular signal-regulated kinase (MEK/ERK) pathways, as well as the production of reactive oxygen species (ROS) by the metabolism of arachidonic acid. Proteomic analysis of the C. perfringens secretome identified 40 proteins, including a DNAse and two 5´-nucleotidases homologous to virulence factors that could be relevant in evading NETs. We suggested that in gas gangrene this pathogen benefits from having access to the metabolic resources of the tissue injured by a dysregulated intravascular NETosis and then escapes and spreads to deeper tissues. Understanding the role of NETs in gas gangrene could help develop novel therapeutic strategies to reduce mortality, improve muscle regeneration, and prevent deleterious patient outcomes.

Autolysin as a fibronectin receptor on the cell surface of Clostridium perfringens.

OBJECTIVE: Clostridium perfringens causes food poisoning and gas gangrene, a serious wound-associated infection. C. perfringens cells adhere to collagen via fibronectin (Fn). We investigated whether the peptidoglycan hydrolase of C. perfringens, i.e., autolysin (Acp), is implicated in Fn binding to C. perfringens cells. METHODS: This study used recombinant Acp fragments, human Fn and knockout mutants (C. perfringens 13 acp::erm and HN13 ΔfbpC ΔfbpD). Ligand blotting, Western blotting analysis, and complementation tests were performed. The Fn-binding activity of each mutant was evaluated by ELISA. RESULTS: From an Fn-binding assay using recombinant Acp fragments, Fn was found to bind to the catalytic domain of Acp. In mutant cells lacking Acp, Fn binding was significantly decreased, but was restored by the complementation of the acp gene. There are three known kinds of Fn-binding proteins in C. perfringens: FbpC, FbpD, and glyceraldehyde-3-phosphate dehydrogenase. We found no difference in Fn-binding activity between the mutant cells lacking both FbpC and FbpD (SAK3 cells) and the wild-type cells, indicating that these Fn-binding proteins are not involved in Fn binding to C. perfringens cells. CONCLUSIONS: We found that the Acp is an Fn-binding protein that acts as an Fn receptor on the surface of C. perfringens cells.

A 52-Year-Old Diabetic Man with Poor Dental Hygiene, Right Dental Abscess, and Parapharyngeal Abscess with Gas Gangrene Due to Klebsiella pneumoniae Infection, Presenting with Septic Shock and Multiorgan Failure, Who Recovered After Surgical Neck Debridement.

BACKGROUND Gas gangrene is a rapidly progressive and potentially fatal infection that is common in the extremities. Gas gangrene of the head and neck is a very rare condition, and non-clostridial gas-forming neck abscess caused by Klebsiella pneumoniae is unusual. This report is of a diabetic man with poor dental hygiene, a right dental abscess, and parapharyngeal abscess with gas gangrene due to Klebsiella pneumoniae infection, presenting with septic shock and multiorgan failure, who recovered after surgical neck debridement. CASE REPORT A 52-year-old man with diabetes mellitus lost consciousness and collapsed on a curbside. He presented with painful swelling of the right-side neck, associated with spiking fever, confusion, dyspnea, and stridor. He had right submandibular and supraclavicular swelling with crepitus, multiple dental caries, and multiorgan dysfunction, and was intubated. A computed tomography scan showed a gas-forming abscess in the right parapharyngeal, retropharyngeal, and paralaryngeal spaces and dense infiltration with pleural effusion in the upper lobes. Neck exploration was performed for drainage. Necrotic tissue and foul-smelling pus were debrided and drained. Gram stain showed gram-negative bacilli. Necrotic tissue, pus, and blood culture showed Klebsiella pneumoniae. He remained on intravenous meropenem for 14 days and was frequently debrided with irrigation until the infection subsided. Finally, normal physiologic functions of the failing organ system were restored. CONCLUSIONS We present a rare case of Klebsiella pneumoniae infection causing gas gangrene in the deep neck spaces, leading to septic shock and multiorgan failure, who recovered after surgical neck debridement. This is a potentially fatal condition that requires emergency drainage because of its high mortality rate.

Phagocyte extracellular traps formation contributes to host defense against Clostridium perfringens infection.

Clostridium perfringens (C. perfringens) is an important Gram-positive anaerobic spore-forming pathogen that provokes life-threatening gas gangrene and acute enterotoxaemia, although it colonizes as a component of the symbiotic bacteria in humans and animals. However, the mechanisms by which C. perfringens is cleared from the host remains poorly understood, thereby impeding the development of novel strategies for control this infection. Here, we uncover a beneficial effect of extracellular traps (ETs) formation on bacterial killing and clearance by phagocytes. C. perfringens strain ATCC13124, and wild-type isolates CP1 and CP3 markedly trigger ETs formation in macrophages and neutrophils. As expected, visualization of DNA decorated with histone, myeloperoxidase (MPO) and neutrophils elastase (NE) in C. perfringens-triggered classical ETs structures. Notably, the bacteria-induced ETs formation is an ERK1/2-, P38 MAPK-, store-operated calcium entry (SOCE)-, NADPH oxidase-, histone-, NE-, and MPO-dependent process, and is independent of LDH activity. Meanwhile, the defect of bactericidal activity is mediated by impairing ETs formation in phagocytes. Moreover, In vivo studies indicated that degradation of ETs by DNase I administration leads to a defect in the protection against experimental gas gangrene, with higher mortality rates, exacerbated tissue damage, and more bacterial colonization. Together, these results suggest that phagocyte ETs formation is essential for the host defense against C. perfringens infection.

