RESUMO

Tetanus is an infectious disease caused by a ubiquitous bacterium Clostridium tetani, that synthesizes and releasesa potent neurotoxin under anaerobic conditions, which is responsible for the clinical manifestations. As it is found in soil contaminated with animal and human excreta, it is difficult to eradicate but it may be prevented by immunization. Immunization rate has decreased in the last years, especially during the COVID-19 pandemic. We report two cases of tetanus, attended during 2022. A 39-year-old man whose entry route was a gunshot wound and he was discharged from the intensive care unit (ICU) and a second case of an 83-year-old woman with unknown entry point, who died during her ICU stay. The cases reported highlight that it is a life-threatening disease, its diagnosis is mainly clinical and it should be in the algorithm of differential diagnoses. We emphasize about the prompt treatment administration or consultation to a specialized healthcare center. The importance of this presentation is to show the severity of the disease, whose assessment is mainly clinical and should not escape the algorithm of differential diagnoses, emphasizing that treatment should be instituted early or when in doubt consult a specialized center. In addition to this, it is important to check theimmunization rate in our country, especially during thepandemic, becauseit is a vaccine-preventable disease.

El tétanos es causado por el Clostridium tetani, una bacteria ubicua que frente a condiciones de anaerobiosis puede sintetizar y liberar sus toxinas, responsables del cuadro clínico. Dado que es una bacteria que se encuentra en el suelo y en el tracto gastrointestinal de muchas especies, se trata de una enfermedad no erradicable pero si controlable a través de la inmunización para la prevención. Las tasas de inmunización han disminuido en los últimos años, evento que se ha acentuado durante la COVID-19. Se presentan a continuación dos casos clínicos ingresados durante el año 2022. El primero es un hombre de 39 años cuya puerta de entrada fue una herida de arma de fuego conevolución favorable y el segundo caso se trata de una mujer de 83 años sin puerta de entrada clara quien falleció durante su internación en terapia intensiva. La importancia de esta presentación es mostrar la gravedad de la enfermedad, cuyavaloración es principalmente clínica y no debe escapar al algoritmo de diagnósticos diferenciales, acentuando que se debe instaurar el tratamiento de forma precoz o frente a la duda consultar con un centro especializado. Asímismo, es importante revisar las tasas de inmunización en nuestro país y los cambios que se presentaron durante la pandemia, teniendo en cuenta, como se ha expuesto previamente, se trata de una enfermedad inmunoprevenible.

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Tetanus is a serious and rare disease in small animals, of an acute nature and mediated by the action of the neurotoxin tetanospasmin, from the bacillus Clostridium tetani. This report describes a case of tetanus in a canine, with emphasis on its clinical and therapeutic aspects, in addition to its clinical recovery. A canine, female, Pitbull breed, five months old, in status epilepticus, was attended at the Veterinary Hospital of the University of Passo Fundo. After stabilizing the patient, the animal was found in a trestle position, with generalized spastic paralysis, risus sardonicus, trismus, erect tail and ears, and difficulty in expanding the thorax. The presumptive diagnosis of tetanus was established based on the anamnesis, clinical signs, and laboratory findings. The established supportive treatment included hydro electrolytic replacement, antitetanus serum, antibiotic therapy, analgesia, myorelaxant and anticonvulsant drugs. The patient's intensive management was carried out with urethral and gastroesophageal probing, changes of position every two hours, and reduction of environmental stimuli. Physiotherapy and acupuncture were also used to complement the treatment and accelerate recovery. The patient was discharged one month after the beginning of the treatment, presenting satisfactory evolution. In the present case, the clinical examination in association with the characteristic clinical signs of the disease, added to the detailed anamnesis, was essential for the presumptive diagnosis of tetanus. In addition, intensive management, drug treatment, physiotherapy, and acupuncture enabled the evolution of the clinical condition to cure.

