Amplification of the 16S-23S r spacer region for rapid detection of Clostridium chauvoei and Clostridium septicum

Amplification of the 16S-23S rDNA spacer region by polymerase chain reaction (PCR) was used for the rapid detection of Clostridium chauvoei and C septicum. To assess its specificity, PCR was performed with total DNA from 42 strains of clostridia and three strains of other genera. PCR products specific to C chauvoei or to C septicum were generated from homologous cultures only. Clostridium chauvoer-specific or C septicum-specific amplicons were also generated from tissues of cows experimentally infected with C chauvoei or C septicum and in DNA samples from cows clinically diagnosed as having blackleg or malignant oedema. These results suggest that a species-specific PCR may be useful for the rapid and direct detection of C chauvoei and C septicum in clinical specimens.     

Demonstration of Clostridium septicum infection in a goose flock.

Clostridium septicum infection causing 5.0 to 5.2% mortality is reported for the first time in the literature from six-week-old growing geese in three flocks comprising 5,200, 5,500 and 5,900 geese, respectively. The affected birds exhibited weakness, uncoordinated movement, ataxia and, frequently, oblique position of the head and neck (torticollis) as well as signs indicative of dysequilibrium. The affected birds died within 18-24 h. Gross pathological examination revealed anaemia, hepatitis with map-like necroses of irregular outline (Fig. 1), acute enteritis, pulmonary oedema and cardiac dilatation. Light and electron-microscopic examination showed that the sinusoids of the liver were markedly dilated (Fig. 2) and filled with serous exudate and gas (Figs 2 and 3), and the hepatocytes surrounding them exhibited severe oedema (Fig. 4). Among the hepatocytes, ciliated bacteria 7-10 mu in length and 1-3 mu in width, bounded by a well-defined cell wall and often showing signs of spore formation were observed (Figs 5 and 6). By bacteriological examination the pathogen was isolated, its properties were studied, and the clinical entity of malignant oedema was experimentally reproduced by intramuscular injection of guinea-pigs and rabbits. The applied antibiotic (oxytetracycline) and furazolidone therapy proved effective.     

Tetracycline-resistance genes of Clostridium perfringens, Clostridium septicum and Clostridium sordellii isolated from cattle affected with malignant edema

The minimal inhibitory concentrations (MICs) of 10 antimicrobial agents against a total of 33 isolates of Clostridium perfringens, Clostridium septicum and Clostridium sordellii from cattle affected with malignant edema in Japan was determined. The low MIC activities of benzylpenicillin confirm the place of benzylpenicillin as the antibiotics of choice for treatment of malignant edema. Five (22%) of 23 C. septicum strains, five (71%) of seven C. perfringens strains and all strains of C. sordellii showed resistance to oxytetracycline. These oxytetracycline-resistant strains carried tetracycline-resistance genes [tetA(P), tetA408(P), tetB(P) and tetM]. The sequences of the tetracycline-resistance genes of some C. septicum strains were completely or nearly completely identical to those of strains belonging to other clostridiual species. This is the first report of resistance of C. septicum to tetracycline.     

Septic peritonitis in a Percheron mare associated with Clostridium haemolyticum

This report describes an 11‐year‐old Percheron mare that presented with signs of colic and was ultimately diagnosed with bacterial peritonitis of unknown origin. Bacterial culture of a peritoneal fluid sample isolated 2 Clostridial species, one of which was strongly suspected to be Clostridium haemolyticum. The horse was markedly hypoalbuminaemic at presentation, leading to the development of low oncotic pressure and ventral oedema. The mare was administered a low molecular weight/low molar substitution hydroxyethyl starch solution in conjunction with other therapies that resulted in marked improvement of clinical signs. The purpose of this report is to describe the clinical findings associated with equine peritonitis associated with C. haemolyticum, a rarely identified pathogen in the horse. Secondly, this report serves to describe the beneficial effects of tetrastarch administration in a clinical case with severe hypoalbuminaemia and ventral oedema.     

Diagnosis of anthrax and para-anthrax with reference to immunofluorescence technic]

504 diagnostic samples were subjected to a fluorescence serology and culturah test for black quarter and malignant oedema. In 459 cases (91.1%) the results of both testing methods wede identical. 26 first results were made by culture (14 X malignant oedema, 12 X black quarter). 19 results (14 malignant oedema, 5 X black quarter) were obtained by immuno-fluorescence only. Culture was shown to be not significantly better than fluorescence, especially in determining black quarter. In the diagnosis of black quarter and malignant oedema the combination of immunofluorescence with culture can be considered as the optimal testing method available at the present time, in that by this method there would be no need for an additional diagnostic test on the animal.     

