Anaerobic bacterial infections and response to treatment in dogs and cats: 36 cases (1983-1985)

Anaerobic bacteria have been increasingly implicated as important pathogens in animals. To determine the prevalence of anaerobic bacterial infection, the results of anaerobic bacteriologic culture of 599 specimens obtained from dogs and cats hospitalized at the Colorado State University Veterinary Teaching Hospital were reviewed. Obligate anaerobic bacteria were isolated from 35% of properly submitted specimens; Bacteroides spp and Fusobacterium spp were the organisms most commonly isolated. Infections most often containing anaerobes were abscesses, pleuropulmonary infections, and abdominal infections. A complication rate of 28% was observed with anaerobic bacterial infections; failure to initially treat with an effective antibiotic increased the rate of recurrence of infection.     

Aerobic and anaerobic bacterial isolates from horses with pneumonia or pleuropneumonia and antimicrobial susceptibility patterns of the aerobes

Frequency of aerobic and anaerobic isolates in 327 aspirates and in 123 pleural fluid samples from 327 horses with pneumonia or pleuropneumonia and antimicrobial susceptibility patterns of the aerobes were reported. Of the 327 horses, 75% survived, 20% were euthanatized, and 5% died. Tracheobronchial aspirates or pleural fluid specimens from 25 of the horses did not yield growth. Of the remaining 302 horses, 221 had only aerobic organisms isolated, whereas only anaerobes were isolated from 6 of the 302 horses. The remaining 75 horses had mixed aerobic and anaerobic bacterial infections. The survival rates for horses with aerobic only isolates was twice that of horses with anaerobic isolates. The aerobic bacteria most frequently isolated were beta-Streptococcus spp, Pasteurella spp, Escherichia coli, and Enterobacter spp. The anaerobic species most frequently isolated were Bacteroides spp and Clostridium spp.     

Effects of dietary fructooligosaccharide on selected bacterial populations in feces of dogs

OBJECTIVE: To evaluate fecal concentrations of selected genera of colonic bacteria in healthy dogs, and to investigate effects of dietary fructooligosaccharides (FOS) on those bacterial populations. ANIMALS: 6 healthy adult Beagles. PROCEDURE: Dogs were randomly assigned to 2 groups of 3 and fed an unsupplemented diet for 370 days. After 88 days, fecal samples were collected. Another fecal sample was collected from each dog 282 days later. Group A then received a diet supplemented with FOS, and group B continued to receive the unsupplemented diet. Twenty-eight to 29 days later, fecal samples were collected. Diets were switched between groups, and fecal samples were collected 31 and 87 days later. Concentrations of Bifidobacterium spp, Lactobacillus spp, Clostridium spp, Bacteroides spp, and Escherichia coli in freshly collected feces were determined. Effects of diet and time on bacterial concentrations were compared between groups. RESULTS: Bifidobacterium spp and Lactobacillus spp were inconsistently isolated from feces of dogs fed either diet. Sequence of diet significantly affected number of Bacteroides spp subsequently isolated from feces, but diet had no effect on numbers of Clostridium spp or E coli. CONCLUSIONS AND CLINICAL RELEVANCE: Some genera of bacteria (eg, Bifidobacterium) believed to be common components of colonic microflora may be only sporadically isolated from feces of healthy dogs. This deviation from expected fecal flora may have implications for the effectiveness of supplementing diets with prebiotics.     

Enteric Clostridium perfringens infection associated with parvoviral enteritis in dogs: 74 cases (1987-1990).

Parvovirus infection was confirmed by fluorescent antibody staining of or viral isolation from specimens of small intestine in 181 (17%) of 1,110 dogs necropsied between July 1, 1987 and Dec 31, 1990. Clostridium perfringens was isolated from 74 (69%) of 108 dogs with parvovirus infection from which specimens of jejunum also had been obtained for culture of anaerobic bacteria. Gram-positive bacilli in association with focal to diffuse necrosis of the superficial portions of the villi were observed in histologic sections of specimens of small intestine from 56 (98%) of 57 dogs from which parvovirus and C perfringens had been identified. These findings indicate that C perfringens frequently proliferates in dogs with parvovirus infection.     