O tétano é uma doença grave e rara em pequenos animais, de caráter agudo e mediada pela ação da neurotoxina tetanospasmina, proveniente do bacilo Clostridium tetani. Neste relato descreve-se um caso de tétano em um canino, com ênfase em seus aspectos clínicos e terapêuticos, além da sua recuperação clínica. Foi atendido no Hospital Veterinário da Universidade de Passo Fundo, um canino, fêmea, da raça Pitbull, com cinco meses, em status epilepticus. Após a estabilização da paciente, constataram-se animal em posição de cavalete, com paralisia espástica generalizada, risus sardonicus, trismo, cauda e orelhas eretas e dificuldade em expandir o tórax. O diagnóstico presuntivo de tétano foi firmado a partir da anamnese, sinais clínicos e achados laboratoriais. O tratamento suporte estabelecido incluiu reposição hidroeletrolítica, soro antitetânico, antibioticoterapia, analgesia, fármacos miorrelaxantes e anticonvulsivantes. O manejo intensivo da paciente foi realizado com sondagem uretral e nasoesofágica, trocas de decúbito a cada duas horas e diminuição de estímulos ambientais. A fisioterapia e a acupuntura também foram utilizadas para complementar o tratamento e acelerar a recuperação. A paciente recebeu alta médica um mês após o início do tratamento, apresentando evolução satisfatória. No presente caso, o exame clínico em associação aos sinais clínicos característicos da doença, acrescidos da anamnese minuciosa foram fundamentais ao diagnóstico presuntivo de tétano. Além disso, o manejo intensivo, o tratamento medicamentoso, assim como a realização de fisioterapia e acupuntura, possibilitaram a evolução do quadro clínico à cura.

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RESUMEN INTRODUCCIÓN: El tétanos es una enfermedad inmunoprevenible, ocasionada por la bacteria Clostridium tetani, desencadenando una enfermedad caracterizada por espasmos musculares, insuficiencia respiratoria y disautonomías, potencialmente mortal. MATERIALES Y MÉTODOS: Presentamos una serie de tres pacientes que consultaron al servicio de urgencias por presentar trismus, rigidez muscular generalizada y dificultad respiratoria, requiriendo manejo en la unidad de cuidados intensivos, con relajación muscular y administración intramuscular e intratecal de inmunoglobulina antitetánica, con evolución satisfactoria en todos los casos. DISCUSIÓN: Su tratamiento está divido en dos grandes secciones; la primera parte, el control de la infección y eliminación del agente causal, con lavado y desbridamiento de heridas, administración de antibióticos y neutralización de la neurotoxina. La segunda parte del tratamiento está en el soporte vital en la unidad de cuidados intensivos, con la administración de sedación, relajación muscular, control de disautonomías y manejo de complicaciones. CONCLUSIONES: El tétanos a pesar de los avances en vacunación aún es una enfermedad presente, cuyo diagnóstico y tratamiento rápido y adecuado, permite sobrevivir a los pacientes, como en los casos aquí reportados.

ABSTRACT INTRODUCTION: Tetanus is an immuno-preventable disease, produced by the bacterium Clostridium tetani, that causes a disease characterized by muscle spasms, respiratory insufficiency and life-threatening dysautonomia. MATERIALS AND METHODS: We present a series of three patients who consulted for trismus, muscle stiffness and respiratory failure, which required intensive care management, muscle relaxation, intramuscular and intrathecal administration of tetanus immu-noglobulin, with satisfactory outcomes in all the cases. DISCUSSION: Its treatment is divided into two main sections; the first part, the control of infection and elimination of the causative agent, with washing and debridement of wounds, administration of antibiotics and neutralization of the neurotoxin. The second part is life support in the intensive care unit, with the administration of sedation, muscular relaxation and control of dysautonomia and the management of complications. CONCLUSIONS: Despite the advances in vaccination, tetanus is still a present disease, whose diagnosis and rapid and adequate treatment allows patients to survive, as in the cases reported here.

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RESUMEN Fundamento: El tétanos es una enfermedad infecciosa causada por las esporas de la bacteria Clostridium tetani que afecta el sistema nervioso. La inmunización contra esta enfermedad ha permitido reducir la morbilidad y la mortalidad en Cuba por esa causa. Los pocos casos reportados son pacientes sin previa vacunación contra el tétanos, por lo que constituye de interés para la comunidad científica el reporte de aquellos que desarrollan esa enfermedad después de haber sido vacunados. Objetivo: Describir el caso de un paciente con diagnóstico de tétanos generalizado a pesar de que ya había sido vacunado contra esa enfermedad. Presentación del caso: Paciente de 33 años, con el antecedente de haber sido vacunado contra el tétanos, al cual le diagnosticaron tétanos generalizado, por lo se decidió su ingresó en la unidad de cuidados intensivos. Clínicamente se manifestó por contracciones musculares generalizadas y rigidez, los síntomas y signos neurológicos tuvieron una duración de 4 semanas. A pesar de las complicaciones el paciente egresó con adecuada calidad de vida. Conclusiones: La presentación de este caso busca concientizar en la comunidad científica la importancia de considerar el diagnóstico de tétanos en pacientes ya vacunados, lo cual es poco frecuente y obliga a actuar con prontitud.