Clostridium fallax infection in poultry

Summary A disease causing high mortality in young chickens following listlessness, inappetence and diarrhoea is described from Malawi. Haemorrhagic enteritis, splenic enlargement and congestion with petechial haemorrhages, and subcutaneous oedema were seen at post mortem.Clostridium fallax was isolated from affected birds. Coccidiosis was a possible predisposing factor.     

Anaerobic peritonitis caused by Clostridium septicum as a complication of routine castration in a 2‐year‐old Warmblood horse

This case report describes an unusual case of anaerobic peritonitis in a 2‐year‐old horse following castration. The horse was evaluated 2 weeks following castration for signs of acute, severe abdominal pain and swelling surrounding a previous castration site. Physical examination revealed marked scrotal and ventral abdominal oedema that was cool and crepitant upon palpation. Ultrasonographic evaluation was unrewarding because gas shadowing distributed throughout the subcutis prevented imaging of the abdominal cavity. Ventral midline celiotomy revealed a copious amount of malodorous, serosangious, cloudy peritoneal fluid that was submitted for culture. Abdominal exploration revealed the gastrointestinal tract to be in its anatomically correct position. There was diffuse petechiation of the small intestine and large intestine, oedema and crepitant swelling surrounding the left inguinal ring and body wall. The abdomen was lavaged with 10 l of sterile saline prior to closure of the celiotomy and the left castration incision was opened digitally, releasing a large volume of serosanguinous fluid and gas that flowed freely from the incision site and deeper inguinal tissues. The horse was placed in the recovery box where it suffered cardiac arrest. Culture of the peritoneal fluid revealed heavy growth of Clostridium septicum. This case of anaerobic peritonitis represents an unusual complication following castration not previously reported in the horse.     

The effect of enterotoxin of Clostridium welchii (perfringens) type A on fowls.

Lethal doses of enterotoxin of Clostridium welchii (perfringens) type A injected intravenously into young fowls caused immediate lassitude, with partial recovery, followed by death seven to 35 h after inoculation. Lesions found were ascites, hydropericardium, oedema of the muscles, hepatic congestion, urate deposits on the peritoneum and the pericardium, and intestinal hyperaemia. Sublethal doses produced no clinical signs or lesion. The LD50 of this enterotoxin was 74-84 mug/kg of body weight.     

A case of feline primary inflammatory mammary carcinoma: clinicopathological and immunohistochemical findings

The clinicopathological and immunohistochemical findings of a primary feline mammary tumour with features similar to human and canine primary inflammatory carcinoma are described for the first time. The cat presented to the clinic for the rapid onset of oedema, severe erythema, local pain and warmth of the inguinal region, with a pustular-to-nodular cutaneous lesion in association with an ill-defined underlying mass. An epithelial malignant tumour was diagnosed by cytological investigation. Necropsy revealed a thickening of the skin with oedema of the subcutis in the right inguinal area, and regional and distant metastases. Histology showed an unencapsulated tubulopapillary proliferation of malignant epithelial cells, with a massive embolisation in the dermal lymphatics and a mild inflammatory infiltrate. Through immunohistochemistry, the tumour was found to be oestrogen (ER)-alpha-, androgen (AR)- and progesterone (PR)-negative; neoplastic cells were ER-alpha, AR-negative and focally PR-positive. An irregular, mild and focal HER-2 immunoreactivity was present (score +1, non-HER-2 overexpressing). The neoplastic cells were cyclo-oxygenase-2 and vascular endothelial growth factor positive.     

Secondary malignant lymphoedema after mastectomy in two dogs

This case report describes the diagnosis of secondary malignant lymphoedema in two dogs that had undergone a mastectomy. A remarkable severe oedematous lesion associated with lameness in the right hindlimb was observed in both cases. Diagnostic imaging examinations, including direct pedal lymphangiography (case 1) and lymphoscintigraphy (case 2), showed obstruction of lymph flow in the lymphatics of the right hindlimbs. Although the recommended medical management and physiotherapy had been applied to resolve the problems, oedema did not improve in the damaged region in both cases. Results of histopathological examinations suggested that the cause of the obstructed lymph flow was neoplastic cells in the lymphatics of the right hindlimb in both dogs.     

Generalised peripheral oedema associated with amlodipine therapy in two dogs

This report details two cases of adverse drug reactions to amlodipine. The first case presented with diffuse peripheral oedema and a history of amlodipine therapy. Haematology, clinical chemistry, endocrine testing, thoracic, abdominal and cardiac imaging revealed no cause for oedema. Amlodipine therapy was discontinued and oedema diminished markedly within 72 hours. The second case presented for bilateral retinal detachments secondary to systemic hypertension. Haematology, clinical chemistry, thoracic and abdominal imaging were unremarkable and amlodipine therapy was begun. Within 72 hours, diffuse peripheral oedema developed that was unresponsive to therapy and the dog was euthanised. Veterinarians should be aware of the potential serious adverse events associated with commonly used drugs; severe, diffuse oedema is a possible adverse drug event in dogs treated with amlodipine .     

