Dysgonic strain of Microsporum canis pseudomycetoma in a Domestic Long-hair cat

A 4-year-old Domestic Long-hair cat was presented with two large non-painful, ulcerated and suppurative lesions over the flanks. Histopathology and cytology were consistent with fungal pyogranulomatous inflammation. Culture of tissue yielded a dysgonic strain of Microsporum canis. The cat was treated successfully by staged en bloc resections of the lesions, followed by oral ketoconazole, then oral terbinafine. This is the first reported case of dermatophytic pseudomycetoma in a Domestic Long-hair cat in Australia.     

Dermal mass aspirate from a Persian cat

A 1-year-old spayed female Persian cat with alopecia and weight loss had numerous variably ulcerated dermal nodules. Cytologic examination of an aspirate of one of the nodules revealed pyogranulomatous inflammation along with septate hyphae and basophilic round bodies, 0.5–1.0 μm in diameter, surrounded by a thin clear halo (arthrospores). The cytologic diagnosis was dermatophytic pseudomycetoma. Histologically, there were dermal granulomas containing poorly staining, septate hyphae with bulbous spores embedded within abundant amorphous eosinophilic material (Splendore-Hoeppli reaction), and the histologic diagnosis was pseudomycetoma-associated chronic multifocal severe granulomatous dermatitis with lymphocytic perifolliculitis and furunculosis. Microsporum canis was cultured from the lesion. Pseudomycetomas are distinguished from fungal mycetomas, or eumycotic mycetomas, by the findings of multiple lesions, lack of a history of skin trauma, an association with dermatophytes, most commonly Microsporum canis , and, histologically, lack of true cement material and a more abundant Splendore-Hoeppli reaction in pseudomycetomas. Additionally, pseudomycetomas differ from dermatophytosis, in which lesions are restricted to epidermal structures. Persian cats have a high incidence of pseudomycetoma formation, suggesting a heritable predisposition. The prognosis is fair with systemic antifungal therapy. When examining cytologic specimens from Persian cats with single or multiple dermal nodules, especially if pyogranulomatous inflammation is present, a diagnosis of pseudomycetoma should be suspected and is warranted if arthrospores and refractile septate hyphae are present.     

Imaging diagnosis--sublumbar pseudomycetoma in a Persian cat.

A 6-year-old Persian cat was examined for constipation, anorexia, and vomiting that was subsequently found to be due to a pseudomycetoma originating from the descending colon and sublumbar region, and causing mechanical obstruction of the colon and rectum. Multiple discrete hyperechoic foci likely representing fungal grains within the lesion gave the mass a coarse echotexture on ultrasound and was supportive of the diagnosis and computed tomography allowed delineation the extent of the mass. A pseudomycetoma is a granulomatous/pyogranulomatous reaction that surrounds dermatophytic fungal hyphae. Definitive diagnosis of a dermatophytic pseudomycetoma requires identification of the etiologic agent by cultivation or immunohistochemical staining. A pseudomycetoma should be included in the differential diagnosis for an abdominal mass in a Persian cat, especially is accompanied by the sonographic findings noted above.     

IMAGING DIAGNOSIS—SUBLUMBAR PSEUDOMYCETOMA IN A PERSIAN CAT

A 6-year-old Persian cat was examined for constipation, anorexia, and vomiting that was subsequently found to be due to a pseudomycetoma originating from the descending colon and sublumbar region, and causing mechanical obstruction of the colon and rectum. Multiple discrete hyperechoic foci likely representing fungal grains within the lesion gave the mass a coarse echotexture on ultrasound and was supportive of the diagnosis and computed tomography allowed delineation the extent of the mass. A pseudomycetoma is a granulomatous/pyogranulomatous reaction that surrounds dermatophytic fungal hyphae. Definitive diagnosis of a dermatophytic pseudomycetoma requires identification of the etiologic agent by cultivation or immunohistochemical staining. A pseudomycetoma should be included in the differential diagnosis for an abdominal mass in a Persian cat, especially is accompanied by the sonographic findings noted above.     