ABSTRACT Background: Tetanus is an infectious disease caused by the bacterium Clostridium tetani spores which affects the nervous system. In Cuba, immunization against this disease has made it possible to reduce morbidity and mortality from this cause. The few cases reported are patients without prior vaccination against tetanus, so is the interest to the scientific community the reporting of those who develop this disease after having been vaccinated. Objective: To describe a patient´s case with a generalized tetanus diagnosis despite has already been vaccinated against this disease. Case report: A 33-year-old patient, with a history of has been vaccinated against tetanus, and diagnosed with generalized tetanus, so it was decided he to be admitted to the intensive care unit. Clinically manifested by generalized muscle contractions and stiffness, the neurological symptoms and signs lasted 4 weeks. Despite the complications, the patient discharged from hospital with an adequate life quality. Conclusions: This case presentation searches for raising awareness in the scientific community about the importance of considering the diagnosis of tetanus in previous vaccinated patients, which is rare and requires quickness of action.

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O tétano é uma doença infecciosa não contagiosa, desencadeada pela ação de neurotoxinas produzidas pela bactéria Clostridium tetani. Dentre as espécies mais suscetíveis e de maior ocorrência em estudos epidemiológicos, destacam-se os equinos. Neste estudo de caso, foi atendido no Hospital Veterinário do IFPB campus Sousa, uma fêmea equina, SRD, 8 anos de idade, no 6º mês de gestação. O animal apresentava taquipneia, taquicardia, espasticidade dos membros, protrusão da terceira pálpebra, hiperestesia, cauda em bandeira, rigidez da musculatura do abdome e discreta rigidez da musculatura cervical. Com a intervenção medicamentosa baseada no uso de soro antitetânico, antibioticoterapia com benzilpenicilina benzatina, acepromazina e fluidoterapia à base de solução de ringer com lactato, associado ao repouso em ambiente silencioso e termicamente agradável, obteve-se resultados satisfatórios nos primeiros dias do inicio do tratamento e recuperação total após o 15º dia. Além disso, ao 15°dia pós-internamento e antecedendo a alta do animal foi realizada a avaliação ultrassonográfica transretal, confirmando a viabilidade fetal.(AU)

Tetanus is a non-contagious infectious disease triggered by neurotoxins produced by the bacterium Clostridium tetani. Horses are among the most susceptible species and also are frequent targets in epidemiological studies. In this case study, a mare, crossbred, 8 years old, in the sixth month of gestation, was assisted at the Veterinary Hospital at IFPB Sousa campus. The animal was presenting some clinical signs, such as tachypnea, tachycardia, limb spasticity, protrusion of the third eyelid, hyperesthesia, elevated tail, stiffness of the abdomen muscles and slight stiffness of the cervical muscles. Medicines treatment was based on the use of tetanus antitoxin, benzathine penicillin antibiotic therapy, acepromazine and lactate ringer's solution therapy, combined with a calm and thermally pleasant environment. Satisfactory results were obtained from the first days of treatment and full recovery of the animal after the 15th day of hospitalization. In addition, on the 15thday after admission at the hospital and before discharge, a transrectal ultrasound evaluation was performed, confirming fetal viability.(AU)

El tétano es uma enfermedad infecciosa desencadenada por la acción de las neurotoxinas producidas por la bacteria Clostridium tetani. Entre las especies más susceptibles y más frecuentes en estudios epidemiológicos, destacan los caballos. En este estudio de caso, fue vista en el Hospital Veterinario Campus Sousa de IFPB, una yegua preñada, SRD, de 8 años de edad, en el sexto mes degestación. El animal presentaba taquipnea, taquicardia, espasticidad, posición del caballete, protrusión del tercer párpado, hiperestesia, colade bandera, rigidez de los músculos del abdomen y leve rigidez de los músculos cervicales. Con la intervención farmacológica basada en el uso del suero antitetánico, la terapia con antibióticos benzatínicos y bencilpenicilinas, la acepromacina y la fluidoterapia basada en la solución de Ringer lactato, asociada con un descanso tranquilo y placentero térmicamente, se obtuvieron resultados satisfactorios en los primeros días de inicio del tratamiento. y recuperación completa después del día 15. Además, el día 15 después del ingreso y antes del alta del animal, se realizó una evaluación ecográfica transrectal, confirmando la viabilidad fetal.(AU)