Tyzzer's disease in a Eurasian otter (Lutra lutra) in Scotland

A Eurasian otter (Lutra lutra) cub found in weak condition on the Isle of Harris, Scotland, developed bilateral corneal oedema 16 days after being admitted to a rehabilitation centre. It died unexpectedly on day 26. On postmortem examination, there was excess clear fluid in the body cavities and the liver was swollen with numerous pale focal lesions and petechial haemorrhages throughout. Histopathological examination revealed bundles of bacilli morphologically typical of Clostridium piliforme within hepatocytes. Comparative analysis of the nucleotide base sequence of a 16S rdna fragment amplified from the infected liver tissue revealed that it was identical to a C piliforme 16S rdna sequence. The possibility of concurrent infection with canine adenovirus type 1 was considered but none of the characteristic histopathological lesions was observed and examination of the liver by transmission electron microscopy was negative for virus particles. This appears to be the first record of Tyzzer's disease in an otter and the first in a wild animal in Britain.     

Pathogenesis of brain damage produced in sheep by Clostridium perfringens type D epsilon toxin: a review

Microvascular endothelial damage by the epsilon toxin of Clostridium perfringens type D appears to be the fundamental cause of cerebral parenchymal injury and lesions occur in a seemingly dose- and time-dependent manner. Large doses of circulating toxin produce a severe, generalised, vasogenic cerebral oedema and an acute or peracute clinical course to death. With lower doses of toxin, or in partially immune sheep, focal necrosis, often bilaterally symmetrical, occurs in certain selectively vulnerable brain regions, which appear to become fewer as the toxin dose is reduced. These cases follow a more protracted clinical course, but death is the usual outcome. The precise pathogenesis of the focal brain damage found in subacutely intoxicated sheep is unresolved, but several possible mechanisms are discussed.     

Alternative method to facilitate resolution of paraphimosis after penile trauma in the horse

Inflammation associated with traumatic injury to the penis is characterised by pain, heat and oedema. Ensuing penile swelling interferes with retraction of the penis into the prepuce, resulting in paraphimosis. This report describes the use of a 5 l capacity pressure infusion bag to reduce penile oedema in three horses. This technique provided complete resolution of oedema and associated paraphimosis in approximately 1 h. The use of the bag, along with an abdominoinguinal bandage, a plastic funnel suspensory device, or a penile repulsion device (Probang) to avoid recurrence, offers a novel treatment that may be performed with the horse sedated.     

Leukoencephalomalacia in a horse induced by fumonisin B1 isolated from Fusarium moniliforme

Each of two horses was dosed by stomach tube with culture material on maize of Fusarium moniliforme MRC 826. One horse developed severe hepatosis and mild oedema of the brain after 6 doses of 2.5 g of culture material/kg body mass/day in 7 days. The second horse, in a similar experiment but at a dosage rate of 1.25 g/kg/day, developed mild hepatosis and moderate oedema of the brain. In both animals the brain oedema was particularly noticeable in the medulla oblongata. The mycotoxin fumonisin B1 was extracted and purified from the culture material of F. moniliforme MRC, 826 which contained approximately 1 g/kg of this compound. A horse was injected intravenously 7 times from Day 0-Day 9 with 0.125 mg of fumonisin B1/kg body mass/day. Clinical signs of neurotoxicosis, which appeared on Day 8, included nervousness followed by apathy, a wide-based stance, trembling, ataxia, reluctance to move, paresis of the lower lip and tongue, and an inability to eat or drink. Euthanasia was performed on the horse on Day 10 while the animal was in a tetanic convulsion. The principal lesions were severe oedema of the brain and early, bilaterally symmetrical, focal necrosis in the medulla oblongata. This report provides experimental evidence that fumonisin B1, produced by F. moniliforme, causes equine leukoencephalomalacia.     