The roles of Clostridium difficile and enterotoxigenic Clostridium perfringens in diarrhea in dogs

In this prospective study, feces of dogs with diarrhea were compared with feces of normal dogs for the presence of Clostridium difficile, C difficile toxins A and B, C perfringens, and C perfingens enterotoxin (CPE). C difficile toxins A, B, or both were present in feces of 18 of 87 (21%) dogs with diarrhea and 4 of 55 (7%) normal dogs (P = 0.03), whereas CPE was present in the feces of 24 of 87 (28%) dogs with diarrhea and 3 of 55 (5%) normal dogs (P = 0.01). C difficile was isolated from 2 of 87 (2%) dogs with diarrhea but was not isolated from the feces of 55 normal dogs, possibly because of poor survival of the organism in fecal samples. C perfringens was isolated from the feces of 23 of 24 (96%) CPE-positive dogs with diarrhea, 52 of 63 (83%) CPE-negative dogs with diarrhea, and 39 of 55 (71%) CPE-negative dogs with normal feces. No correlation was found between C perfringens spore number and the presence of CPE.     

Evaluation of a routine diagnostic fecal panel for dogs with diarrhea

OBJECTIVES: To assess the diagnostic yield of a routine fecal panel and determine whether Clostridium perfringens or C difficile toxin production is associated with acute hemorrhagic diarrheal syndrome (AHDS) in dogs. DESIGN: Case-control study. ANIMALS: 260 dogs with diarrhea and 177 dogs with normal feces. PROCEDURE: Medical records were reviewed for results of culture for C difficile, Campylobacterspp, and Salmonella spp; C perfringens fecal enterotoxin (CPE) assay via ELISA or reverse passive latex agglutination (RPLA) assay; fecal endospore enumeration; C difficile toxin A assay; and parasite evaluation. RESULTS: Prevalence of CPE in dogs with diarrhea was 22/154 (14.3%) via ELISA and 47/104 (45.2%) via RPLA assay, versus 9/74 (12%) via ELISA and 26/103 (25%) via RPLA assay in control dogs. Prevalence of C difficile was 47/260 (18%) in dogs with diarrhea and 41/74 (55%) in control dogs. Prevalence of C difficile toxin A was 26/254 (10.2%) in dogs with diarrhea and 0/74 in control dogs. Diagnosis of AHDS was made in 27 dogs; 8 had positive results for CPE, 7 had positive results for toxin A, and 1 had positive results for both toxins. Campylobacter spp were isolated from 13 of 260 (5%) dogs with diarrhea and 21 of 74 (28.4%) control dogs. Salmonella spp were isolated from 3 (1.2%) dogs with diarrhea. CONCLUSIONS AND CLINICAL RELEVANCE: Diagnostic value of a fecal panel in dogs with diarrhea appears to below.     

Comparison of bacteriologic culture of blood and necropsy specimens for determining the cause of foal septicemia: 47 cases (1978-1987)

Diagnosis of bacterial septicemia was confirmed by results of bacteriologic culture of antemortem blood samples and/or necropsy specimens obtained from 47 foals up to 8 days old. Gram-negative bacteria were isolated from all 47 foals, and mixed infections with more than one organism were involved in 26 (55%). Escherichia coli, Actinobacillus spp, and Klebsiella pneumoniae were the most frequent isolates, infecting 55, 34, and 23% of foals, respectively. Gram-positive bacteria and anaerobic bacteria were isolated only from foals with mixed infections with gram-negative organisms. Clostridium perfringens was the only anaerobe isolated. In 38 (81%) of 47 foals with confirmed septicemia, blood samples were culture-positive. Thirty-two septicemic foals subsequently died, allowing a comparison to be made between the species of bacteria isolated by culture of blood with those recovered by culture of internal organs at necropsy. Blood failed to yield any gram-negative organisms in 12 (37.5%) of 32 foals from which a gram-negative pathogen was isolated at necropsy. Forty-three percent of the gram-negative bacteria, including 59% of the E coli, and 10% of the gram-positive bacteria found in septicemic foals at necropsy were not detected earlier by results of bacteriologic culture of blood.     