Postpartum clostridial gangrenous metritis in 12 dairy goats in France.

Clostridial infections in goats have been associated frequently with enteric diseases or gas gangrene but very rarely with the reproductive system. We describe here 12 cases of fatal postpartum gangrenous metritis in does associated with infection by several clostridial species. Clinically, these cases were characterized by rapid onset of hyperthermia followed by death after kidding. On postmortem examination, the uteri appeared to be necrotic and were hemorrhagic and edematous. Microscopically, the uteri had diffuse coagulative necrosis, edema, hemorrhage, and fibrinous thrombi with intralesional gram-positive rods. Clostridium perfringens was isolated from 7 of 9 uterine samples cultured, and C. perfringens, C. septicum, C. novyi, or C. chauvoei were demonstrated by immunohistochemistry (IHC) in the 5 cases examined. IHC for Paeniclostridium sordellii was negative in all 5 cases. PCR performed on 3 of the C. perfringens isolates was positive for alpha toxin and perfringolysin, identifying these isolates as type A. Clostridial infection should be considered in cases of postpartum gangrenous metritis of does.

Gas-Producing Infections in the Foot at a Large Academic Medical Center: A 10-Year Retrospective Review.

Gas-producing infections, such as clostridial and nonclostridial gas gangrene, crepitant cellulitis, and necrotizing fasciitis, are characterized in the literature by a variety of initial presentations, microbial burdens and surgical outcomes-ranging from debridement to amputation to death. The primary aim of this study was to identify the organisms cultured in gas-producing infections of the foot in patients that presented to a large academic medical center over a 10-year period. Our secondary aims were to report the prevalence of sepsis in this population upon presentation, and patient outcomes upon discharge. After a retrospective chart review of 207,534 procedures, 70 surgical cases met inclusion criteria. The most common organisms that grew in operating room cultures were Staphylococcus aureus, Group B Beta Streptococcus, and Enterococcus species. Just over half of the population presented with sepsis. After an average of 2 or more operations, 64% of patients underwent amputation. One death occurred. Gas-producing infections, or "gas gangrene," are primarily polymicrobial infections, rarely due to Clostridium perfringens, that warrant surgical exploration for optimal outcomes.

Transtentorial herniation syndrome from meningococcal meningitis in a young woman: the case for neurocritical care.

We report a case of a previously healthy early adolescent female who presented with meningococcal meningitis. While in hospital, she had marked neurologic deterioration with clinical herniation from malignant cerebral oedema. She was transferred to a neurocritical care centre where she underwent invasive intracranial pressure (ICP) and brain tissue oxygen (PbtO2) monitoring. Early in her course, she demonstrated a compete absence of autoregulation, with pressure passive cerebral blood flow. As a result, maintaining a mean arterial pressure between 50 mm Hg and 60 mm Hg, which ensured adequate cerebral oxygenation, while avoiding increases in ICP. Although her course was initially complicated by bilateral optic neuropathy, she has subsequently made a full neurologic recovery and is now undertaking postsecondary education. This case highlights that access to specialist neurocritical care, guided by neurophysiologic monitoring of ICP and PbtO2, may help improve outcomes, even among those patients with catastrophic cerebral oedema from bacterial meningitis.

Piceatannol Alleviates Clostridium perfringens Virulence by Inhibiting Perfringolysin O.

Clostridium perfringens (C. perfringens) is an important foodborne pathogen that can cause diseases such as gas gangrene and necrotizing enteritis in a variety of economic animals, seriously affecting public health and the economic benefits and healthy development of the livestock and poultry breeding industry. Perfringolysin O (PFO) is an important virulence factor of C. perfringens and plays critical roles in necrotic enteritis and gas gangrene, rendering it an ideal target for developing new drugs against infections caused by this pathogen. In this study, based on biological activity inhibition assays, oligomerization tests and computational biology assays, we found that the foodborne natural component piceatannol reduced pore-forming activity with an inhibitory ratio of 83.84% in the concentration of 16 µg/mL (IC50 = 7.83 µg/mL) by binding with PFO directly and changing some of its secondary structures, including 3-Helix, A-helix, bend, and in turn, ultimately affecting oligomer formation. Furthermore, we confirmed that piceatannol protected human intestinal epithelial cells from the damage induced by PFO with LDH release reduced by 38.44% at 16 µg/mL, based on a cytotoxicity test. By performing an animal experiment, we found the C. perfringens clones showed an approximate 10-fold reduction in infected mice. These results suggest that piceatannol may be a candidate for anti-C. perfringens drug development.

