Clinical and Laboratory Analysis of Malignant Edema (Gas Gangrene) in Cattle in Ethiopia

Clinical and Laboratory Analysis of Malignant Edema (Gas Gangrene) in Cattle in Ethiopia | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Case Report Clinical and Laboratory Analysis of Malignant Edema (Gas Gangrene) in Cattle in Ethiopia Dessalew Habte, Habtamu Addis This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6391174/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract This study documents a case of malignant edema in an adult local breed bovine in Ethiopia. The objective was to detail the clinical presentation, diagnosis, and treatment of malignant edema caused by Clostridium septicum. The affected animal, weighing 290 kg, exhibited symptoms including depression, anorexia, lameness, weakness, and swelling in the left forequarter, as well as edema in the dewlap region. The methodology involved a comprehensive clinical examination, bacterial culture, and Gram staining to confirm the presence of C. septicum. Clinical findings included an elevated respiratory rate, fever (41.7°C), and a hot, soft, and pasty edema initially localized in the knee joint, which subsequently extended to the dewlap. The affected area showed erythema and was intensely painful. Despite treatment with dexamethasone sodium phosphate, high doses of fortified procaine penicillin, multivitamin injections, and topical iodine application, the intervention proved ineffective, leading to the animal's death. This case underscores the need for further clinical and epidemiological research to enhance management and prevention strategies for malignant edema in the region. Large Animal Medicine Anaerobic infection cattle gas gangrene toxemia veterinary diagnosis Figures Figure 1 Figure 2 1. INTRODUCTION Malignant edema (ME; gas gangrene) is an acute, often fatal wound infection caused by one or more pathogenic species of Clostridium. It affects animals of all ages and grazing species, and is globally distributed, particularly in areas with soil heavily contaminated by bacterial spores. It is considered an "exogenous" infection because environmental bacterial microorganisms penetrate tissues through wounds in the skin or mucous membranes, creating an anaerobic environment (Silva et al., 2016 ; Kusiluka et al., 2016 ). The disease malignant edema is primarily caused by Clostridium septicum , an anaerobic, Gram-positive, spore-forming organism. Although C. septicum is the main cause of edematous and emphysematous conditions, other Clostridium species, such as Clostridium chauvoei, Clostridium perfringens, Clostridium sordellii, and Clostridium novyi, can also be associated with the disease, indicating mixed infections. C. septicum and other clostridia, along with their spores, are found in organic-rich, high-humidity soils and in the intestinal tracts of animals, making them ubiquitous worldwide. Infection usually occurs through the contamination of wounds caused by accidents such as castration, sterilization, shearing, unhygienic vaccination, tail docking, and parturition, which activate dormant spores in the soil (Parish et al., 1996 ; Coetzer and Tustin, 2014 ; Kapustin et al., 2020 ). The disease is typically acute, though subacute and chronic cases may occur sporadically. Clinical signs of malignant edema include anorexia, depression, weakness, and local lesions at the infection site characterized by soft, hot, and pasty swelling with marked local erythema and intense pain upon palpation. In advanced stages, the swelling becomes tense and dark, with gaseous and edematous accumulations that can be palpated as crepitus in the subcutaneous tissue around the infected area. Other signs include high fever (41 to 42°C), intoxication, muscle tremors, lameness, rigidity, and recumbency. The skin may appear tense and diffusely black or red with bruising and suffusions. In advanced stages, subnormal body temperature is common, and the affected area may become cold with the disappearance of pain due to necrosis of local nerve endings. Death often results from systemic shock and toxemia within a few hours to 2–4 days after the onset of clinical signs (Constable et al., 2017 ; D'Angelo et al., 2020; Junior et al., 2020 ). The diagnosis of malignant edema is based on characteristic clinical signs, a history of wounds, and laboratory examination using various techniques. Diagnostic confirmation can be achieved through fluorescent antibody staining of tissue smears and by observing clinicopathological changes at necropsy, such as dark red, swollen muscles filled with hemorrhagic proteinaceous exudate and minimal gas production. ME can be differentially diagnosed from other conditions, including abscesses, quarter ill, snake bites, and anthrax (Santos et al., 2019 ; Raymundo et al., 2014 ). Effective management of malignant edema involves proper wound management and the use of antibiotics such as penicillin and tetracycline. Supportive treatments, including fluid therapy, anti-inflammatory agents, and proper nutrition, are also important. Maintaining good husbandry and sanitary practices during invasive procedures; such as shearing, castration, surgical and obstetric manipulations, tail docking, and injection administration is essential for preventing malignant edema. In endemic areas, immunization