Genotypic and phenotypic characterization of Clostridium perfringens and Clostridium difficile in diarrheic and healthy dogs

The objectives of this study were to examine the potential roles of Clostridium difficile and enterotoxigenic Clostridium perfringens in diarrhea in dogs by comparison of isolation, determination of toxin status via enzyme-linked immunosorbent assay (ELISA), and application of multiplex polymerase chain reaction (PCR). These techniques were used to evaluate fecal specimens in 132 healthy and diarrheic dogs. These dogs were prospectively evaluated by grouping them into the following 3 categories: hospitalized dogs with diarrhea (n = 32), hospitalized dogs without diarrhea (n = 42), and apparently healthy outpatient dogs without diarrhea (n = 58). All fecal specimens were cultured using selective media for C difficile, Salmonella spp., and Campylobacter spp. and selective media after heat shock for C perfringens. No significant difference was found in the isolation of C perfringens or C difficile among the 3 groups. A significant association was found between the presence of diarrhea and detection of C perfringens enterotoxin (CPE) or toxin A via ELISA for both C perfringens and C difficile, respectively. PCR performed on C difficile isolates for toxin A and toxin B genes revealed no significant differences among the 3 groups, but diarrheic dogs were significantly more likely to be positive for the enterotoxin gene of C perfringens. Based on the results of this study, the use of ELISA for detection of CPE in feces combined with the detection of enterotoxigenic fecal isolates obtained via heat shock provides the strongest evidence for the presence of C perfringens-associated diarrhea.     

Obligately anaerobic bacterial species isolated from foot-rot lesions in goats

Lesions showing clinical signs of foot-rot from 120 goats were cultured on six selective media during October 1987 to November 1988. A total of 582 strictly anaerobic microorganisms belonging to 50 different species was isolated and identified. The anaerobes most frequently isolated belonged to the following genera: Bacteroides (80%), Peptostreptococcus (63.6%), Megasphaera (40%), Fusobacterium (29.2%), Clostridium (22.5%), Propionibacterium (12.5%), Eubacterium (11.7%) and Leptotrichia (10.8%). Percentages for Acidaminococcus, Peptococcus, Tissierella, Wolinella and Veillonella were below 10%. The following species were identified in 10% or more of cases: Peptostreptococcus anaerobius (61.7%), Bacteroides buccae (51.7%), Bacteroides nodosus (42.5%), Megasphaera elsdenii (40%), Bacteroides ruminicola subsp. brevis (22.5%), Fusobacterium necrophorum (19.2%), Leptotrichia buccalis (11.7%) and Clostridium perfringens (10%). Lower percentages were obtained for the remaining species. The efficiency and selectivity of the six culture media used for isolation are discussed.     

Bacteria in enteric lesions of cattle

Thirty-nine species of bacteria were isolated from or demonstrated in the abomasal and small and large intestinal mucosa of 23 adult cattle and 41 calves and identified. The bacteria isolated were related to the gross and microscopical lesions. Campylobacters, Clostridium perfringens type A, C sordellii, Actinobacillus lignieresii, Fusobacterium necrophorum, Mycobacterium paratuberculosis, Aeromonas hydrophila and Escherichia coli were all associated with specific lesions. The relationship of other bacteria such as Bacillus licheniformis, Branhamella catarrhalis and Bacteroides vulgatus to lesions in which they were found was discussed. It was concluded that some of the bacteria could be responsible for the lesions in which they were found. However, proof of this supposition could only be obtained by experimental infections of non-immune cattle with pure cultures.     

