Mechanisms of Action and Cell Death Associated with Clostridium perfringens Toxins.

Clostridium perfringens uses its large arsenal of protein toxins to produce histotoxic, neurologic and intestinal infections in humans and animals. The major toxins involved in diseases are alpha (CPA), beta (CPB), epsilon (ETX), iota (ITX), enterotoxin (CPE), and necrotic B-like (NetB) toxins. CPA is the main virulence factor involved in gas gangrene in humans, whereas its role in animal diseases is limited and controversial. CPB is responsible for necrotizing enteritis and enterotoxemia, mostly in neonatal individuals of many animal species, including humans. ETX is the main toxin involved in enterotoxemia of sheep and goats. ITX has been implicated in cases of enteritis in rabbits and other animal species; however, its specific role in causing disease has not been proved. CPE is responsible for human food-poisoning and non-foodborne C. perfringens-mediated diarrhea. NetB is the cause of necrotic enteritis in chickens. In most cases, host⁻toxin interaction starts on the plasma membrane of target cells via specific receptors, resulting in the activation of intracellular pathways with a variety of effects, commonly including cell death. In general, the molecular mechanisms of cell death associated with C. perfringens toxins involve features of apoptosis, necrosis and/or necroptosis

Biodiversity and ecosystem services in the Frome catchment, Purbeck district, United Kingdom

A map for valuing ecosystem services in the 480 km2 Frome catchment, to investigate scenarios of change in land use, was internet crowd-sourced. Scouts mapped deer habitats in 15% of the 30 km2 Arne Parish, while 143 residents volunteered data on deer sightings in the 5-year community survey

Proteolytic processing and activation of Clostridium perfringens epsilon toxin by caprine small intestinal contents.

Epsilon toxin (ETX), a pore-forming toxin produced by type B and D strains of Clostridium perfringens, mediates severe enterotoxemia in livestock and possibly plays a role in human disease. During enterotoxemia, the nearly inactive ETX prototoxin is produced in the intestines but then must be activated by proteolytic processing. The current study sought to examine ETX prototoxin processing and activation ex vivo using the intestinal contents of a goat, a natural host species for ETX-mediated disease. First, this study showed that the prototoxin has a KEIS N-terminal sequence with a molecular mass of 33,054 Da. When the activation of ETX prototoxin ex vivo by goat small intestinal contents was assessed by SDS-PAGE, the prototoxin was processed in a stepwise fashion into an ~27-kDa band or higher-molecular-mass material that could be toxin oligomers. Purified ETX corresponding to the ~27-kDa band was cytotoxic. When it was biochemically characterized by mass spectrometry, the copresence of three ETX species, each with different C-terminal residues, was identified in the purified ~27-kDa ETX preparation. Cytotoxicity of each of the three ETX species was then demonstrated using recombinant DNA approaches. Serine protease inhibitors blocked the initial proteotoxin processing, while carboxypeptidase inhibitors blocked further processing events. Taken together, this study provides important new insights indicating that, in the intestinal lumen, serine protease (including trypsin and possibly chymotrypsin) initiates the processing of the prototoxin but other proteases, including carboxypeptidases, then process the prototoxin into multiple active and stable species. Importance: Processing and activation by intestinal proteases is a prerequisite for ETX-induced toxicity. Previous studies had characterized the activation of ETX using only arbitrarily chosen amounts of purified trypsin and/or chymotrypsin. Therefore, the current study examined ETX activation ex vivo by natural host intestinal contents. These analyses demonstrated that (i) ETX processing in host intestinal contents occurs in an ordered, stepwise fashion, (ii) processing of prototoxin by host intestinal contents results in higher-molecular-mass material and 3 distinct ~27-kDa ETX species, and (iii) serine proteases, such as trypsin, chymotrypsin, and other proteases, including carboxypeptidases, play a role in the activation of ETX by intestinal contents. These studies provide new insights into the activation and processing of ETX and demonstrate that this process is more complicated than previously appreciated

The VirS/VirR two-component system regulates the anaerobic cytotoxicity, intestinal pathogenicity, and enterotoxemic lethality of Clostridium perfringens type C isolate CN3685.