with multivalent vaccines combining C. septicum and C. chauvoei , especially before procedures like dehorning, castration, or tail docking, is crucial (Abdolmohammadi and Zahmatkesh, 2021 ; Coetzer and Tustin, 2014 ). Malignant edema causes significant economic losses due to reduced productivity and reproductive capacity, as well as the costs of treatment, control, and prevention, alongside animal morbidity and mortality in Ethiopia. All livestock species, regardless of age, are at risk of infection from endemic and/or sporadic strains of this bacterial disease, primarily caused by C. septicum in cattle, with no clear framework for diagnosis, treatment, and preventive measures with available vaccines (Roger et al., 2011 ; Gazioglu et al., 2018 ). This case report describes an episode of malignant edema caused by Clostridium bacteria in an adult local breed bovine. 2. MATERIALS AND METHODS 2.1. Case Description An adult local breed bull, weighing 290 kg, was presented to the Veterinary Clinic of Debre Markos in Debre Markos town, Ethiopia, on April 4, 2023, with severe symptoms including depression, anorexia, lameness, weakness, and swelling in the left forequarter, followed by subcutaneous edema in the neck region. The swelling developed after a wound of unknown cause, which occurred approximately one week prior to the clinic visit. The physical examination revealed a respiratory rate of 32 breaths/minute, a body temperature of 41.7°C, a heart rate of 84 beats/min, and a soft, hot, and pasty swelling that indented upon palpation. Initially, the swelling was localized in the knee joint and later extended to the subcutaneous tissue of the dewlap, with marked local erythema and intense pain at the site of infection. The skin was stretched, diffusely red or black, with bruising and suffusions. The clinical examination also noted weakness, anorexia, depression, salivation, intoxication, high fever (41.7°C), lameness with a stiff gait on the left front leg, and swelling in the knee joint (fore knee) (Fig. 1 A) and the dewlap around the neck region (Fig. 1 B). The diagnosis, supported by the smartphone application EDDiE, identified the case as malignant edema. Based on the wound history, physical examination, and clinical signs, the case was tentatively diagnosed as malignant edema and differentially diagnosed as quarter ill, traumatic reticuloperitonitis, and abscess. 2.2. Laboratory examination and findings After a thorough physical and clinical examination, samples were collected from the swollen content in the dewlap using a sterile needle and syringe, and a blood sample was drawn from the jugular vein using an anticoagulant-coated vacutainer. Both samples were placed in a screw-capped vial containing 3 ml of tryptic soy broth and transported immediately to the veterinary laboratory of Debre Markos University, Debre Markos. The samples were enriched overnight and incubated at 37°C in a microbial incubator. After 24 h, both samples were inoculated onto blood agar supplemented with 7% sheep blood under anaerobic conditions. This was achieved by placing a lit candle inside an anaerobic jar to create a low O 2 environment and facilitate CO 2 production. The jar was incubated at 37°C for 48 h. After this period, small gray-white colonies producing a thick hemolytic swarm or dense carpet of organisms were observed (Fig. 2 A). A loopful of pure colonies was used to prepare smears for Gram staining, which revealed large Gram-positive rods (Fig. 2 B), characteristic of Clostridium species. Laboratory tests for oxidase and catalase were negative for this microorganism. Additionally, some of the enriched sample was inoculated onto the same media (blood agar with 7% sheep blood) and incubated at 37°C aerobically for 24 to 48 h, but no growth was observed under these conditions. The images provide crucial visual evidence for diagnosing malignant edema. Figure 2 A shows the typical growth of hemolytic Clostridium colonies under anaerobic conditions, while Fig. 2 B confirms the presence of Gram-positive rods through Gram staining. These laboratory results, combined with the clinical history and physical signs, are essential for confirming the diagnosis of malignant edema in the bovine. 2.3. Management and outcome of the case The treatment and management approach used in this case report involved the administration of intramuscular injections of fortified procaine penicillin at a dose of 28,000 IU/kg body weight, twice daily for three consecutive days. Additionally, dexamethasone sodium phosphate was administered as an anti-inflammatory injection at a dose of 1.5 ml/50 kg body weight, intramuscularly, for three days. The treatment also included the administration of multivitamins and the topical application of iodine following minor surgical drainage of the swelling. The owner was advised to isolate the bull, provide a hygienic bed, and ensure access to clean and fresh water and feed. Unfortunately, despite following this treatment protocol, the bull did not recover. The next morning, upon returning to the clinic for continued therapy, it was found that the animal had died, as reported by the owner. 