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Necrotizing lesions in the intestine, gizzard, and liver in captive capercaillies (Tetrao urogallus) associated with Clostridium perfringens.

During the period from 1982 to 1991, 863 captive and 32 wild capercaillies (Tetrao urogallus) were necropsied. The most common cause of death in captive capercaillies was necrotizing enteritis, diagnosed in 110 (13%) birds. Of these, 31 (28%) birds also had necrotizing lesions in the liver. Necrotizing gastritis occurred in 29 birds, two of which had concurrent necrotizing enteritis. In the capercaillies with necrotizing enteritis, Clostridium perfringens type A was isolated more frequently and in larger numbers than in birds which died from other causes. Thus, Clostridium perfringens type A may be of etiological importance in necrotizing enteritis. Necrotizing enteritis was not diagnosed in wild capercaillies.

Animals↗

Isolation of Clostridium tertium from a striped dolphin (Stenella coeruleoalba) in the Adriatic Sea.

Bacteriologic examination of an abscess found between blubber and musculature of a striped dolphin (Stenella coeruleoalba) found dead on 30 March 2002 in the Adriatic Sea, Croatia, was performed and an aerotolerant, nontoxogenic Clostridium tertium was isolated in pure culture. National Collections of Industrial Food and Marine Bacteria (NCIMB Ltd., Aberdeen, Scotland, UK) confirmed the results. Sequencing results showed it to be C. tertium with 100% similarity. The strain was named Clostridium tertium strain Zagreb, acceded to the culture collection and assigned the accession number NCIMB 13970. This is the first record of C. tertium in marine mammals.

Animals↗

Evaluation of methods to diagnose Clostridium perfringens-associated diarrhea in dogs.

OBJECTIVE: To assess the prevalence of Clostridium perfringens enterotoxin in feces of dogs with and without diarrhea, and to compare the use of microbial cultures from fecal specimens and evaluation of stained fecal smears for endospores with the presence of enterotoxin as tools for diagnosing C perfringens-associated diarrhea. DESIGN: Prospective study. ANIMALS: 144 dogs representing hospitalized dogs with (n = 41) or without (50) diarrhea, and clinically normal dogs treated as outpatients (53). PROCEDURE: Fresh fecal specimens from all dogs were examined as Gram-stained fecal smears to determine numbers of Gram-positive spore-forming rods/100x objective field. Enterotoxin was assayed directly by use of a reverse passive latex agglutination assay. Fecal specimens were plated directly to prereduced egg yolk agar plates and incubated overnight at 37 C in an anaerobic chamber. At 24 hours, up to 3 lecithinase-positive colonies were subcultured to Brucella blood agar to evaluate for double zone hemolysis. Colonies with double zone hemolysis were tested for aerotolerance and Gram-stained. RESULTS: A significant difference was not detected among groups with respect to the presence of C perfringens as determined by culture, the presence of endospores, and the reaction patterns of fecal enterotoxin assays. An association was not found between number of endospores and the presence of fecal enterotoxin. CLINICAL IMPLICATIONS: The presence of C perfringens enterotoxin in feces of dogs, as detected by the latex agglutination assay used in this study, correlates poorly with the number of fecal endospores, regardless of the dog's clinical status.

Animals↗

Human diseases caused by exotoxins produced by anaerobes and their rapid detection.

Major human diseases caused by exotoxins produced by anaerobes include botulisms, tetanus, foodborne illness caused by enterotoxin-producing Clostridium perfringens, and diarrhea/colitis caused by toxigenic Clostridium difficile. Recently, enterotoxigenic Bacteroides fragilis (ETBF) has been recognized, that may be related to childhood diarrheal disease. Detection test of botulinal neurotoxin is hardly performed at clinical laboratories since the most reliable means of detection and identification of botulinal toxin is by using mouse toxicity and neutralization tests. Clinical laboratories should request the tests to a reference laboratory. Since tetanus is easily diagnosed clinically on the basis of its unique, recognizable sings, the bacteriological tests is not usually requested. C. perfringens foodborne illness can be confirmed by testing stool specimens or the suspect food(s) for enterotoxin by the reversed passive latex agglutination test or counting > 10 5 C. perfringens per g of suspected food or > 10 6 C. perfringens spores per g of stool. Diagnosis of C. difficile-associated diarrhea/colitis is confirmed by detection of toxins A or B of C. difficile and/or recovery of toxigenic C. difficile. Isolation of C. difficile strains or detection C. difficile-speciffic antigen from stool specimens is less diagnostic since nontoxic or toxin A positive-toxin B negative strains are prevalent in Japan. Reliable laboratory tests for ETBF-associated childhood diarrhea are not established yet although ETBF can be proved by polymerase chain reaction for detection of the enterotoxin gene.

Animals↗

Prevalence of Clostridium perfringens enterotoxin and Clostridium difficile toxin A in feces of horses with diarrhea and colic.