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RESUMO

O tétano é uma doença infecciosa não contagiosa, desencadeada pela ação de neurotoxinas produzidas pela bactéria Clostridium tetani. Dentre as espécies mais suscetíveis e de maior ocorrência em estudos epidemiológicos, destacam-se os equinos. Neste estudo de caso, foi atendido no Hospital Veterinário do IFPB campus Sousa, uma fêmea equina, SRD, 8 anos de idade, no 6º mês de gestação. O animal apresentava taquipneia, taquicardia, espasticidade dos membros, protrusão da terceira pálpebra, hiperestesia, cauda em bandeira, rigidez da musculatura do abdome e discreta rigidez da musculatura cervical. Com a intervenção medicamentosa baseada no uso de soro antitetânico, antibioticoterapia com benzilpenicilina benzatina, acepromazina e fluidoterapia à base de solução de ringer com lactato, associado ao repouso em ambiente silencioso e termicamente agradável, obteve-se resultados satisfatórios nos primeiros dias do inicio do tratamento e recuperação total após o 15º dia. Além disso, ao 15°dia pós-internamento e antecedendo a alta do animal foi realizada a avaliação ultrassonográfica transretal, confirmando a viabilidade fetal.

Tetanus is a non-contagious infectious disease triggered by neurotoxins produced by the bacterium Clostridium tetani. Horses are among the most susceptible species and also are frequent targets in epidemiological studies. In this case study, a mare, crossbred, 8 years old, in the sixth month of gestation, was assisted at the Veterinary Hospital at IFPB Sousa campus. The animal was presenting some clinical signs, such as tachypnea, tachycardia, limb spasticity, protrusion of the third eyelid, hyperesthesia, elevated tail, stiffness of the abdomen muscles and slight stiffness of the cervical muscles. Medicines treatment was based on the use of tetanus antitoxin, benzathine penicillin antibiotic therapy, acepromazine and lactate ringer's solution therapy, combined with a calm and thermally pleasant environment. Satisfactory results were obtained from the first days of treatment and full recovery of the animal after the 15th day of hospitalization. In addition, on the 15thday after admission at the hospital and before discharge, a transrectal ultrasound evaluation was performed, confirming fetal viability.

El tétano es uma enfermedad infecciosa desencadenada por la acción de las neurotoxinas producidas por la bacteria Clostridium tetani. Entre las especies más susceptibles y más frecuentes en estudios epidemiológicos, destacan los caballos. En este estudio de caso, fue vista en el Hospital Veterinario Campus Sousa de IFPB, una yegua preñada, SRD, de 8 años de edad, en el sexto mes degestación. El animal presentaba taquipnea, taquicardia, espasticidad, posición del caballete, protrusión del tercer párpado, hiperestesia, colade bandera, rigidez de los músculos del abdomen y leve rigidez de los músculos cervicales. Con la intervención farmacológica basada en el uso del suero antitetánico, la terapia con antibióticos benzatínicos y bencilpenicilinas, la acepromacina y la fluidoterapia basada en la solución de Ringer lactato, asociada con un descanso tranquilo y placentero térmicamente, se obtuvieron resultados satisfactorios en los primeros días de inicio del tratamiento. y recuperación completa después del día 15. Además, el día 15 después del ingreso y antes del alta del animal, se realizó una evaluación ecográfica transrectal, confirmando la viabilidad fetal.

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Tétano é uma doença infecciosa causada por toxinas do Clostridium tetani, um bacilo Gram-positivo, esporulado, anaeróbico obrigatório, que habita normalmente o trato gastrointestinal de herbívoros e também o solo. A bactéria produz as toxinas tetanosespasmina (neurotoxina), hemolisina (tetanolisina) e toxina não-espasmogênica de ação periférica que são responsáveis pelo quadro clínico observado nos animais. Essa também é encontrada no intestino dos equinos, sendo considerada uma bactéria natural desses animais. Foi atendido um muar, macho de 12 anos de idade, peso de 250 kg, no exame clínico o animal apresentava andar em forma de cavalete, calda em bandeira, quando movimentava a cabeça apresentava protrusão da terceira pálpebra, foi examinado os cascos, bem como todo o corpo do animal e não foi encontrado nenhum ferimento, com base no exame físico do animal, bem como os sinais clínicos apresentados, o diagnóstico de tétano foi estabelecido. O tratamento preconizado foi imediato instituído com soro antitetânico por via intravenosa, terapia com antibióticos e a terapia com tranquilizantes. Após 10 dias do início do tratamento, o animal apresentava diminuição da sintomatologia da enfermidade, retornando todos os parâmetros à normalidade, e ficando em observação por mais 10 dias, apresentando cura clínica.