3. RESULTS AND DISCUSSION The clinical and laboratory findings showed that the case is malignant edema which is a febrile, acute, sporadic, and often fatal disease that affects animals of all ages and species and is transmitted through soil. It is characterized by edema, acute gangrenous inflammation, and toxemia. The disease is caused by one or more pathogenic species of clostridial bacteria, including C. septicum, C. perfringens, C. chauvoei, C. novyi, and C. sordellii, with C. septicum being the most frequently isolated. This bacterium is a normal part of the intestinal flora in animals and is found in feces. Deep wounds and severe tissue trauma create anaerobic conditions that promote the entry and development of the disease. Once in the tissues, histotoxic clostridia produce tissue-degrading enzymes (collagenases, DNases, hyaluronidases, and neuraminidases) and spore-forming and hemolytic toxins, such as α-toxin (from C. septicum), which contribute to the evasion of host immunity, nutrient acquisition, and tissue damage/necrosis. This leads to the proliferation of these microorganisms and the production of toxins, which spread into the systemic circulation, resulting in toxemia, shock, and death (Constable et al., 2017 ; Macheak, 2010 ; Kapustin et al., 2020 ). The characteristics and etiology of the present case are consistent with findings in the literature, and the bull's death can be attributed to toxic shock collapse. Despite the long history of malignant edema in veterinary practice and literature, published information on its occurrence, diagnosis, treatment, control approaches, and economic impact in Ethiopia is limited. In this case report, observed clinical signs included depression, weakness, fever, intoxication, lameness, muscle rigidity, and regional inflammation (heat, edema, pain) at the wound site upon palpation. The local swelling, which initially subsides with direct digital pressure, extended to the surrounding subcutaneous tissues, particularly the dewlap, causing further edema. The bull died within 24 h of the clinical signs being observed at the veterinary center. The characteristic swelling and high mortality rate of this infection justify the term "Malignant Edema". This is consistent with findings and case reports from Gazioglu et al. ( 2018 ), Santos et al. ( 2019 ), Odani et al. ( 2009 )d angelo et al. (2020). The treatment of ME should be considered an emergency due to the acute nature of the disease. Effective treatment typically involves administering high doses of penicillin (crystalline penicillin) intravenously, repeated every 4 to 6 hours, or another broad-spectrum antibiotic along with antitoxin. NSAIDs and supportive therapy, including surgical debridement, are also recommended. Prophylactic antibiotics and vaccination are essential for controlling and preventing the disease (Perdrizet et al., 2012 ; Constable et al., 2017 ; Jahani and Rastabi, 2018 ). This aligns with the treatment protocol in the present case report, where the bull was treated with high doses of fortified procaine penicillin, an antiinflammatory injection of dexamethasone sodium phosphate, multivitamin administration, and topical iodine application. 4. CONCLUSION AND RECOMMENDATIONS This study findings resulted that malignant edema is an acute and often lethal infection characterized by fever, toxemia, and edema, affecting animals of all ages. This study also showed the severity and rapid progression of the disease, which led to the death of the animal within 24 hours. The presence of C. septicum was confirmed through laboratory testing. In Ethiopia, malignant edema causes significant economic losses due to the high mortality rate among livestock. It is crucial to conduct epidemiological investigations to better understand the disease and develop effective management strategies. Implementing control measures, such as antibiotic prophylaxis and vaccination, is recommended, along with training veterinarians and producers in the proper identification and management of the disease. Epidemiological studies are essential for mapping the distribution of the disease and improving diagnostic and treatment methods, thereby protecting animal health in Ethiopia and other affected regions. Declarations The animal owner consented to participate in any diagnostic and therapeutic procedure on his animal. Funding There is no funding organization for this manuscript. It is self-supported. Data Availability declaration No/Not applicable (this manuscript does not report data generation or analysis). Competing Interest Declaration We declared that the authors have no competing interests as defined by BMC, or other interests that might be perceived to influence the results and/or discussion reported in this paper Consent to Publish Declaration Not applicable Ethics Approval and Consent to Participate This work was done for the routine diagnosis of diseases and the affected animal was infected by a disease that was purposefully presented to the veterinary service. Regarding the consent, the owner was volunteer to do all routine examinations on his animal for the confirmation of the case that could help him to control and prevent the spread of the disease. Therefore, ethical approval was not necessary for this type of routine disease diagnosis and treatment procedures. Acknowledgment The authors would like to forward heart felt acknowledgments to the Veterinary clinic and laboratory workers and the owner of the animal who provided adequate information