Clostridium perfringens vegetative cells cause both histotoxic infections (e.g., gas gangrene) and diseases originating in the intestines (e.g., hemorrhagic necrotizing enteritis or lethal enterotoxemia). Despite their medical and veterinary importance, the molecular pathogenicity of C. perfringens vegetative cells causing diseases of intestinal origin remains poorly understood. However, C. perfringens beta toxin (CPB) was recently shown to be important when vegetative cells of C. perfringens type C strain CN3685 induce hemorrhagic necrotizing enteritis and lethal enterotoxemia. Additionally, the VirS/VirR two-component regulatory system was found to control CPB production by CN3685 vegetative cells during aerobic infection of cultured enterocyte-like Caco-2 cells. Using an isogenic virR null mutant, the current study now reports that the VirS/VirR system also regulates CN3685 cytotoxicity during infection of Caco-2 cells under anaerobic conditions, as found in the intestines. More importantly, the virR mutant lost the ability to cause hemorrhagic necrotic enteritis in rabbit small intestinal loops. Western blot analyses demonstrated that the VirS/VirR system mediates necrotizing enteritis, at least in part, by controlling in vivo CPB production. In addition, vegetative cells of the isogenic virR null mutant were, relative to wild-type vegetative cells, strongly attenuated in their lethality in a mouse enterotoxemia model. Collectively, these results identify the first regulator of in vivo pathogenicity for C. perfringens vegetative cells causing disease originating in the complex intestinal environment. Since VirS/VirR also mediates histotoxic infections, this two-component regulatory system now assumes a global role in regulating a spectrum of infections caused by C. perfringens vegetative cells

First report of caprine abortions due to Chlamydia abortus in Argentina.

Infectious abortions of goats in Argentina are mainly associated with brucellosis and toxoplasmosis. In this paper, we describe an abortion outbreak in goats caused by Chlamydia abortus. Seventy out of 400 goats aborted. Placental smears stained with modified Ziehl-Neelsen stain showed many chlamydia-like bodies within trophoblasts. One stillborn fetus was necropsied and the placenta was examined. No gross lesions were seen in the fetus, but the inter-cotyledonary areas of the placenta were thickened and covered by fibrino-suppurative exudate. The most consistent microscopic finding was found in the placenta and consisted of fibrinoid necrotic vasculitis, with mixed inflammatory infiltration in the tunica media. Immunohistochemistry of the placenta was positive for Chlamydia spp. The results of polymerase chain reaction targeting 23S rRNA gene performed on placenta were positive for Chlamydia spp. An analysis of 417 amplified nucleotide sequences revealed 99% identity to those of C. abortus pm225 (GenBank AJ005617) and pm112 (GenBank AJ005613) isolates. To the best of our knowledge, this is the first report of abortion associated with C. abortus in Argentina

Chronic inflammatory lung lesions in rabbits free of known respiratory pathogens

Chronic inflammatory lung lesions were studied in 58 purpose bred, clinically healthy rabbits that were found negative for Bordetella branchiseptica, Pasleurel/a multocida, Toxoplasma gondii, Encephalitozoon cuniculi, and lung worms. The changes found consisted of focal chronic interstitial pneumonia and chronic bronchitis and bronchiolitis, which were frequently associated with perivascular infiltrations of mononuclear leukocytes and vasculitis involving small lung vessels. The etiology of these alterations remains unknown