Dermatophytic pseudomycetomas in four cats

The aim of this retrospective study was to describe the clinical characteristics and treatment of four cats with dermatophytic pseudomycetoma. Four Persian cats, one female and three males, with age ranging from 1.4 to 5 years, were diagnosed with dermatophytic pseudomycetoma by histological examination and fungal culture. Wood's lamp examination revealed positive fluorescence of hairs in all four cats. Characteristic skin lesions consisted of multifocal, raised, firm and nodular to dome-shaped lesions varying in size from 1 to 8 cm in diameter, with ulcers or fistulas in some of the lesions. One cat was treated and cured with 3 months of oral itraconazole; lesions completely regressed, and at the time of writing there has been no recurrence. One cat was treated with surgical excision alone, and recurrence of lesions occurred after a disease-free interval of 15 months. Two cats were treated with surgical excision and systemic itraconazole therapy. Itraconazole therapy was started 1-2 months before surgery and continued for 3 months after surgery. Surgical margins were wide in both cats, and underlying adipose tissue and/or deeper fascia was removed. One cat relapsed, but had a disease-free interval of 18 months. The other cat has been disease free for 32 months. This case series suggests that aggressive, wide surgical excision and concurrent oral itraconazole are highly beneficial in treating dermatophytic pseudomycetoma in cats.     

A rapid agglutination assay for canine brucellosis using antigen coated beads

Brucella canis is the causative agent of canine brucellosis and facultative intracellular pathogen. The diagnosis of canine brucellosis is based on bacteriological examination and serological methods including agglutination and gel diffusion tests. In this study, crude antigens were extracted from B. canis using hot saline, coated on to latex beads and their usefulness in the serological diagnosis of canine brucellosis was examined. Mixing the antigen coated latex beads with the sera of dogs infected with B. canis produced clear agglutination, but this was not so for B. canis free dog sera. N-terminal amino acid sequence analysis of the crude hot saline extracts, showed that they contained copper-zinc superoxide dismutase, ribose ABC transporter and hypothetical protein of Brucella as antigens. A serological survey of canine serum samples conducted by means of an agglutination test using the antigen coated latex beads, showed that this method was more specific than the tube agglutination test using whole bacterial cell antigens. Although these results suggest that our method in which crude hot saline extracted antigens are coated on to latex beads would be useful in the serological diagnosis of canine brucellosis, we need further investigation using more serum samples to confirm the usefulness of our method.     

Molecular detection and characterization of Ehrlichia canis from dogs in Turkey

Seroprevalence of Ehrlichia canis antibodies among dogs in Turkey were previously reported, however, the ehrlichial organism has never been characterized in this region. The current study examined dogs from Ankara with febrile illness for E. canis infection with E. canis-specific PCR. Three of the 12 blood specimens from dogs showing clinical signs compatible with canine ehrlichiosis were found to be positive by PCR using E. canis-specific primers. E. canis detected in one of the blood specimens was designated as Kutahya strain. The representative E. canis strain was characterized by 16S rRNA gene sequencing and Western blot analysis of the plasma sample from the dog infected with E. canis. The 16S rRNA sequence (1,388 bp) of the E. canis Kutahya was identical to that of Ehrlichia ovina from a sheep in Turkey and Venezuelan Dog Ehrlichia (VDE) and was closely related (99.9%) to that of type strain of E. canis, Oklahoma. The plasma of the dog infected with E. canis Kutahya was analyzed by Western blotting using the purified E. canis Oklahoma strain as antigen. The reactive antibody profiles of the dog infected with E. canis Kutahya was found to be similar to those of dogs infected with E. canis Oklahoma and VDE, suggesting the antigenic similarities among these strains. The findings in this study would help for a better understanding of epidemiology of canine ehrlichiosis. This is the first report of molecular detection and characterization of an ehrlichial agent in Turkey.     

Antigenic analysis of four species of the genus Ehrlichia by use of protein immunoblot

The antigenic profile of Ehrlichia canis, E risticii, E sennetsu, and E equi was investigated by the use of protein (western) immunoblot technique. Results of analysis of serum from acutely and chronically infected animals indicated that the 4 Ehrlichia species share a unique 25-kD polypeptide in addition to other peptides. Immune sera from dogs inoculated with E canis recognized a wide range of E canis polypeptide antigens, as determined by western blot analysis. A larger number of E sennetsu polypeptides were detected when homologous antiserum and antiserum to E equi were used. The latter antiserum did not recognize antigens of E canis or E risticii. Antisera to E canis, E risticii, and E sennetsu detected E equi antigens. Data indicate that a 25-kD protein is a common antigen among the species of the genus Ehrlichia and that the ascending order of abundance of immunodominant determinants in the 4 species of Ehrlichia studied would be: E risticii----E equi----E sennetsu----E canis. Implications of these findings for diagnosis of ehrlichial infections and prophylaxis are evident.     