about the outbreak of the case in the surrounding area to control, prevent and eradicate the disease Malignant Edema in the global environment. Authors' Contributions 1. Dessalew Habte: have made substantial contributions to the conception, design of the work, acquisition, analysis, interpretation of data, have drafted the work or substantively revised it. 2. Habtamu Addis: have drafted the work, substantively revised and editing it. References Abdolmohammadi L, Zahmatkesh A (2021) Vaccination against pathogenic clostridia in animals: a review. Trop Anim Health Prod 53(2):1–12 Coetzer J, Tustin R (2014) Infectious Diseases of Livestock, second edition, Cape Town, Oxford University Press, p. 2159 Constable P, Hinchcliff K, Done S, Grundberg W (2017) Veterinary Medicine: A Textbook of the Diseases of Cattle, Horses, Sheep, Pigs, and Goats, eleventh edition, UK: Elsevier. pp. 1648–1660 D’angelo L, Martucciello A, Marianelli C, Grassi C, Schiavo L (2020) Malignant edema case study in postpartum mediterranean buffaloes (Bubalus bubalis). J Buffalo Sci 9(7):1–4 Gazioglu A, Karagülle B, Yüksel H, Keçeci H, Dörtbudak M, Çetinkaya B (2018) Sudden death due to gas gangrene caused by Clostridium septicum in goats. Biomedical Sci Veterinary Res 14(1):1–6 Jahani S, Rastabi H (2018) A Case-Report: Clostridial Edema Following Cesarean section in a Half-Blood Holstein Cattle, A CaseReport. Farm Anim Surg Manage 24(14):1–3 Junior C, Silva R, Lobato F, Navarro M, Uzal F (2020) Gas gangrene in mammals: a review. J Vet Diagn Invest 32(2):175–183 Kapustin A, Laishevtcev A, Ivanov E, Danilyuk V (2020) Species diversity of Clostridia causing malignant edema in cattle. Earth Environ Sci 548(7):1–7 Kusiluka L, Kambarage D, Vetaid K (2016) Diseases of small ruminants: a handbook of common diseases of sheep and goats in sub-Saharan Africa. Centre for Tropical Veterinary Medicine, p 4142 Macheak M (2010) Cattle Clostridial diseases, in Veterinary medicine of small animal clinician. furtherance of Acts of Congress. Miss State Univ 73(2):195–200 Odani J, Blanchard P, Adaska J, Moeller R, Uzal F (2009) Malignant edema in postpartum dairy cattle. J Vet Diagn Invest 21(6):920–924 Parish M, Hodgson D, Valberg S (1996) Clostridial myonecrosis, in large animal internal medicine, second edition, Mosby-Year Book of St. Louis. Pp. 1504–1509 Perdrizet J, Callihan D, Rebhun W, Shin S (2012) Successful management of malignant edema caused by Clostridium septicum in a horse. Cornell Veterinary 77(4):328–338 Raymundo L, Bandarra M, Boabaid M, Raymundo D, Bandarra P, Boabaid F (2014) Clostridial Diseases Diagnosed in Herbivores in Southern Brazil. Acta Sci Veterinariae 42:2–9 Roger W, Blowey A, David W (2011) Malignant edema, in Color Atlas of Diseases and Disorders of Cattle, third edition, Wiley Blackwell, pp.272–286 Santos B, Ladeira S, Riet F, Soares M, Marcolongo C (2019) Clostridial diseases diagnosed in cattle from the South of Rio Grande do Sul, Brazil. A forty-year survey (1978–2018) and a brief review of the literature1. Pesquisa Vet Brasileira 39(7):435446 Silva R, Uzal A, Oliveira C, Lobato F (2016) Gas Gangrene (Malignant Edema). Clostridial diseases of animals. Wiley Blackwell, p 24354 Additional Declarations The authors declare no competing interests. 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INTRODUCTION","content":"\u003cp\u003eMalignant edema (ME; gas gangrene) is an acute, often fatal wound infection caused by one or more pathogenic species of Clostridium. It affects animals of all ages and grazing species, and is globally distributed, particularly in areas with soil heavily contaminated by bacterial spores. It is considered an \"exogenous\" infection because environmental bacterial microorganisms penetrate tissues through wounds in the skin or mucous membranes, creating an anaerobic environment (Silva et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Kusiluka et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe disease malignant edema is primarily caused by \u003cem\u003eClostridium septicum\u003c/em\u003e, an anaerobic, Gram-positive, spore-forming organism. Although \u003cem\u003eC. septicum\u003c/em\u003e is the main cause of edematous and emphysematous conditions, other Clostridium species, such as Clostridium chauvoei, Clostridium perfringens, Clostridium sordellii, and Clostridium novyi, can also be associated with the disease, indicating mixed infections. C. septicum and other clostridia, along with their spores, are found in organic-rich, high-humidity soils and in the intestinal tracts of animals, making them ubiquitous worldwide. Infection usually occurs through the contamination of wounds caused by accidents such as castration, sterilization, shearing, unhygienic vaccination, tail docking, and parturition, which activate dormant spores in the soil (Parish et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1996\u003c/span\u003e; Coetzer and Tustin, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Kapustin et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe disease is typically acute, though subacute and chronic cases may occur sporadically. Clinical signs of malignant edema include anorexia, depression, weakness, and local lesions at the infection site characterized by soft, hot, and pasty swelling with marked local erythema and intense