Cutaneous T-cell-rich B-cell lymphoma in a horse

Cutaneous malignant lymphomas are rare in horses and comprise predominantly T-cell-rich B-cell lymphomas. They are characterized by multiple tumour nodules affecting predominantly female horses with a survival rate of months to years. At the final stage, metastases to regional lymph nodes occur, whereas widespread organ involvement is rarely reported. In this case report, a cutaneous T-cell-rich B-cell lymphoma in a 7-year-old standardbred gelding with metastases is described. Clinically, multiple cutaneous and subcutaneous nodules, enlarged superficial lymph nodes, rapid weight loss, and ventral oedema were observed. In addition to the clinical findings, necropsy revealed tumour infiltration in multiple body lymph nodes, a solitary pleural mass, and few pulmonary and intestinal tumour nodules. Microscopically, all neoplasms were composed of a densely packed cell population consisting of large lymphoblastic cells expressing CD79a, and numerous small, round, CD3-positive T lymphocytes. With respect to these findings the diagnosis of a cutaneous T-cell-rich B-cell lymphoma with metastases was made.     

Proteases of Clostridium botulinum. V. Studies on the serological relationship between proteases from Clostridium botulinum and other spore-forming bacteria

By the use of the electrophoretic casein precipitating inhibition test (CPI-test) the serological relationship between proteolytic enzymes produced by different species within the genera Clostridium and Bacillus has been tested. The proteases produced by Clostridium botulinum types A, B, C, D and F cross-reacted with each other. Clostridium botulinum strain 84 was inhibited by antiproteases produced against Clostridium sporogenes, Clostridium botulinum types C and F (protease F I and F II), but not by antiproteases against Clostridium botulinum types B and F (protease II), Clostridium bifermentans and Clostridium perfringens. The protease of the newly described Clostridium botulinum strain 89 (type G) was inhibited by Clostridium sporogenes antiprotease, but not by any of the other antiproteases. It is not possible to differentiate between Clostridium botulinum, Clostridium sporogenes and Clostridium perfringens by use of serological differentiation of their proteolytic enzymes. The protease of Clostridium bifermentans is not serologically related to any of the species tested in this investigation. Proteases produced by different Bacilli were not inhibited by antiproteases from Clostridium botulinum types B, C and F, Clostridium sporogenes, Clostridium bifermentans, and the two strains of Clostridium perfringens tested. This investigation indicates a serological relationship between proteases from different Clostridium species, but not a serological relationship between proteases produced by the Clostridium species and Bacillus species tested.     

A case of nontraumatic gas gangrene in a common marmoset (Callithrix jacchus)

The common marmoset is widely used in neuroscience and regenerative medicine research. However, information concerning common marmoset disorders, particularly infectious diseases, is scarce. Here, we report a case of a female common marmoset that died suddenly due to gas gangrene. The animal presented with gaseous abdominal distention at postmortem, and Clostridium perfringens type A was isolated from several tissues. Vacuoles, a Gram-positive bacteremia and intravascular hemolysis were observed microscopically in the muscles, liver and lungs. On the basis of these findings, we diagnosed nontraumatic gas gangrene caused by Clostridium perfringens type A in this common marmoset.     

Severe chemosis caused by nephrotic syndrome in a goat: a case report

A 3-year old female goat with nephrotic syndrome was presented because of severe bilateral chemosis. The diagnosis was based on clinical findings, decreased packed cell volume, increased serum urea and creatinine, decreased serum protein and protein loss via the urinary tract. At post mortem examination, a chronic glomerulonephritis and interstitial nephritis were found. In addition to severe conjunctival oedema, ascites and subcutaneous oedema in the region of the larynx were noted. Immunohistochemistry for the detection of chlamydial antigen in the conjunctiva was negative. This is the first report of severe chemosis caused by nephrotic syndrome in a goat.     

Malignant melanoma of the pelvis as a possible cause of pelvic limb lameness in two gray mares

Two grey mares, a 12-year-old Arabian and a 13-year-old Lippizaner cross, were referred for chronic pelvic limb lameness and abduction of the limb during the non supporting phase of the stride. The clinical examination revealed melanomas under the tail and on the perineum, and oedema of the proximal lame limb. A mass in the pelvic area was detected on rectal palpation. Laparoscopy was performed and the provisional diagnosis of malignant melanoma was made. Histological examination of the biopsy taken under laparoscopic control from the mass in the pelvis confirmed the diagnosis of malignant melanoma. In the postmortem examination a large black mass in the pelvis and metastases in several organs were diagnosed. The process under the tail showed microscopically the characteristics of dermal melanomatosis. The metastases showed the appearance of a malignant melanoma. The lameness and abduction during the non supporting phase of the stride were possibly caused by a combination of pain, a mechanical limitation, and paresis/paralysis. When an older grey horse with melanomas on the predilection sites is submitted with pelvic limb lameness and abduction of the limb during the non supporting phase of the stride, a malignant melanoma in the pelvis is one of the differential diagnoses and a rectal examination should be performed. Diagnostic laparoscopy in the standing horse can be useful to confirm the provisional diagnosis.