West-to-east differences of Babesia canis canis prevalence in Dermacentor reticulatus ticks in Slovakia

Babesia canis canis is the most frequent causative agent of canine babesiosis in Central Europe, frequently causing severe disease. Recently, many new endemic foci of this disease have been reported from European countries. Growing incidence of canine babesiosis was recorded also in Slovakia during the last decade, from first cases in eastern Slovakia ten years ago to recent cases all over the south of the country. We have used nested PCR-RFLP method to study prevalence of B. c. canis in its natural tick vector Dermacentor reticulatus, collected at three geographically isolated lowland areas of southern Slovakia situated in the southeast, southwest, and west of Slovakia, respectively. The highest prevalence of B. c. canis was observed in D. reticulatus from eastern Slovakia (14.7%; n=327), whereas the prevalence in southwest was significantly lower (2.3%; n=1205). Notably, all 874 D. reticulatus ticks collected at Záhorská ní?ina lowland (W Slovakia) were B. c. canis-negative. Recorded differences in Babesia prevalence concurs well with the shift in incidence of clinical cases of canine babesiosis as observed by vet practitioners. Presented results revealed that eastern Slovakia represents an area of high risk of B. c. canis infection, whereas western areas of the country still remain Babesia canis-free.     

A rapid slide agglutination test for the serodiagnosis of Brucella canis infection that employs a variant (M-) organism as antigen

An improved antigen is described for use in the serodiagnosis of canine brucellosis. The novel antigen employs a less mucoid (M-) variant of B. canis. The replacement of Brucella ovis with B. canis (M-) cells as antigen in slide agglutination tests would increase accuracy by reducing the rate of false positive results from ca. 50% to ca. 10%. Rose bengal stained B. canis (M-) cells are slightly more sensitive than the commercially available Brucella ovis as slide test antigens. Both test procedures require brief (less than 1 min) reaction with 2-mercaptoethanol in order to reduce the rate of false positive reactions.     

1例猫假足菌肿的诊断与治疗

为加强对罕见疾病假足菌肿的认识，给兽医工作者在诊治相似病例时提供参考，本研究分别从临床症状、诊断过程及治疗方法对1例犬小孢子菌引起的猫假足菌肿的病例进行报道。采集患猫尾荐部样本进行细胞学检查、真菌培养及菌株质谱鉴定。细胞学涂片中可见大量的炎性细胞、真菌孢子和真菌菌丝，诊断为真菌性肉芽肿性炎症。基于细胞学的结果，取样进行真菌培养。对培养出的菌株染色镜检，发现有6个以上分隔的大分生孢子。最后，通过质谱鉴定确定该病原菌为犬小孢子菌。综合以上检查结果，该病例诊断为犬小孢子菌引起的猫假足菌肿。治疗采用外用药浴与口服抗真菌药物相结合的方法。治疗初期，患猫尾部病灶有好转但并不明显，后期持续恶化，并于首诊后7个月死亡，主要死亡原因不明。建议兽医工作者在遇到相似病例时引起重视，采用细胞学检查、真菌培养及菌株鉴定等方法进行综合诊断。在疾病早期使用手术切除与抗真菌药物相结合的方法进行治疗，以提高患病动物的存活率。     

Serological and bacteriological detection of Brucella canis infection of stray dogs in Moreno, Argentina

A focus of B. canis infection was identified in Moreno, Argentina, among the stray dog population by serologic methods and confirmed in a second survey which included cultural isolations. A counterimmunoelectrophoresis technique using a specific rough Brucella surface antigen was applied to the serodiagnosis of canine brucellosis. This method was found to be as effective as the mercaptoethanol tube agglutination test and the gel diffusion test in detecting B. canis antibodies in natural and experimentally infected animals. The results are discussed in terms of the diagnostic significance of the three tests employed.     

Relatedness of Streptococcus canis from canine streptococcal toxic shock syndrome and necrotizing fasciitis.

The emergence of streptococcal toxic shock syndrome (STSS) and necrotizing fasciitis (NF) in dogs caused by Streptococcus canis has been reported by our laboratory. Since clonal expansion is thought to be partially responsible for the spread of invasive strains of Streptococcus pyogenes in humans, the relatedness of 15 isolates of S. canis from canine STSS and/or NF was examined using pulsed field gel electrophoresis and biotyping; production of proteases and of a CAMP-like reaction were also examined. Only 2 of the 15 STSS and/or NF isolates were clonally related, suggesting that the emergence of canine STSS/NF is not the result of clonal expansion of one or more highly virulent strains of S. canis. All of the isolates produced proteases and demonstrated a CAMP-like reaction, which appear to be additional characteristics of S. canis.     