OBJECTIVE: To determine prevalence of clostridial enterotoxins in feces of horses with diarrhea and colic, and to determine whether an association exists between detection of clostridial enterotoxins in feces and development of diarrhea as a complication of colic. DESIGN: Prospective case series and case-control study. ANIMALS: 174 horses with diarrhea, colic, or problems not related to the gastrointestinal tract. PROCEDURE: Horses were assigned to 1 of 4 groups: colic with diarrhea (group 1; n = 30); colic without diarrhea (group 2; 30); diarrhea without colic (group 3; 57); and control (group 4; 57). Feces were evaluated by use of ELISA to detect Clostridium perfringens enterotoxin (CPE) and C difficile toxin A (TOXA). Frequency of detection of CPE or TOXA in groups 1 and 3 was compared with that in groups 2 and 4, respectively. RESULTS: Prevalence of enteric clostridiosis in horses in group 3 was 25%. Clostridium perfringens enterotoxin was detected in 9 of 57 (16%), TOXA in 8 of 57 (14%), and both toxins in 3 of 57 (5%) fecal samples collected from these horses. Neither toxin was detected in feces of the age-matched horses in group 4. Clostridial enterotoxins were detected in feces of 7 of 60 (12%) horses with colic (groups 1 and 2), however, a significant association was not found between detection of enterotoxins in feces and development of diarrhea as a complication of colic. CONCLUSIONS AND CLINICAL RELEVANCE: Clostridia are important etiologic agents of diarrhea in horses. Additionally, changes in intestinal flora of horses with colic may allow for proliferation of clostridia and elaboration of enterotoxins regardless of whether diarrhea develops.

Animals↗

Cecal carriage of Clostridium perfringens in broiler chickens given Mucosal Starter Culture.

Day-of-hatch broiler chicks housed in isolation units were each given, by oral gavage, 0.1 ml of Mucosal Starter Culture (MSC) or saline control. Each of the treated and control chicks was subsequently given a composite culture of three strains of bacitracin-resistant Clostridium perfringens (Cp) previously isolated from chickens with symptoms of necrotic enteritis. Some chicks were maintained on a corn-based diet provided ad libitum. Others were given the feed supplemented with 50% rye (a predisposing factor for necrotic enteritis). At 7, 14, and 21 days after receiving Cp, chicks were euthanatized, and cecal contents were diluted and plated on selective agar containing bacitracin. For chicks on corn feed, Cp numbers were similar in control birds and birds given MSC in three of four trials. In two of the trials that demonstrated no effect of MSC on Cp numbers, enterotoxin presence was determined. The number of birds with detectable Cp enterotoxin in their small intestine and the mean toxin levels were lower in the MSC-treated birds. In a fourth trial with birds on corn-based feed, mean Cp numbers and the number of Cp-positive birds were lower in the MSC-treated birds. For the two trials involving chickens on rye-supplemented feed, Cp numbers and the percentage of Cp-positive birds were significantly reduced in MSC-treated birds compared with control birds. Enterotoxin in birds receiving the 50% rye diet was at low levels or not detected in control and MSC-treated birds. Results suggest that MSC may reduce intestinal proliferation of Cp, a causative agent of necrotic enteritis in poultry and of foodborne disease in humans.

Animal Feed↗

Development of monoclonal antibodies suitable for use in antigen quantification potency tests for clostridial veterinary vaccines.

The quality control testing of clostridial veterinary vaccines currently requires large numbers of animals. Alternative in vitro test methods are being investigated by researchers in industry and by regulatory authorities in many countries. Monoclonal antibodies that neutralize Clostridium perfringens alpha toxin, C. perfringens beta toxin, C. perfringens epsilon toxin, and C. sordellii lethal toxin as well as a monoclonal antibody directed against C. chauvoei flagellar antigen have been developed by the Center for Veterinary Biologics-Laboratory for use in antigen quantification assays. A proposal to create an international standard collection of clostridial-specific monoclonal antibodies is made.

Animal Welfare↗

The impact of various browse feeds with different tannin content on the fecal shedding of Clostridium perfringens in West African dwarf sheep.

In 1994 and 1995 leaves from eight browse feeds, containing tannins in different amounts (BF), were fed to West African Dwarf Sheep in Benin to evaluate their impact on Clostridium perfringens in the intestinal tract. An inhibitory impact of various BF on the growth of C. perfringens was assessed in in-vitro assays before, and thus a potential use of these leaves as a preventive diet against C. perfringens enterotoxemia in small ruminants was assumed. Surprisingly, an inhibitory impact of the BF on the shedding of C. perfringens in the feces of West African Dwarf Sheep could not be shown in seven of the eight BF examined. However, the pattern of inhibition of unlike C. perfringens toxovars may differ and a selective inhibitory impact of the BF Dialium guineense on C. perfringens toxovar D may be assumed.

Africa, Western↗