Bacterial infection of the lower respiratory tract in 34 horses

OBJECTIVE: To investigate associations between the bacteriology and aspects of history, clinical presentation, outcome and pathology of lower respiratory tract disease of 34 horses. PROCEDURE: Detailed aerobic and anaerobic bacteriological investigations were performed on clinical specimens from horses with pneumonia, lung abscessation and necrotic pneumonia with or without pleurisy in an attempt to identify those bacteria that might contribute to the initiation and progression of infection. RESULTS: Bacteria were cultured from 33 of the 34 horses. In ten cases, only aerobic/facultatively anaerobic isolates were cultured while aerobic/facultatively anaerobic bacteria and obligately anaerobic bacteria were isolated in the other 23 cases. Moderate to large numbers of anaerobic bacteria were isolated only when the estimated duration of illness was at least five days. Bacteria were not cultured from 12 of the pleural fluid samples but were always cultured from pulmonary samples (either transtracheal aspirates from live horses or pulmonary lesions at necropsy). Streptococcus equi subsp zooepidemicus was isolated in the three cases where only one bacterial species was cultured. In the other 30 cases, multiple species were isolated. These included most often and in greatest numbers, Streptococcus equi subsp zooepidemicus, Pasteurellaceae, Escherichia coli, anaerobic cocci, Eubacterium fossor, Bacteroides tectum, Prevotella heparinolytica, Fusobacterium spp, and pigmented members of the genera Prevotella and Porphyromonas. Aerobic/facultatively anaerobic organisms were isolated from 97% of horses, while obligately anaerobic organisms were cultured from 68% of horses. CONCLUSION: There was no association between the isolation of any specific bacterium and the outcome of disease. However, obligately anaerobic bacteria (such as anaerobic cocci, Bacteroides tectum, P heparinolytica and Fusobacterium spp) and the facultatively anaerobic species Escherichia coli, were recovered more commonly from horses that died or were euthanased than from those that survived. There was an association between failure of horses to recover from pleuropneumonia and delay in diagnosis and initiation of treatment.     

The relationship between the presence of Helicobacter pylori, Clostridium perfringens type A, Campylobacter spp, or fungi and fatal abomasal ulcers in unweaned beef calves.

A case-control study involving 30 unweaned beef calves was conducted to determine whether specific species of bacteria or fungi were associated with fatal abomasal ulcer formation. Special microbiological and histological techniques were used to detect Clostridium perfringens type A, Helicobacter pylori, or Campylobacter spp. It has been speculated that these bacteria are potential ulcerogenic agents of unweaned beef calves. Calves were recruited for the study at necropsy, with those dying of either a perforating or a hemorrhagic ulcer representing the cases, and calves of a similar age dying of a disease unrelated to the abomasum representing the controls. Helicobacter pylori was not visualized in or cultured from any of the abomasal tissue samples. Clostridium perfringens type A was isolated from 78.6% of the cases and 75% of the controls. These isolates were further dichotomized into "heavy" and "light" growth; no significant association was found between ulcers and the amount of growth. A light growth of Campylobacter spp. was recovered from 3 cases and 3 controls. There was no compelling evidence to suggest that Clostridium perfringens type A, Helicobacter pylori, or Campylobacter spp. were involved in ulcer formation.     

Oral associated bacterial infection in horses: studies on the normal anaerobic flora from the pharyngeal tonsillar surface and its association with lower respiratory tract and paraoral infections