Tetanus is an infectious disease caused by Clostridium tetani toxins, a bacterum anaerobic Gram positive, sporulated bacillus that normally inhabits the gastrointestinal tract of herbivores and also the soil. The bacterium produces the toxins tetanosespasmin (neurotoxin), hemolysin (tetanolysin) and non-spasmo genictoxin of peripheral action that are responsible for the clinical picture observed in the animals. This is also found in the intestines of horses, being considered a natural bacterium of these animals. A mule, a 12-year-old male, weighing 250 kg, was attended at the clinical examination. The animal presented a pallet-shapedwalk, flag syringe, while the head was protruding from the third eyelid, the hoofs were examined, as well as the entire body of the animal and no injury was found, based on the physical examination of the animal as well as the clinical signs presented the diagnosis of tetanus was established. The recommended treatment was immediately instituted with intravenous anti-tetanus serum, antibiotictherapy and tranquilizer therapy. Ten days after the start of treatment, the animal presented decrease of the symptomatology of the disease, returning all parameters to normality, and remaining in observation for another 10 days, presenting clinical cure.

Tétano es una enfermedad infecciosa causada por toxinas del Clostridium tetani, un bacilo Gram-positivo, esporulado, anaeróbico obligatorio, que habita normalmente el tracto gastrointestinal de herbívoros y también el suelo. La bacteria produce toxinas tetanosespasmina (neurotoxina), hemolisina (tetanolisina) y toxina no espasmogénica de acción periférica que son responsables del cuadro clínico observado en los animales. Esta también se encuentra en el intestino de los equinos, siendo considerada una bacteria natural de esos animales. Se atendió un muar, macho de 12 anos de edad, peso de 250 kg, en el examen clínico el animal presentaba andar en forma de caballete, caldo en bandera, cuando se desplazó la cabeza con la protrusión del tercer párpado, se examinaron los cascos, así como todo el cuerpo del animal y no se encontró ninguna lesión, sobre la base del examen físico del animal, así como los signos clínicos presentados el diagnóstico de tétano se estableció. El tratamiento preconizado fue inmediatamente instituido con suero anti-tetánico por vía intravenosa, terapia con antibióticos y la terapia con tranquilizantes. Después de diez días del inicio del tratamiento, el animal presentaba disminución de la sintomatología de la enfermedad, retornando todos los parámetros a la normalidad, y quedando en observación por otros 10 días, presentando cura clínica.

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Tétano é uma doença infecciosa causada por toxinas do Clostridium tetani, um bacilo Gram-positivo, esporulado, anaeróbico obrigatório, que habita normalmente o trato gastrointestinal de herbívoros e também o solo. A bactéria produz as toxinas tetanosespasmina (neurotoxina), hemolisina (tetanolisina) e toxina não-espasmogênica de ação periférica que são responsáveis pelo quadro clínico observado nos animais. Essa também é encontrada no intestino dos equinos, sendo considerada uma bactéria natural desses animais. Foi atendido um muar, macho de 12 anos de idade, peso de 250 kg, no exame clínico o animal apresentava andar em forma de cavalete, calda em bandeira, quando movimentava a cabeça apresentava protrusão da terceira pálpebra, foi examinado os cascos, bem como todo o corpo do animal e não foi encontrado nenhum ferimento, com base no exame físico do animal, bem como os sinais clínicos apresentados, o diagnóstico de tétano foi estabelecido. O tratamento preconizado foi imediato instituído com soro antitetânico por via intravenosa, terapia com antibióticos e a terapia com tranquilizantes. Após 10 dias do início do tratamento, o animal apresentava diminuição da sintomatologia da enfermidade, retornando todos os parâmetros à normalidade, e ficando em observação por mais 10 dias, apresentando cura clínica.(AU)

