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Clostridium perfringens α-Toxin and NetB Toxin Antibodies and Their PossibleRole in Protection Against Necrotic Enteritis and Gangrenous Dermatitis inBroiler ChickensK. W. Lee, H. S. Lillehoj, M. S. Park, S. I. Jang, G. D. Ritter, Y. H. Hong, W. Jeong, H. Y. Jeoung, D. J.An, and E. P. LillehojSource: Avian Diseases Digest, 7(1):e52-e53. 2012.Published By: American Association of Avian PathologistsDOI: http://dx.doi.org/10.1637/10029-84711-DIGEST.1URL: http://www.bioone.org/doi/full/10.1637/10029-84711-DIGEST.1

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Clostridium perfringens a-Toxin and NetB Toxin Antibodies and Their Possible Role in Protection Against NecroticEnteritis and Gangrenous Dermatitis in Broiler Chickens

K.W. Lee, H.S. Lillehoj, M.S. Park, S.I. Jang, G.D. Ritter, Y.H. Hong, W. Jeong, H.Y. Jeoung, D.J. An, and E.P. LillehojContact Address: Hyun.Lillehoj@ARS.USDA.GOV

Important FindingsNecrotic enteritis (NE) and gangrenous dermatitis (GD) are

important infectious diseases of poultry. Although NE and GDshare a common pathogen, Clostridium perfringens, they differ inother important aspects such as clinical signs, pathologic symptoms,and age of onset. The primary virulence factors of C. perfringens areits four major toxins (a, b, e, i) and the newly described NE B-like(NetB) toxin. The results of this study showed that the levels ofantibodies against both toxins were significantly higher in apparentlyhealthy chickens compared to birds with clinical signs of NE or GD,suggesting that these anti-toxin antibodies may play a role inprotection against NE and GD.

Significance of FindingsThe results of the current study highlight the importance C.

perfringens toxin antibodies in protection against bacterial infectionand raise the possibility of increasing the resistance of chickens to NEor GD by enhancing anti-toxin antibody levels.

Additional InformationNE is an acute or chronic enterotoxemia in chickens, turkeys, and

ducks caused by C. perfringens, a gram-positive, anaerobic, spore-forming bacterium, and characterized by a fibronecrotic enteritis ofthe small intestine. GD is caused by C. perfringens and, to a lesserextent C. septicum, resulting in red to black patches of moist,gangrenous skin lesions over the breast, abdomen, wing tips, or

thighs with feather loss and sloughing of the epidermis. In addition totheir unique clinical signs and symptoms, NE generally occurs at 17–18 days of age whereas GD develops at 4–7 wk. Combined with C.perfringens, co-infection with immunosuppressive viruses, reuse oflitter, high stocking density, and high viscosity diets are othercommon risks factors that have been proposed to contribute to bothdiseases.

An increasing incidence of NE in Europe subsequent to thediscontinued use of in-feed antibiotic growth promoters has beenreported. Use of antibiotic ionophores (e.g., salinomycin), however,is still permitted to control avian coccidiosis resulting from infectionby Eimeria protozoa. Some coccidiostats also possess anti-bacterialactivity against clostridia and can prevent NE. Ironically, however,GD is not uncommon in commercial poultry feeding programscontaining ionophore coccidiostats. In addition, Ritter first reportedthat NE-afflicted flocks were protected from later GD outbreaks andspeculated that early exposure to C. perfringens may induce animmune response to clostridial toxins that prevented later infections.Unlike mammals, however, the presence of antibodies against C.perfringens toxins has not been correlated with protection againstbacterial infection in chickens. In this study, researchers provideevidence that healthy chickens raised on farms with NE- or GD-afflicted chickens have higher titers of anti-toxin antibodiescompared with their diseased cohorts.

For this purpose, the researchers developed an enzyme-linkedimmunosorbent assay (ELISA) using the recently developed C.perfringens recombinant toxin proteins a-toxin and NetB toxin. TheELISA system was validated using serum samples from NE birds thatwere induced by dual challenge with Eimeria maxima and C.perfringens. Next, this ELISA was applied to field NE- or GD-afflicted chickens to monitor their serum antibody status.