Two hundred and seventy bacterial isolates were obtained from the pharyngeal tonsillar surface of 12 normal horses and 98 obligatory anaerobic bacteria were characterised. Of these, 57 isolates belonging to 7 genera (Peptostreptococcus (1); Eubacterium (9); Clostridium (6); Veillonella (6); Megasphera (1); Bacteroides (28); Fusobacterium (6)) were identified, and 16 of these were identified to species level (P. anaerobius (1); E. fossor (9); C. villosum (1); B. fragilis (1); B. tectum (2); B. heparinolyticus (2)). Three hundred and twenty isolates were obtained from 23 samples from horses with lower respiratory tract (LRT) or paraoral (PO) bacterial infections. Of the 143 bacteria selected for detailed characterisation, obligate anaerobes accounted for 100 isolates, facultative anaerobes for 42 isolates and obligate aerobes for one isolate. Phenotypic characterisation separated 99 of the isolates into 14 genera. Among the obligately anaerobic species, Gram-positive cocci including P. anaerobius comprised 25% of isolates, E. fossor 11% and other Gram-positive rods (excluding Clostridium sp.) 18% of isolates. The Gram-negative rods comprised B. fragilis 5%, B. heparinolyticus 5%, asaccharolytic pigmented Bacteroides 3% and other Bacteroides 13%, while a so-far unnamed species of Fusobacterium (7%), and Gram-negative corroding rods (3%) were isolated. Among the facultatively anaerobic isolates, S. equi subsp. zooepidemicus accounted for 31% of isolates, followed by Pasteurella spp. 19%, Escherichia coli 17%, Actinomyces spp. 9%, Streptococcus spp. 9%. Incidental facultative isolates were Enterococcus spp. 2%, Enterobacter cloaceae 2%, Actinobacillus spp. 2% and Gram-negative corroding rods 5%. On the basis of the similarities (as determined by DNA hybridization data and/or phenotypic characteristics) of some of the bacterial species (e.g. E. fossor and B. heparinolyticus) isolated from both the normal pharyngeal tonsillar surfaces and LRT and PO diseases of horses, it is considered that the most likely source of bacteria involved in these disease processes is flora from the oral cavity.     

Osteomyelitis in dogs and cats caused by anaerobic bacteria

Localised osteomyelitis was diagnosed in 2 dogs and 2 cats. The disease was caused by fight wounds in 3 cases. Radiographic examination demonstrated a circumscribed zone of cortical bone lysis, sequestra and periosteal new bone. Each case was treated surgically by sequestrectomy and debridement. Infection was due mainly to anaerobic bacteria. The pathogenic bacteria isolated from the lesions of dogs were Actinomyces viscosus, Fusobacterium nucleatum and Bacteroides spp, and from the lesions in cats were Clostridium villosum , Peptostreptococcus anaerobius, Wolinella recta and Bacteroides gingivalis. As all the bacteria were sensitive to penicillin, each case was treated with penicillin and by irrigation of the wound. This resulted in resolution of the disease, within 4 weeks, in all cases.     

Isolation and identification of anaerobes in the veterinary diagnostic laboratory.

In a 1-year survey (1975 to 1976) of anaerobic bacteria recovered from diseased animals, anaerobes were as follows: Clostridium spp, 50%; gram-positive nonspore-forming anaerobic bacilli, 19%; gram-negative anaerobic bacilli, 19%; Actinomyces spp, 10%; and anaerobic cocci, 1%. Anaerobes were in approximately 61% of the specimens that were culturally positive for any bacteria. Approximately 25% of the specimens did not yield any bacteria (sterile specimens). The method for isolating and identifying anaerobes was based upon the use of reducible solid mediums and was specially designed for veterinary diagnostic laboratories. Emphasis was placed on the selection, collection, and transportation of specimens.     

Prevalence of Diarrhea and Enteropathogens in Racing Sled Dogs

Background: Diarrhea is highly prevalent in racing sled dogs, although the underlying causes are poorly understood.
Hypothesis: Clostridium perfringens enterotoxin (CPE) and Clostridium difficile Toxin A and B are associated with diarrhea in racing sled dogs.
Animals: One hundred and thirty-five sled dogs.
Methods: Freshly voided feces were obtained from 55 dogs before racing and from 80 dogs after 400 miles of racing. Samples were visually scored for diarrhea, mucus, blood, and melena. CPE and C. difficile Toxin A and B were detected by ELISA. Samples were cultured for C. perfringens, C. difficile, Campylobacter, Salmonella , and Escherichia coli 0157; Giardia and Cryptosporidium spp. were detected via immunofluorescence.
Results: Diarrhea occurred in 36% of dogs during racing, and hematochezia, fecal mucus or melena, or all 3 occurred in 57.5% of dogs. Salmonella was isolated from 78.2% of dogs before racing, and from 71.3% of dogs during racing. C. perfringens and C. difficile were isolated from 100 and 58.2% of dogs before racing, and from 95 and 36.3% of dogs during racing. Dogs were more likely to test positive for CPE during than before racing (18.8 versus 5.5%, P = .021); however, no enteropathogens or their respective toxins were significantly associated with hematochezia or diarrhea.
Conclusions and Clinical Importance: Sled dogs participating in long distance racing have a high prevalence of diarrhea and hematochezia that is not associated with common enteropathogens. It is possible that diarrhea and hematochezia represent the effect of prolonged exercise on the gastrointestinal tract.     

