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Relationship between resistance of salmonids to furunculosis and recovery of Aeromonas salmonicida from external mucus.

Fish were sampled at the Ed Weed State Fish Hatchery (South Hero, Vermont, USA) in September 1992. Aeromonas salmonicida was common, with concentrations as high as 10(5) to 10(7) colony-forming units per gram of mucus, and readily recovered from most mucus samples obtained from furunculosis-sensitive populations of brook trout (Salvelinus fontinalis), lake trout (Salvelinus namaycush), and Atlantic salmon (Salmo salar). The pathogen was the predominant microorganism and accounted for greater than 85% of the total number of bacteria isolated from the mucus of these fish. By comparison, A. salmonicida was recovered only from two rainbow trout (Oncorhynchus mykiss), and bacterial frequencies did not exceed 10(3) colony-forming units per gram of mucus. The pathogen was not recovered from the mucus of steelhead (O. mykiss) or Rome brown trout (Salmo trutta) selectively bred for resistance to furunculosis, even though there was widespread contagion throughout the hatchery and fish were cultured on a common, unprotected water supply.

Aeromonas↗

Furunculosis control using a paraimmunization stimulant (Baypamun) in rainbow trout.

The effect of Baypamun, a non-specific immune system stimulant, on the treatment and control of furunculosis was studied in experimentally infected rainbow trout. A statistically significant association between the Baypamun treatment and a reduction in cumulative incidence of the clinical symptoms of Furunculosis and the infection level, despite the presence of the bacteria in blood and organs was observed. A reduction in the mortality rate in Baypamun-treated fish was also observed but this was not statistically significant. The results suggest that the treatment of fish diseases with non-specific immune stimulants could be, in the future, a good defense strategy leading to a reduction in disease symptoms and a greater control of the asymptomatic carriers after an outbreak.

Adjuvants, Immunologic↗

Dietary intake of immunostimulants by rainbow trout affects non-specific immunity and protection against furunculosis.

Immunostimulant preparations Macrogard, Candida utilis, Saccharomyces cerevisiae, Evetsel, Chitosan, or FinnStim were mixed into semipurified diets and fed to groups of rainbow trout for 1 week. Fish were bled by non-lethal caudal puncture and blood samples assayed for changes in non-specific cellular immunity and humoral protein levels. In the immunostimulated fish, hematocrit levels and lymphocyte counts remained relatively stable; however, elevations were observed in oxidative radical release, myeloperoxidase activity, phagocytic indexes, and potential killing activities of phagocytic cells including neutrophils. Total plasma protein and total immunoglobulin levels were elevated by the dietary immunostimulants. A challenge with the virulent pathogen that causes furunculosis, Aeromonas salmonicida, showed that the immunostimulated groups of fish were more resistant to the disease, confirming the potential use of these substances in fish culture for the prevention of disease.

Adjuvants, Immunologic↗

Differential susceptibility of salmonid fishes to furunculosis correlates with differential serum enhancement of Aeromonas salmonicida extracellular protease activity.

Rainbow trout, Atlantic salmon and brown trout, respectively, exhibit an increasing order of susceptibility to furunculosis. We investigated if this may be due to a differential sensitivity to exotoxins or ability to inhibit the exoprotease which is required by the bacterium to digest protein for growth. Differences were slight in susceptibility to exotoxins and serum inhibition of exoprotease at high exotoxin:serum ratios. However, at low ratios brown trout serum markedly enhanced protease activity, salmon serum had little effect and rainbow trout serum maintained 50% inhibition. This implies that during the early stages of infection the potential for growth of the bacterium would be highest in brown trout and lowest in rainbow trout. Determinations of growth rates in fresh serum support this hypothesis. The enhancement of exoprotease activity by serum of susceptible species may represent an important virulence mechanism.

Aeromonas↗

Matrix metalloproteinase mRNA expression in canine anal furunculosis lesions.

Although the aetiology of anal furunculosis (AF) in dogs is poorly understood, there is evidence for an underlying immune dysfunction. This is illustrated by the presence of a T helper type 1 cytokine mRNA profile in AF lesions and the clinical response to ciclosporin therapy. Expression of MMPs 2, 9 and 13 were evaluated in AF lesional biopsies by real-time quantitative RT-PCR. There was significantly increased expression of both MMP-9 and MMP-13 mRNA in AF biopsies compared to controls (p<0.001) but no significant difference in MMP-2 mRNA expression. Since MMP-9 and MMP-13 are primarily produced by macrophages, these data suggest that ulceration could be the result of aberrant activation of this cell type in the tissues. It is feasible that such pathological macrophage activity occurs in response to interferon-gamma secreted by T helper type 1 cells. This could explain why the lesions resolve following treatment with the immunosuppressive drug ciclosporin.