Scale-dependent effects of density and habitat on foal survival

Identifying the most appropriate scale to study factors influencing life history is important to evolutionary ecology and wildlife management. For example, the scale at which density is assessed and explains variation in survival can affect how biologists observe and interpret population dynamics, which can influence plans for managing populations. Feral horses (Equus ferus caballus) contrast with most ungulates by exhibiting a mating system characterized by female-defense polygyny with persistent, non-territorial breeding groups (bands) and female-biased initial (natal) and subsequent (breeding) dispersal. We predicted that for horses, offspring movements coupled with female-biased breeding dispersal would increase the scale at which density best related to juvenile survival compared to species with greater female philopatry. From 2008 to 2013, we censused the population of feral horses on Sable Island, Canada. We annually computed individual-specific local densities for 442 foals (horses/km2 in radii of 2,000 m, 4,000 m, and 8,000 m fixed to a band’s centroid of movements) and whole-island (total) population density, group (band) size, and local access to surface freshwater, which affected movement patterns and selection of vegetation by females. The population of feral horses increased from 380 in 2008 to 559 in 2013. Overwinter survival of foals averaged 82.8%. Island-wide density was the most important predictor of foal mortality and was negatively associated with survival, with a lesser negative effect from local density. Increased access to surface freshwater (ponds) was an important predictor of foal survival but only at certain scales. Our study emphasizes the relevance of a multi-scale approach when analyzing the response of fitness components to changes in habitat and population processes, which may be influenced by the particular social organization of the species

A study of lung lesions in asymptomatic rabbits naturally infected with B. bronchiseptica

A histological study of lungs performed in 40 rabbits carrying a naturally acquired subclinical B. bronchiseptica infection revealed chronic inflammatory pulmonary lesions in 38 rabbits, B. bronchiseptica was the only agent consistently isolated from lungs. Pasteurella spp. was not isolated from any animal. B. bronchiseptica infected rabbits frequently displayed changes characterized by a simultaneous occurrence of focal chronic interstitial pneumonia, vascular and perivascular infiltration of monocytes and lymphoid cells, and inflammation ofbronchi and bronchioli

Distribución y aspectos poblacionales del lobo Ibérico en la provincia de Ourense

Para determinar el área de distribución del lobo en la provincia de Ourense (noroeste de España) se realizaron prospecciones de campo consistentes en itinerarios de muestreo para localizar indicios y se enviaron cuestionarios a los Agentes del Servicio de Conservación de la Naturaleza de la Xunta de Galicia y a las Sociedades de Cazadores del área de estudio. Los resultados se registraron sobre cuadrículas U.T.M. 10x10 Km. Se constata presencia del lobo en el 79,2% de las cuadrículas estudiadas, estimando el área de distribución en la provincia en 6.400 km2. Para la localización de los grupos familiares inicialmente se prospectó el territorio a nivel de cuadrículas U.T.M. 10x10 Km y en función de los resultados y la información recibida se eligieron determinadas zonas para realizar estaciones de escucha y espera. Determinamos durante el periodo de estudio la existencia de 25 grupos familiares. La densidad estimada en la provincia es de 2,10 – 3,28 lobos /100 km2. De los casos de mortalidad conocidos entre enero de 1999 y abril de 2002 (n=37), un elevado porcentaje se debió a atropellos (70,27%) principalmente en dos vías concretas y un 8,10% correspondió a envenenamientos. Actualmente la densidad del lobo en Ourense es superior a la obtenida en provincias limítrofes, pero amenazas de origen antrópico tanto directas (veneno, persecución ilegal...) como indirectas (incendios forestales, grandes infraestructuras viales, parques eólicos...) pueden estar condicionando la existencia de grupos familiares e incluso la presencia de la especie en determinadas zonas

Diagnóstico de enfermedades clostridiales de los animales

Las enfermedades clostridiales son producidas por bacterias del género Clostridium, bacilos grampositivos, anaerobios y esporulados. La gran mayoría de los animales domésticos y silvestres son susceptibles a una o más de las enfermedades clostridiales descriptas en este resumen. Las enfermedades clostridiales de los animales se clasifican en entéricas, histotóxicas y neurotóxicas.Trabajo publicado en Cagliada, Maria del Pilar Lilia y Galosi, Cecilia Mónica (comps.). I Congreso de Microbiología Veterinaria. Libro de resúmenes. La Plata: Facultad de Ciencias Veterinarias, 2021.Facultad de Ciencias Veterinaria