Scrotal and testical changes in canine brucellosis: a case report

An Irish Setter with scrotal enlargement had an agglutination titer to Brucella canis of greater than or equal to 1:200, but B canis was not cultured from the blood. Unusual findings included draining ulcers in the scrotum from which B canis was cultured, a necrotic testicle, and inflammation of the entire scotum. Other clinical and postmortem findings were as previously described in other cases of naturally occurring and experimentally induced canine brucellosis.     

A preliminary study to evaluate the immune responses induced by immunization of dogs with inactivated Ehrlichia canis organisms

Ehrlichia canis is an intracellular pathogen that causes canine monocytic ehrlichiosis. Although the role of antibody responses cannot be discounted, control of this intracellular pathogen is expected to be by cell mediated immune responses. The immune responses in dogs immunized with inactivated E. canis organisms in combination with Quil A were evaluated. Immunization provoked strong humoral and cellular immune responses, which were demonstrable by Western blotting and lymphocyte proliferation assays. By Western blotting antibodies to several immunodominant E. canis proteins were detected in serum from immunized dogs and antibody titres increased after each immunization. The complement of immunogenic proteins recognized by the antisera were similar to those recognized in serum from infected dogs. Upon challenge with live E. canis, rapid anamnestic humoral responses were detected in the serum of immunized dogs and primary antibody responses were detected in the serum from control dogs. Following immunization, a lymphocyte proliferative response (cellular immunity) was detected in peripheral blood mononuclear cells (PBMNs) of immunized dogs upon stimulation with E. canis antigens. These responses were absent from non-immunized control dogs until after infection with live E. canis, when antigen specific-lymphocyte proliferation responses were also detected in the PBMNs of the control dogs. It can be thus concluded that immunization against canine monocytic ehrlichiosis may be feasible. However, the immunization regimen needs to be optimized and a detailed investigation needs to be done to determine if this regimen can prevent development of acute and chronic disease.     

Serodiagnosis of Babesia gibsoni infection in dogs by an improved enzyme-linked immunosorbent assay with recombinant truncated P50

The surface antigen P50 of Babesia gibsoni is an important candidate for the development of a diagnostic reagent for canine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding truncated P50 (P50t) lacking a signal peptide and C-terminal hydrophobic regions were cloned and expressed in Escherichia coli as a fusion protein with glutathione S-transferase (GST). More than 90% portion of the GST-P50t was expressed as a soluble form, in contrast with GST-P50f (full-length), which was completely expressed as an insoluble form. This result indicates that removal of the hydrophobic signal peptide and C-terminus had dramatically improved its hydrophilicity. The purified GST-P50t was tested in an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to B. gibsoni in dogs. The ELISA with GST-P50t clearly differentiated between B. gibsoni-infected dog sera and uninfected dog sera. In addition, the ELISA detected no cross-reactivity with sera from dogs experimentally infected with the closely related parasites, B. canis canis, B. canis vogeli, and B. canis rossi. Field serum samples collected from dogs in Japan and China were examined for the diagnosis of B. gibsoni infection by using the ELISA. 14.5% (9/62), 5.8% (7/120), and 5.4% (2/37) of tested samples were positive for dogs from Okinawa, Yamaguchi, and Osaka prefectures, Japan, respectively. On the other hand, 4.8% (2/41) of tested samples were positive for dogs from Nanjing, China. These results suggest that the GST-P50t could be a reliable reagent for practical use in ELISA for the serodiagnosis of canine piroplasmosis caused by B. gibsoni.     

Immunohistochemical diagnosis of Neospora caninum in tissue sections

An avidin-biotin-peroxidase complex immunoperoxidase staining method was developed to detect Neospora caninum in formalin-fixed, paraffin-embedded tissue sections. Specific antiserum to N caninum was made in rabbits and used to probe tissues from dogs naturally and experimentally infected with N caninum. The test detected tachyzoites and bradyzoites of N caninum. A reaction was not observed to Toxoplasma gondii, Hammondia hammondi, Sarcocystis cruzi, S capricanis, S tenella, Besnoitia jellisoni, Caryospora bigenetica, Hepatazoon canis, Atoxoplasma sp, or the organism causing canine dermal coccidiosis. When antiserum made in rabbits to T gondii was used in the test, reaction to N caninum was not observed.     