Animals↗

Aspects of the humoral immune response to Staphylococcus intermedius in dogs with superficial pyoderma, deep pyoderma and anal furunculosis.

Bacterial infection (pyoderma) of the canine skin is largely caused by Staphylococcus intermedius and may be a superficial or deep infection. Pyoderma may be a primary, idiopathic disease or secondary to a range of other dermatological disorders. In this study, the serum concentrations of IgG, IgA, antistaphylococcal IgG and antistaphylococcal IgA were measured by ELISA in normal dogs (n = 22), dogs with idiopathic deep pyoderma (n = 22), atopic dermatitis and superficial pyoderma (n = 24), atopic dermatitis without pyoderma (n = 25), flea bite dermatitis with superficial pyoderma (n = 8), pustular demodicosis (n = 8) and German shepherd dogs with anal furunculosis (n = 28). The serum IgG was significantly increased in dogs with atopy and superficial pyoderma (p < 0.001), and lower than normal in dogs with idiopathic deep pyoderma (p < 0.015). The concentration of serum IgA was significantly lower than normal in dogs with atopy uncomplicated by pyoderma (p < 0.015). The concentration of antistaphylococcal IgG in all clinical sera was significantly elevated (p < 0.001) when compared to normal dogs but concentrations of antistaphylococcal IgA were no greater than in normal dogs. Western blotting analysis for determination of the specificity of serum IgG antistaphylococcal antibody revealed that there were nine major epitopes. Discriminant analysis demonstrated that particular combinations of these epitopes were recognised more frequently by sera from dogs in different clinical groups.

Animals↗

Synergistic effects of dietary iron and omega-3 fatty acid levels on survival of farmed Atlantic salmon, Salmo salar L., during natural outbreaks of furunculosis and cold water vibriosis.

The present study demonstrates that farmed Atlantic salmon, Salmo salar, health is positively and significantly affected by synergistic effects between very long-chain polyunsaturated fatty acids of the n-3 family eicosapentaenoic acid/docosahexaenoic acid (EPA/DHA) and iron, where positive effects of high dietary levels of EPA/DHA are enhanced when combined with low levels of iron. Based on cumulative mortalities in the different experimental groups, relative percentage of survival (RPS) for the high EPA/DHA-low iron group was 70% during an outbreak of furunculosis and 96% during an outbreak of cold water vibriosis compared with the controls. A non-additive effect between EPA/DHA and iron was confirmed by statistical analyses that revealed a significant effect of EPA/DHA alone and an interaction of iron with EPA/DHA. Liver cell cultures treated with EPA/DHA revealed that the synergistic effect could be related to an EPA/DHA dependent regulation of mRNA for proteins important for transport (transferrin) and storage (ferritin) of iron in the salmon. In keeping with this finding, the transcriptional down-regulation of iron metabolism in vitro was reflected in decreased in vivo iron stores with increasing levels of dietary EPA/DHA. Hence, to avoid overloading of the iron transport/storage-systems resulting in increased susceptibility to bacterial infections, high levels of dietary EPA/DHA should be accompanied by low levels of dietary iron.

Animals↗

The role of surgery in the management of canine anal furunculosis. A review of the literature and a retrospective evaluation of treatment by surgical resection in 51 dogs.

AIMS: To retrospectively evaluate the outcome of surgical management of anal furunculosis (AF) in 51 dogs. To compare the outcome of surgery with those of current medical protocols by way of a review of the literature. METHODS: Dogs referred for treatment of AF (n=51) were treated by en bloc surgical resection of diseased tissue and primary wound closure. This technique was combined with bilateral anal sacculectomy in all cases in which the anal sacs had not been previously removed. Immunomodulatory therapies were not used, with the exception of post-operative metronidazole antibiosis for 7-10 days. Follow-up was performed by the author using a telephone questionnaire. RESULTS: Forty-eight dogs were eligible for post-operative follow-up 1.5 to 36 (mean 17.4, median 18) months after surgery. Lesion recurrence, faecal incontinence and stricture formation occurred in 2%, 4% and 13% of dogs, respectively. The percentage of dogs considered by their owners to have an acceptable level of faecal continence and an improved quality of life was 94%. CONCLUSIONS AND CLINICAL RELEVANCE: The aetiopathogenesis of canine AF remains unclear. Whilst recent advances in medical management by the use of various immunomodulatory medications (such as cyclosporine) hold promise, this approach has yet to be refined with respect to affordability, long-term efficacy and morbidity. With meticulous surgical dissection and reconstruction techniques (anoplasty), excellent success rates can be achieved following a single surgical procedure with minimal complications. Surgery remains a viable treatment option, alone or in combination with immunomodulatory medications, until a more thorough understanding of this debilitating disease is achieved.