Anticuerpos para las Toxinas a-Toxina y Toxina RedB de Clostridium perfringens y Su Posible Papel en la Proteccioncontra la Enteritis Necrotica y Dermatitis Gangrenosa en Pollos de Engorde

K.W. Lee, H.S. Lillehoj, M.S. Park, S.I. Jang, G.D. Ritter, Y.H. Hong, W. Jeong, H.Y. Jeoung, D.J. An, y E.P. LillehojDireccion para contactar: Hyun.Lillehoj@ARS.USDA.GOV

Hallazgos ImportantesLa enteritis necrotica (EN) y la dermatitis gangrenosa (DG) son

importantes las enfermedades infecciosas de las aves de corral. Apesar que EN y DG comparten un patogeno comun, Clostridiumperfringens, difieren en otros aspectos importantes tales como lossignos clınicos, sıntomas patologicos, y la edad de inicio. Losprincipales factores de virulencia de C. perfringens son sus cuatroprincipales toxinas (a, b, e, i) y el recientemente descrito B-NE como(RedB) toxina. Los resultados de este estudio mostraron que losniveles de anticuerpos contra las toxinas fueron significativamentemayores en pollos aparentemente sanos, en comparacion con las aves

con signos clınicos de EN o DG, lo que sugiere que estos anticuerposanti-toxina puede jugar un papel en la proteccion contra la EN y DG.

Relevancia de los HallazgosLos resultados de este estudio destacan la importancia de

anticuerpos de toxinas de C. perfringens en la proteccion contra lainfeccion bacteriana y la posibilidad de aumentar la resistencia de lospollos a EN o GD mediante la mejora de los niveles de anticuerpos dela anti-toxina.

Informacion AdicionalLa EN es una enterotoxemia aguda o cronica en los pollos, pavos y

patos causada por C. perfringens, una bacteria gram-positiva, unabacteria anaerobica, formadora de esporas, y se caracteriza por una

Copyright E 2012, American Association of Avian Pathologists, Inc. 1933-5334 online

Copyright E 2012, American Association of Avian Pathologists, Inc. 1933-5334 online

Summarized from Avian Diseases, Vol. 56, No. 1, 2012, Pages 230–233

enteritis fibronecrotica del intestino delgado. DG es causada por C.perfringens y, en menor medida, C. septicum, lo que resulta enlesiones humedas de rojas a negras, la piel gangrenada sobre el pecho,abdomen, extremidades de las alas, o en los muslos con la perdida delas plumas y el desprendimiento de la epidermis. Ademas de sussignos y sıntomas clınicos unicos, la EN se produce generalmente alos 17–18 dıas de edad, mientras que DG se desarrolla en los pollosde 4–7 semanas. En combinacion con C. perfringens, la co-infeccioncon virus inmunosupresores, la reutilizacion de la basura, la densidadde poblacion alta, y las dietas de alta viscosidad son otros factores deriesgo comunes que se han propuesto para contribuir a ambasenfermedades.

Se ha reportado una creciente incidencia de EN en Europa consubsecuente a la suspension en el uso de antibioticos promotores decrecimiento en la alimentacion. El uso de antibioticos ionoforos (porejemplo, la salinomicina), sin embargo, es aun permitido en el controlde la coccidiosis aviar consecuencia de la infeccion por protozoosEimeria. Algunos coccidiostaticos tambien poseen actividadantibacteriana contra los clostridios y puede prevenir la EN.Ironicamente, sin embargo, la DG no es comun en los programas

comerciales de alimentacion de aves de corral que contienencoccidiostaticos ionoforos. Ademas, Ritter informo por primera vezque las parvadas afectadas por EN fueron protegidas de los brotestardıos de DG y especularon que la exposicion temprana a C.perfringens puede inducir una respuesta inmune a las toxinasclostridiales que impidio infecciones posteriores. A diferencia de losmamıferos, sin embargo, la presencia de anticuerpos contra las toxinasde C. perfringens no se ha correlacionado con la proteccion contra lainfeccion bacteriana en los pollos. En este estudio, los investigadoresproporcionan pruebas de que los pollos sanos criados en granjas conpollos afectados con EN-DG tienen tıtulos mayores de anticuerposanti-toxina en comparacion con sus cohortes de enfermos.

Para ello, los investigadores desarrollaron un ensayo deinmunoabsorcion ligado a enzimas (ELISA), utilizando lasrecientemente desarrolladas proteınas recombinantes de toxinas a yRedB de C. perfringens. El sistema ELISA fue validado utilizandomuestras de suero de las aves con EN que fueron inducidas por dobledesafıo con Eimeria maxima y Clostridium perfringens. Acontinuacion, este ELISA se aplico al campo en pollos afectadoscon EN-DG para controlar su estado de anticuerpos en suero.

Toxina especıfica de C. perfringens y anticuerpos en aves EN y DG