pain upon palpation. In advanced stages, the swelling becomes tense and dark, with gaseous and edematous accumulations that can be palpated as crepitus in the subcutaneous tissue around the infected area. Other signs include high fever (41 to 42\u0026deg;C), intoxication, muscle tremors, lameness, rigidity, and recumbency. The skin may appear tense and diffusely black or red with bruising and suffusions. In advanced stages, subnormal body temperature is common, and the affected area may become cold with the disappearance of pain due to necrosis of local nerve endings. Death often results from systemic shock and toxemia within a few hours to 2\u0026ndash;4 days after the onset of clinical signs (Constable et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; D'Angelo et al., 2020; Junior et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe diagnosis of malignant edema is based on characteristic clinical signs, a history of wounds, and laboratory examination using various techniques. Diagnostic confirmation can be achieved through fluorescent antibody staining of tissue smears and by observing clinicopathological changes at necropsy, such as dark red, swollen muscles filled with hemorrhagic proteinaceous exudate and minimal gas production. ME can be differentially diagnosed from other conditions, including abscesses, quarter ill, snake bites, and anthrax (Santos et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Raymundo et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eEffective management of malignant edema involves proper wound management and the use of antibiotics such as penicillin and tetracycline. Supportive treatments, including fluid therapy, anti-inflammatory agents, and proper nutrition, are also important. Maintaining good husbandry and sanitary practices during invasive procedures; such as shearing, castration, surgical and obstetric manipulations, tail docking, and injection administration is essential for preventing malignant edema. In endemic areas, immunization with multivalent vaccines combining \u003cem\u003eC. septicum and C. chauvoei\u003c/em\u003e, especially before procedures like dehorning, castration, or tail docking, is crucial (Abdolmohammadi and Zahmatkesh, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Coetzer and Tustin, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eMalignant edema causes significant economic losses due to reduced productivity and reproductive capacity, as well as the costs of treatment, control, and prevention, alongside animal morbidity and mortality in Ethiopia. All livestock species, regardless of age, are at risk of infection from endemic and/or sporadic strains of this bacterial disease, primarily caused by \u003cem\u003eC. septicum\u003c/em\u003e in cattle, with no clear framework for diagnosis, treatment, and preventive measures with available vaccines (Roger et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Gazioglu et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This case report describes an episode of malignant edema caused by Clostridium bacteria in an adult local breed bovine.\u003c/p\u003e"},{"header":"2. MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Case Description\u003c/h2\u003e \u003cp\u003eAn adult local breed bull, weighing 290 kg, was presented to the Veterinary Clinic of Debre Markos in Debre Markos town, Ethiopia, on April 4, 2023, with severe symptoms including depression, anorexia, lameness, weakness, and swelling in the left forequarter, followed by subcutaneous edema in the neck region. The swelling developed after a wound of unknown cause, which occurred approximately one week prior to the clinic visit.\u003c/p\u003e \u003cp\u003eThe physical examination revealed a respiratory rate of 32 breaths/minute, a body temperature of 41.7\u0026deg;C, a heart rate of 84 beats/min, and a soft, hot, and pasty swelling that indented upon palpation. Initially, the swelling was localized in the knee joint and later extended to the subcutaneous tissue of the dewlap, with marked local erythema and intense pain at the site of infection. The skin was stretched, diffusely red or black, with bruising and suffusions. The clinical examination also noted weakness, anorexia, depression, salivation, intoxication, high fever (41.7\u0026deg;C), lameness with a stiff gait on the left front leg, and swelling in the knee joint (fore knee) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA) and the dewlap around the neck region (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe diagnosis, supported by the smartphone application EDDiE, identified the case as malignant edema. Based on the wound history, physical examination, and clinical signs, the case was tentatively diagnosed as malignant edema and differentially diagnosed as quarter ill, traumatic reticuloperitonitis, and abscess.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Laboratory examination and findings\u003c/h2\u003e \u003cp\u003eAfter a thorough physical and clinical examination, samples were collected from the swollen content in the dewlap using a sterile needle and syringe, and a blood sample was drawn from the jugular vein using an anticoagulant-coated vacutainer. Both samples were placed in a screw-capped vial containing 3 ml of tryptic soy broth and transported immediately to the veterinary laboratory of Debre Markos University, Debre Markos. The samples were enriched overnight and incubated at 37\u0026deg;C in a microbial incubator.