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.     

The identification and antimicrobial susceptibility of anaerobic bacteria from pneumonic cattle lungs.

One hundred and forty-four lungs obtained postmortem from cattle with pneumonia were cultured for anaerobic bacteria. Forty-five lungs yielded 73 anaerobic isolates belonging to 20 species. The number of isolations of anaerobes from acute fibrinous or suppurative bronchopneumonias (32.5%) was slightly lower than from similar chronic bronchopneumonias (36.5%). Anaerobes were not recovered from 15 lungs showing macroscopic changes not of bacterial origin, nor from 13 healthy lungs. The predominant genera isolated were Bacteroides, Peptococcus, Fusobacterium and Clostridium. The most common species were P. indolicus (15 isolates), B. asaccharolyticus (nine), F. necrophorum (six), C. perfringens (four) and B. fragilis (four). There was a significant correlation between the presence of Corynebacterium pyogenes (p less than 0.001) or Escherichia coli (p less than 0.01) and the presence of anaerobes in the lungs. The isolated anaerobic bacteria were generally susceptible to ampicillin, penicillin G, cefoxitin, cephalothin, clindamycin, chloramphenicol, erythromycin, tetracycline and metronidazole. The B. fragilis and C. perfringens isolates showed multiple antibiotic resistance, and five P. indolicus isolates were resistant to tetracycline.     

Nosocomial diarrhea associated with enterotoxigenic Clostridium perfringens infection in dogs

A retrospective analysis of the medical records of 30 consecutive cases of diarrhea occurring in dogs that were hospitalized in a teaching hospital was performed. A prospective analysis of culture results for Clostridium perfringens of dogs with diarrhea were compared with those of a control nondiarrheal group. Hospital-acquired diarrhea in dogs was found to be associated with multiple serotypes of enterotoxigenic Clostridium perfringens. Other potential etiologic agents could not be isolated. Clinical signs were variable, and included mild depression, anorexia, and soft to watery diarrhea with or without frank blood, mucus, and tenesmus. Fever was not present. There were no hematologic or serum biochemical abnormalities, nor were there any consistent virologic or parasitologic findings. Salmonella spp or Campylobacter spp were not identified by fecal culture. No risk factors could be identified. A dog that was euthanatized on the day it developed diarrhea had intestinal histologic findings suggestive of clostridial enteritis. Dogs with diarrhea had significantly higher fecal clostridial counts than did dogs without diarrhea (mean log10 counts +/- SD = 6.34 +/- 1.79 vs 4.75 +/- 2.07). Enterotoxin was found in the feces of 41% of diarrheic dogs but in only 7% of dogs without diarrhea.     

Changes in prevalence and susceptibility of obligate anaerobes in clinical veterinary practice

Of 3,133 clinical specimens obtained from domestic animals, 26% contained species of bacteria that were obligate anaerobes. Members of the genera Bacteroides, Fusobacterium, and Peptostreptococcus accounted for 77% of the isolates. On the average, 2 species of obligate anaerobes were found in each specimen, usually admixed with facultatively anaerobic bacteria. Of those specimens containing an obligate anaerobe, 20% contained one isolate that was resistant to penicillin, ampicillin, and cephalothin. These resistant isolates belonged to the genus Bacteroides, the most common being Bacteroides fragilis. Approximately one third of these penicillin-resistant isolates was resistant to tetracycline as well. All isolates were susceptible to chloramphenicol, tetracycline (excluding penicillin-resistant Bacteroides), metronidazole, and clindamycin.