Anal Sacs↗

Strategies for the management of recurrent furunculosis.

Furuncles are common infections of hair follicles. Staphylococcus aureus is frequently the causative agent, though other bacteria may also be pathogenic, especially for furuncles in the vulvovaginal area, the perirectal area, and the buttocks. A simple furuncle can be treated by incision and drainage. Systemic antibiotics are indicated in only special circumstances, and Gram stain is helpful in choosing an appropriate one. Some patients are plagued by recurrent furuncles because of follicular abnormalities, climatic conditions, colonization by pathogenic strains, reinfection, debility, or immunodeficiency. I present various strategies for treatment and prevention of recurrent furunculosis.

Furunculosis↗

Juvenile pityriasis rubra pilaris associated with hypogammaglobulinaemia and furunculosis.

We report a case of follicular keratosis with inflammatory changes, consistent with a diagnosis of atypical juvenile pityriasis rubra pilaris. An unusual feature was the occurrence of severe Staphylococcus aureus folliculitis and furunculosis, a phenomenon rarely encountered in pityriasis rubra pilaris and the other follicular keratoses. Standard antibiotic and antiseptic treatment for chronic S. aureus infection was ineffective. The patient was subsequently found to have hypogammaglobulinaemia, and treatment with human polyvalent immunoglobulin infusions was successful in eradicating the sepsis. It is therefore probable that the hypogammaglobulinaemia played a pathogenic role in the development of cutaneous sepsis.

Adult↗

Preoperative immunosuppressive therapy and surgery as a treatment for anal furunculosis.

OBJECTIVES: To evaluate the efficacy of combining preoperative immunosuppressive therapy with surgical excision for treatment of anal furunculosis (AF) in dogs. STUDY DESIGN: Retrospective study. ANIMALS: Dogs (n=25) with stages 1-4 AF. METHODS: Preoperative immunosuppressive therapy was either cyclosporine A (CyA) alone or in combination with ketoconazole (Group 1; n=18), or azathioprine combined with prednisolone (Group 2; n=7). Surgical excision of residual draining tracts, cryptectomy, and anal sacculectomy were performed. Only dogs with postoperative clinical follow-up exceeding 9 months were retained for the study. RESULTS: Both immunosuppressive protocols were effective in reducing progression of AF. Subsequent draining tracts excision, cryptectomy, and anal sacculectomy were bilateral (12 dogs) or unilateral (13 dogs of which 4 had bilateral anal sacculectomy). Postoperative recovery was uneventful, except for 2 dogs that had wound breakdown. Recurrence was not observed in any of the dogs that had bilateral surgical excision and or in 9 dogs that had unilateral excision. CONCLUSION: Preoperative immunosuppressive therapy, combined with bilateral surgical resection of affected tissue consistently, resulted in resolution of AF. Four dogs that had recurrence had unilateral excision despite initial bilateral involvement, suggesting that all diseased tissue should be excised. CLINICAL RELEVANCE: These preliminary results suggest that immunosuppressive therapy before surgical excision for AF yields minimizes recurrence in dogs.

Anal Sacs↗

Cyclosporin as the sole treatment for anal furunculosis: preliminary results.

Cyclosporin was used, as the sole agent, to treat six cases of anal furunculosis, allowing the clinical response to the drug to be evaluated. The initial dose was 7.5 mg/kg twice a day. Following one week of treatment, whole blood cyclosporin trough levels were measured and the dose was adjusted to obtain a stable trough level of 400 to 600 ng/ml. In all dogs, the lesions were found to follow a similar pattern of resolution throughout the course of treatment. The presenting signs disappeared within one week of treatment, with a reduction in lesion size of 50 to 90 per cent. Lesions then gradually healed over the remainder of the treatment, which lasted from 10 to 20 weeks. One case required cryosurgical treatment to resolve a 2 mm diameter persistent ulcerated lesion following 18 weeks of treatment. The length of follow-up was four to 14 months (mean 7.7 months). One case developed recurrence of the lesions eight weeks post-treatment.

Animals↗

Efficacy of combined cyclosporine A and ketoconazole treatment of anal furunculosis.

Cyclosporine A and ketoconazole were used as a combined therapy to treat 19 dogs with anal furunculosis. Complete resolution of all lesions was achieved in three to 10 weeks, but recurrences occurred in seven of the 19 dogs (36.8 per cent), with remission periods extending from one to six months for these dogs. Adverse effects of treatment included excessive hair loss, intermittent lethargy, vomiting and decreased appetite in some dogs, but none of the signs were considered serious. The results of treatment are comparable with, if not better than, the surgical alternatives. There is an approximate 70 per cent cost saving over the use of cyclosporine alone.