\u003c/p\u003e \u003cp\u003eAfter 24 h, both samples were inoculated onto blood agar supplemented with 7% sheep blood under anaerobic conditions. This was achieved by placing a lit candle inside an anaerobic jar to create a low O \u003csub\u003e2\u003c/sub\u003e environment and facilitate CO\u003csub\u003e2\u003c/sub\u003e production. The jar was incubated at 37\u0026deg;C for 48 h. After this period, small gray-white colonies producing a thick hemolytic swarm or dense carpet of organisms were observed (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). A loopful of pure colonies was used to prepare smears for Gram staining, which revealed large Gram-positive rods (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB), characteristic of Clostridium species. Laboratory tests for oxidase and catalase were negative for this microorganism.\u003c/p\u003e \u003cp\u003eAdditionally, some of the enriched sample was inoculated onto the same media (blood agar with 7% sheep blood) and incubated at 37\u0026deg;C aerobically for 24 to 48 h, but no growth was observed under these conditions. The images provide crucial visual evidence for diagnosing malignant edema. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA shows the typical growth of hemolytic Clostridium colonies under anaerobic conditions, while Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB confirms the presence of Gram-positive rods through Gram staining. These laboratory results, combined with the clinical history and physical signs, are essential for confirming the diagnosis of malignant edema in the bovine.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Management and outcome of the case\u003c/h2\u003e \u003cp\u003eThe treatment and management approach used in this case report involved the administration of intramuscular injections of fortified procaine penicillin at a dose of 28,000 IU/kg body weight, twice daily for three consecutive days. Additionally, dexamethasone sodium phosphate was administered as an anti-inflammatory injection at a dose of 1.5 ml/50 kg body weight, intramuscularly, for three days. The treatment also included the administration of multivitamins and the topical application of iodine following minor surgical drainage of the swelling. The owner was advised to isolate the bull, provide a hygienic bed, and ensure access to clean and fresh water and feed. Unfortunately, despite following this treatment protocol, the bull did not recover. The next morning, upon returning to the clinic for continued therapy, it was found that the animal had died, as reported by the owner.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. RESULTS AND DISCUSSION","content":"\u003cp\u003eThe clinical and laboratory findings showed that the case is malignant edema which is a febrile, acute, sporadic, and often fatal disease that affects animals of all ages and species and is transmitted through soil. It is characterized by edema, acute gangrenous inflammation, and toxemia. The disease is caused by one or more pathogenic species of clostridial bacteria, including \u003cem\u003eC. septicum, C. perfringens, C. chauvoei, C. novyi, and C. sordellii, with C. septicum\u003c/em\u003e being the most frequently isolated. This bacterium is a normal part of the intestinal flora in animals and is found in feces. Deep wounds and severe tissue trauma create anaerobic conditions that promote the entry and development of the disease. Once in the tissues, histotoxic clostridia produce tissue-degrading enzymes (collagenases, DNases, hyaluronidases, and neuraminidases) and spore-forming and hemolytic toxins, such as α-toxin (from C. septicum), which contribute to the evasion of host immunity, nutrient acquisition, and tissue damage/necrosis. This leads to the proliferation of these microorganisms and the production of toxins, which spread into the systemic circulation, resulting in toxemia, shock, and death (Constable et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Macheak, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Kapustin et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The characteristics and etiology of the present case are consistent with findings in the literature, and the bull's death can be attributed to toxic shock collapse.\u003c/p\u003e \u003cp\u003eDespite the long history of malignant edema in veterinary practice and literature, published information on its occurrence, diagnosis, treatment, control approaches, and economic impact in Ethiopia is limited. In this case report, observed clinical signs included depression, weakness, fever, intoxication, lameness, muscle rigidity, and regional inflammation (heat, edema, pain) at the wound site upon palpation. The local swelling, which initially subsides with direct digital pressure, extended to the surrounding subcutaneous tissues, particularly the dewlap, causing further edema. The bull died within 24 h of the clinical signs being observed at the veterinary center. The characteristic swelling and high mortality rate of this infection justify the term \"Malignant Edema\". This is consistent with findings and case reports from Gazioglu et al. (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), Santos et al. (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), Odani et al. (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2009\u003c/span\u003e)d angelo et al. (2020).