Infectious shock after liposuction.

BACKGROUND: Liposuction 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 PRESENTATION: A 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. CONCLUSIONS: Failure 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.

Clostridial panophthalmitis - An unusual case of localised malignant oedema in Merino lambs associated with Clostridium chauvoei.

The authors report an unusual case of Clostridium chauvoei causing severe panophthalmitis in Merino lambs. More than half of the lambs affected survived, which is unusual for clostridial disease; however, there have been reports in humans that the mortality risk for ocular gas gangrene is lower than when other body parts are affected. A combination of factors in this case included environmental contamination (specific to a particular paddock), genetics predisposing to entropion (lambs born of maiden 2-year-old ewes with some inbreeding), the practice of manually everting eyelids for the entropion and vaccination strategies. C. chauvoei was cultured in pure growth from the eye of a lamb affected by severe panophthalmitis. Histopathology was consistent with severe acute infection, and microscopic sections showed Gram-positive organisms associated with the inflammatory response in the eye. In the acutely affected animal examined there were no signs that the lesions were long-standing. Animals that recovered had one or both eyes destroyed. This report describes that malignant oedema in sheep due to C. chauvoei can manifest as acute and severe panophthalmitis. The case fatality rate of lambs with panophthalmitis was less than 50%, lower than normally occurs for clostridial diseases. Clostridial vaccination of the ewes may have provided a low level of protection in reducing the case fatality rate in the affected lambs as well localisation of the infection from the blood-ocular barrier.

[Fatal necrotizing fasciitis with clostridium perfringens infection following resection of soft tissue sarcoma of the thigh]. / Letaler Verlauf einer fulminanten nekrotisierenden Fasziitis und Clostridium-perfringens-Infektion nach Resektion eines Weichteil-Sarkoms.

We present the case of a 57-year-old male patient with a fatal outcome after resection of a soft tissue sarcoma of the lateral thigh. A polymicrobial surgical site infection with Staphylococcus lugdunensis and Clostridium perfringens caused fulminant necrotising fasciitis with an additional gas gangrene. The patient suffered a severe sepsis with consecutive haemolysis and multiorgan failure. The authors recapitulate the deadly progress of a rarely reported complication after oncological resection. The therapeutic approach and surgical interventions are discussed based on the current literature.

Fatal non-traumatic gas gangrene caused by Clostridium perfringens type A in a Siberian Husky dog.

An 8-y-old, castrated male Siberian Husky dog was admitted to an emergency clinic with acute collapse and severe swelling of both forelimbs, ventral thorax, and axillary region. The clinical assessment, with laboratory tests and radiologic investigation, confirmed severe subcutaneous emphysema and multi-organ failure. The animal died while receiving emergency treatment. On postmortem examination, Clostridium perfringens was isolated from the subcutaneous fluid and the effusion from the thoracic and abdominal cavities. Relevant histopathology findings included subcutaneous emphysema and multi-organ perivascular and intravascular, intralesional myriad 0.5-3-µm gram-positive rod bacteria, with no associated inflammation. Whole-genome sequencing and phylogenetic analysis identified C. perfringens type A. Virulence genes detected included cpa (alpha toxin), cadA (v-toxin), colA (collagenase A), nagH (hyaluronidase), nanH, nanI, nanJ (sialidases), and pfoa (perfringolysin). These virulence genes have previously been reported to act synergistically with alpha toxin in C. perfringens-mediated gas gangrene.

[Soft tissue necrotizing infection]. / L'image du mois. Infection nécrosante des tissus mous.

Necrotizing fasciitis and gas gangrene represent a rapidly progressive and fatal tissue infection in the absence of early multidisciplinary treatment. There are multiple risk factors, but diabetes remains the main one. The presence of crackles or extensive lesions are an indication for exploration and surgical treatment. Conservative management is associated with zero survival. Iconography, biology or bacteriology can help in the diagnosis, but the latter is only made during surgery. The associated mortality is high, despite appropriate management. We report here the case of a diabetic patient requiring insulin, having presented this pathology, the starting point of which is a traumatic wound, with an extensive acute course, contra-indicating any surgical procedure.

La fasciite nécrosante et la gangrène gazeuse représentent une infection tissulaire rapidement progressive et létale en l'absence d'un traitement multidisciplinaire précoce. Il existe de multiples facteurs de risques, mais le diabète en reste le principal. La présence de crépitants ou de lésions extensives sont une indication à l'exploration et au traitement chirurgical. Une prise en charge conservatrice est associée à une survie nulle. L'iconographie, la biologie ou la bactériologie permettent d'aider au diagnostic, mais ce dernier est uniquement posé en per-opératoire. La mortalité associée est élevée, malgré une prise en charge adaptée. Nous rapportons ici le cas d'un patient diabétique de type 2 insulino-requérant, ayant présenté cette pathologie dont le point de départ est une plaie traumatique, avec une évolution aiguë extensive, contre-indiquant tout geste chirurgical.