Animals↗

An aromatic-dependent mutant of the fish pathogen Aeromonas salmonicida is attenuated in fish and is effective as a live vaccine against the salmonid disease furunculosis.

Aeromonas salmonicida is the etiological agent of furunculosis in salmonid fish. The disease is responsible for severe economic losses in intensively cultured salmon and trout. Bacterin vaccines provide inadequate protection against infection. We have constructed an aromatic-dependent mutant of A. salmonicida in order to investigate the possibility of an effective live-attenuated vaccine. The aroA gene of A. salmonicida was cloned in Escherichia coli, and the nucleotide sequence was determined. The codon usage pattern of aroA was found to be quite distinct from that of the vapA gene coding for the surface array protein layer (A layer). The aroA gene was inactivated by inserting a fragment expressing kanamycin resistance within the coding sequence. The aroA::Kar mutation was introduced into the chromosome of virulent A. salmonicida 644Rb and 640V2 by allele replacement by using a suicide plasmid delivery system. The aroA mutation did not revert at a detectable frequency (< 10(-11). The mutation resulted in attenuation when bacteria were injected intramuscularly into Atlantic salmon (Salmo salar L.). Introduction of the wild-type aroA gene into the A. salmonicida mutants on a broad-host-range plasmid restored virulence. A. salmonicida mutant 644Rb aroA::Kar persisted in the kidney of brown trout (Salmo trutta L.) for 12 days at 10 degrees C. Vaccination of brown trout with 10(7) CFU of A. salmonicida 644Rb aroA by intraperitoneal injection resulted in a 253-fold increase in the 50% lethal dose (LD50) compared with unvaccinated controls challenged with a virulent clinical isolate 9 weeks later. A second vaccination after 6 weeks increased the LD50 by a further 16-fold.

4-Aminobenzoic Acid↗

A live (delta aroA) Aeromonas salmonicida vaccine for furunculosis preferentially stimulates T-cell responses relative to B-cell responses in rainbow trout (Oncorhynchus mykiss).

We have previously described (L. M. Vaughan, P. R. Smith, and T. J. Foster, Infect. Immun. 61:2172-2181, 1993) the construction of a kanamycin-resistant aromatic-dependent mutant of Aeromonas salmonicida, the causative agent of furunculosis, and its use as a live vaccine. Here we describe the construction of an unmarked aroA deletion mutant and examine the nature and magnitude of immune responses in rainbow trout (Oncorhynchus mykiss) to this vaccine strain. Good proliferative and antibody responses were elicited by using a range of vaccine doses from 2 x 10(6) to 2 x 10(9) live bacteria per fish, and a clear vaccine dose effect was apparent. A significant positive effect of using live bacteria to prime for lymphocyte proliferation and antibody production was apparent. However, the problem of directly comparing the vaccine doses of live and killed bacterial preparations is discussed, since some replication of live bacteria in vivo is expected. Most importantly, the live vaccine preferentially stimulated enhanced T-cell responsiveness, as evidenced by significantly greater increases in T-cell proliferation than in B-cell proliferation, compared with responses by the respective cell populations from fish given a killed vaccine. The manner in which live vaccines elicit strong cell-mediated immune responses and the relevance to fish vaccine design are discussed.

Aeromonas↗

Furunculosis due to Mycobacterium mageritense associated with footbaths at a nail salon.

We report two cases of lower-extremity furunculosis caused by Mycobacterium mageritense. Both patients were patrons of the same nail salon, where they received footbaths prior to pedicures. M. mageritense bacteria isolated from two whirlpool footbaths were determined to be closely related to the patient isolates by pulsed-field gel electrophoresis.

Adult↗

Evaluation of once daily treatment with cyclosporine for anal furunculosis in dogs.

Twenty-four dogs with anal furunculosis were treated with cyclosporine once daily for 13 weeks at dosages of 1.5, 3.0, 5.0 or 7.5 mg/kg, and re-examined after six and 12 months. After 13 weeks the disease in six of the dogs was in remission, 11 were controlled or improved and seven had failed to respond. The response of the dogs given the highest dose was significantly better than the response of the other groups taken together (P < 0.014), and better than the responses of the groups given 1.5 mg/kg and 5 mg/kg (P < 0.05). The dogs improved clinically during the treatment, most rapidly during the first five weeks. Of the six dogs that were in remission after 13 weeks, three relapsed after one, two and six months. The 11 dogs that were improved or controlled after 13 weeks were either left untreated or were continued on cyclosporine medication for one to three months at a dosage of 1.5 to 7.5 mg/kg; the disease went into remission in four cases and remained controlled in the other seven, but four of the 11 cases relapsed during the 12 months following the treatment. The side effects observed included increased coat turnover and transient vomiting.

Adjuvants, Immunologic↗