Tetanus is an infectious disease caused by Clostridium tetani toxins, a bacterum anaerobic Gram positive, sporulated bacillus that normally inhabits the gastrointestinal tract of herbivores and also the soil. The bacterium produces the toxins tetanosespasmin (neurotoxin), hemolysin (tetanolysin) and non-spasmo genictoxin of peripheral action that are responsible for the clinical picture observed in the animals. This is also found in the intestines of horses, being considered a natural bacterium of these animals. A mule, a 12-year-old male, weighing 250 kg, was attended at the clinical examination. The animal presented a pallet-shapedwalk, flag syringe, while the head was protruding from the third eyelid, the hoofs were examined, as well as the entire body of the animal and no injury was found, based on the physical examination of the animal as well as the clinical signs presented the diagnosis of tetanus was established. The recommended treatment was immediately instituted with intravenous anti-tetanus serum, antibiotictherapy and tranquilizer therapy. Ten days after the start of treatment, the animal presented decrease of the symptomatology of the disease, returning all parameters to normality, and remaining in observation for another 10 days, presenting clinical cure.(AU)

Tétano es una enfermedad infecciosa causada por toxinas del Clostridium tetani, un bacilo Gram-positivo, esporulado, anaeróbico obligatorio, que habita normalmente el tracto gastrointestinal de herbívoros y también el suelo. La bacteria produce toxinas tetanosespasmina (neurotoxina), hemolisina (tetanolisina) y toxina no espasmogénica de acción periférica que son responsables del cuadro clínico observado en los animales. Esta también se encuentra en el intestino de los equinos, siendo considerada una bacteria natural de esos animales. Se atendió un muar, macho de 12 anos de edad, peso de 250 kg, en el examen clínico el animal presentaba andar en forma de caballete, caldo en bandera, cuando se desplazó la cabeza con la protrusión del tercer párpado, se examinaron los cascos, así como todo el cuerpo del animal y no se encontró ninguna lesión, sobre la base del examen físico del animal, así como los signos clínicos presentados el diagnóstico de tétano se estableció. El tratamiento preconizado fue inmediatamente instituido con suero anti-tetánico por vía intravenosa, terapia con antibióticos y la terapia con tranquilizantes. Después de diez días del inicio del tratamiento, el animal presentaba disminución de la sintomatología de la enfermedad, retornando todos los parámetros a la normalidad, y quedando en observación por otros 10 días, presentando cura clínica.(AU)

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Chemically inactivated tetanus toxin (tetanus toxoid, TT), purified from cultures of a virulent Clostridium tetani strain, is the active pharmaceutical ingredient of anti-tetanus vaccines. Culture clarification for TT production and is usually performed by filtration-based techniques. Final clarification of the culture supernatant is achieved by passage through 0.2 µm pore size filtering membranes. Large particles removal (primary clarification) before final filtration (secondary clarification) reduces costs of the overall clarification process. With this aim, chitosan-induced particle aggregation was assessed as an alternative for primary clarification. Three chitosan variants were tested with similar results. Optimal clarification of culture supernatant was achieved by the addition of 8 mg chitosan per l of culture. Extrapolation analysis of filter sizing results indicate that 100 l of chitosan-treated supernatant can be finally filtered with a 0.6 m2 normal filtration cartridge of 0.45 + 0.2 µm pore size. The clarified material is compatible with current standard downstream processing techniques for TT purification. Thus, chitosan-induced particle aggregation is a suitable operation for primary clarification.

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Polyvalent clostridial vaccines, composed of a complex mixture of toxoids from up to 9 different species, are highly effective in controlling clostridial diseases in cattle and sheep. Commercially available vaccines usually state that in normal field conditions two doses administered 4 to 6 weeks apart elicit protective antibody levels that will last for one year. However, studies on the development and duration of the antibody response against the different Clostridium species in target animals are scarce and only partial. Evaluating the temporal evolution of the antibody responses upon vaccination in target species is relevant to understand the bases of protective immunity induced by these vaccines and to develop new optimized vaccines. Here, we assessed the antibody response in sheep against each Clostridium component of two different 9-valent Clostridial vaccines over the period of one year. One vaccine was a commercially available vaccine and the other was an experimental vaccine prepared by us with the same antigens that we used to set up a specific ELISA for each Clostridium species. Both vaccines showed similar results, irrespectively of the origin of the antigens used for the ELISAs, with antibody titers that peaked at day 36 after vaccination and large inter individual variations in the magnitude of the response. Antibody titers were maintained up to 90 days and then markedly decreased, becoming even undetectable in some animals 6 months after vaccination. Given that the current scheme of yearly revaccination has largely shown to be effective at controlling the burden of disease, our results strongly suggest that circulating antibody levels cannot completely explain the protective immunity elicited by these vaccines, and prompt for further studies into the correlates of protection of clostridial vaccines.

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