\u003c/p\u003e \u003cp\u003eThe treatment of ME should be considered an emergency due to the acute nature of the disease. Effective treatment typically involves administering high doses of penicillin (crystalline penicillin) intravenously, repeated every 4 to 6 hours, or another broad-spectrum antibiotic along with antitoxin. NSAIDs and supportive therapy, including surgical debridement, are also recommended. Prophylactic antibiotics and vaccination are essential for controlling and preventing the disease (Perdrizet et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Constable et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Jahani and Rastabi, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This aligns with the treatment protocol in the present case report, where the bull was treated with high doses of fortified procaine penicillin, an antiinflammatory injection of dexamethasone sodium phosphate, multivitamin administration, and topical iodine application.\u003c/p\u003e"},{"header":"4. CONCLUSION AND RECOMMENDATIONS","content":"\u003cp\u003eThis study findings resulted that malignant edema is an acute and often lethal infection characterized by fever, toxemia, and edema, affecting animals of all ages. This study also showed the severity and rapid progression of the disease, which led to the death of the animal within 24 hours. The presence of \u003cem\u003eC. septicum\u003c/em\u003e was confirmed through laboratory testing. In Ethiopia, malignant edema causes significant economic losses due to the high mortality rate among livestock. It is crucial to conduct epidemiological investigations to better understand the disease and develop effective management strategies. Implementing control measures, such as antibiotic prophylaxis and vaccination, is recommended, along with training veterinarians and producers in the proper identification and management of the disease. Epidemiological studies are essential for mapping the distribution of the disease and improving diagnostic and treatment methods, thereby protecting animal health in Ethiopia and other affected regions.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eThe animal owner consented to participate in any diagnostic and therapeutic procedure on his animal.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere is no funding organization for this manuscript. It is self-supported.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo/Not applicable (this manuscript does not report data generation or analysis).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interest Declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe declared that the authors have no competing interests as defined by BMC, or other interests that might be perceived to influence the results and/or discussion reported in this paper\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Publish Declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics Approval and Consent to Participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was done for the routine diagnosis of diseases and the affected animal was infected by a disease that was purposefully presented to the veterinary service. Regarding the consent, the owner was volunteer to do all routine examinations on his animal for the confirmation of the case that could help him to control and prevent the spread of the disease. Therefore, ethical approval was not necessary for this type of routine disease diagnosis and treatment procedures.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to forward heart felt acknowledgments to the Veterinary clinic and laboratory workers and the owner of the animal who provided adequate information about the outbreak of the case in the surrounding area to control, prevent and eradicate the disease Malignant Edema in the global environment.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors' Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e1. Dessalew Habte: have made substantial contributions to the conception, design of the work, acquisition, analysis, interpretation of data, have drafted the work or substantively revised it.\u003c/p\u003e\n\u003cp\u003e2. Habtamu Addis: have drafted the work, substantively revised and editing it.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbdolmohammadi L, Zahmatkesh A (2021) Vaccination against pathogenic clostridia in animals: a review. Trop Anim Health Prod 53(2):1\u0026ndash;12\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCoetzer J, Tustin R (2014) Infectious Diseases of Livestock, second edition, Cape Town, Oxford University Press, p. 2159\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eConstable P, Hinchcliff K, Done S, Grundberg W (2017) Veterinary Medicine: A Textbook of the Diseases of Cattle, Horses, Sheep, Pigs, and Goats, eleventh edition, UK: Elsevier. pp. 1648\u0026ndash;1660\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eD\u0026rsquo;angelo L, Martucciello A, Marianelli C, Grassi C, Schiavo L (2020) Malignant edema case study in postpartum mediterranean buffaloes (Bubalus bubalis). J Buffalo Sci 9(7):1\u0026ndash;4\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGazioglu A, Karag\u0026uuml;lle B, Y\u0026uuml;ksel H, Ke\u0026ccedil;eci H, D\u0026ouml;rtbudak M, \u0026Ccedil;etinkaya B (2018) Sudden death due to gas gangrene caused by Clostridium septicum in goats. Biomedical Sci Veterinary Res 14(1):1\u0026ndash;6\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJahani S, Rastabi H (2018) A Case-Report: Clostridial Edema Following Cesarean section in a Half-Blood Holstein Cattle, A CaseReport. Farm Anim Surg Manage 24(14):1\u0026ndash;3\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJunior C, Silva R, Lobato F, Navarro M, Uzal F (2020) Gas gangrene in mammals: a review. J Vet Diagn Invest 32(2):175\u0026ndash;183\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKapustin A, Laishevtcev A, Ivanov E, Danilyuk V (2020) Species diversity of Clostridia causing malignant edema in cattle. Earth Environ Sci 548(7):1\u0026ndash;7\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKusiluka L, Kambarage D, Vetaid K (2016) Diseases of small ruminants: a handbook of common diseases of sheep and goats in sub-Saharan Africa. Centre for Tropical Veterinary Medicine, p 4142\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMacheak M (2010) Cattle Clostridial diseases, in Veterinary medicine of small animal clinician. furtherance of Acts of Congress. Miss State Univ 73(2):195\u0026ndash;200\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOdani J, Blanchard P, Adaska J, Moeller R, Uzal F (2009) Malignant edema in postpartum dairy cattle. J Vet Diagn Invest 21(6):920\u0026ndash;924\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eParish M, Hodgson D, Valberg S (1996) Clostridial myonecrosis, in large animal internal medicine, second edition, Mosby-Year Book of St. Louis. Pp. 1504\u0026ndash;1509\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePerdrizet J, Callihan D, Rebhun W, Shin S (2012) Successful management of malignant edema caused by Clostridium septicum in a horse. Cornell Veterinary 77(4):328\u0026ndash;338\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRaymundo L, Bandarra M, Boabaid M, Raymundo D, Bandarra P, Boabaid F (2014) Clostridial Diseases Diagnosed in Herbivores in Southern Brazil. Acta Sci Veterinariae 42:2\u0026ndash;9\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRoger W, Blowey A, David W (2011) Malignant edema, in Color Atlas of Diseases and Disorders of Cattle, third edition, Wiley Blackwell, pp.272\u0026ndash;286\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSantos B, Ladeira S, Riet F, Soares M, Marcolongo C (2019) Clostridial diseases diagnosed in cattle from the South of Rio Grande do Sul, Brazil. A forty-year survey (1978\u0026ndash;2018) and a brief review of the literature1. Pesquisa Vet Brasileira 39(7):435446\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSilva R, Uzal A, Oliveira C, Lobato F (2016) Gas Gangrene (Malignant Edema). Clostridial diseases of animals. Wiley Blackwell, p 24354\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Bahir Dar University","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Anaerobic infection, cattle, gas gangrene, toxemia, veterinary diagnosis","lastPublishedDoi":"10.21203/rs.3.rs-6391174/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6391174/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study documents a case of malignant edema in an adult local breed bovine in Ethiopia. The objective was to detail the clinical presentation, diagnosis, and treatment of malignant edema caused by \u003cem\u003eClostridium septicum.\u003c/em\u003e The affected animal, weighing 290 kg, exhibited symptoms including depression, anorexia, lameness, weakness, and swelling in the left forequarter, as well as edema in the dewlap region. The methodology involved a comprehensive clinical examination, bacterial culture, and Gram staining to confirm the presence of \u003cem\u003eC. septicum.\u003c/em\u003e Clinical findings included an elevated respiratory rate, fever (41.7\u0026deg;C), and a hot, soft, and pasty edema initially localized in the knee joint, which subsequently extended to the dewlap. The affected area showed erythema and was intensely painful. Despite treatment with dexamethasone sodium phosphate, high doses of fortified procaine penicillin, multivitamin injections, and topical iodine application, the intervention proved ineffective, leading to the animal's death. This case underscores the need for further clinical and epidemiological research to enhance management and prevention strategies for malignant edema in the region.\u003c/p\u003e","manuscriptTitle":"Clinical and Laboratory Analysis of Malignant Edema (Gas Gangrene) in Cattle in Ethiopia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-10 08:02:08","doi":"10.21203/rs.3.rs-6391174/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a01512d8-ec77-4f94-a36d-873504c03df8","owner":[],"postedDate":"April 10th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":46941149,"name":"Large Animal Medicine"}],"tags":[],"updatedAt":"2025-04-10T08:02:08+00:00","versionOfRecord":[],"versionCreatedAt":"2025-04-10 08:02:08","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6391174","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6391174","identity":"rs-6391174","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}