Doxycycline clearance of experimentally induced chronic Ehrlichia canis infection in dogs

BACKGROUND: Ineffective clearance of Ehrlichia canis after doxycycline administration has been reported despite the fact that the recommended treatment for canine ehrlichiosis is doxycycline. The effectiveness of doxycycline in clearing E canis infection from the blood and tissues of dogs requires additional evaluation. HYPOTHESIS: Doxycycline (5 mg/kg PO q12h), administered for 4 weeks, will eliminate E canis infection from the blood and tissues of experimentally infected dogs. ANIMALS: Fifteen Walker hound-mixed breed dogs were inoculated subcutaneously with E canis-infected canine histiocytic cells 4 months before doxycycline treatment. METHODS: Four dogs were treated with doxycycline (5 mg/kg PO q12h for 3 weeks), 5 dogs were treated with doxycycline at the same dosage for 4 weeks, and 5 control dogs were not treated. Dexamethasone (0.4 mg/kg i.v.) was given after treatment to precipitate recrudescence of any remaining E canis organisms. Platelet counts, anti-E canis immunofluorescent antibodies, and polymerase chain reaction (PCR) detection of E canis deoxyribonucleic acid (DNA) in blood and tissues were evaluated. RESULTS: E canis DNA was not detected in the blood and tissues of doxycycline-treated dogs after treatment. Platelet counts were within reference intervals, and E canis antibodies decreased. Spontaneous clearance of E canis infection occurred in 2 of 5 control dogs. Three control dogs had E canis DNA detected in blood and tissues, platelet counts remained low or within the reference interval, and E canis antibodies remained high. CONCLUSIONS AND CLINICAL IMPORTANCE: As administered in this study, doxycycline cleared E canis from the blood and tissues of experimentally infected dogs.     

Virulence of Streptococcus canis from canine streptococcal toxic shock syndrome and necrotizing fasciitis

The recent recognition of streptococcal toxic shock syndrome (STSS) and necrotizing fasciitis (NF) in dogs caused by Streptococcus canis highlights our lack of knowledge regarding the mechanisms of virulence of this organism. Fifteen isolates of S. canis from cases of canine STSS and/or NF were examined for the presence of 10 Streptococcus pyogenes-associated virulence genes by Southern hybridizations using gene probes generated by PCR. The isolates lacked DNA with homology to eight of the 10 gene probes (speA, speB, speC, mf, ssa, scp, hasA, ska) under low stringency conditions. Thirteen and 15 of 15 isolates hybridized with streptolysin O and M protein gene probes, respectively. Twelve of 15 S. canis isolates were resistant to phagocytosis in canine blood. Electron microscopy revealed the presence of proteinaceous cell surface fibrillae. These results suggest that S. canis possesses M proteins and encodes streptolysin O, but lacks some of the other recognized virulence genes with significant homology to those in S. pyogenes.     

Clinical Sarcocystis neurona, Sarcocystis canis, Toxoplasma gondii, and Neospora caninum infections in dogs

Sarcocystis neurona, Sarcocystis canis, Toxoplasma gondii, and Neospora caninum are related apicomplexans that can cause systemic illness in many species of animals, including dogs. We investigated one breeder's 25 Basset Hounds for these infections. In addition, tissues from dogs and other non-canine hosts previously reported as S. canis infections were studied retrospectively. Schizonts resembling those of S. neurona, and recognized by polyclonal rabbit anti-S. neurona antibodies, were found in six of eight retrospective cases, as well as in two additional dogs (one Basset Hound, one Springer Spaniel) not previously reported. S. neurona schizonts were found in several tissues including the central nervous system, lungs, and kidneys. Fatal toxoplasmosis was diagnosed in an adult dog, and neosporosis was diagnosed in an adult and a pup related to the one diagnosed with S. neurona. No serological reactivity to S. neurona antibodies occurred when S. canis-like liver schizonts were retrospectively assayed from two dogs, a dolphin, a sea lion, a horse, a chinchilla, a black or either of two polar bears. Sequencing conserved (18S) and variable (ITS-1) portions of nuclear ribosomal DNA isolated from the schizont-laden liver of a polar bear distinguished it from all previously characterized species of Sarcocystis. We take this genetic signature as provisionally representative of S. canis, an assumption that should be tested with future sequencing of similar liver infections in other mammalian hosts. These findings further extend the uncharacteristically broad intermediate host range for S. neurona, which also causes a neurologic disease in cats, mink, raccoons, skunks, Pacific harbor seals, ponies, zebras, lynxes, and sea otters. Further work is necessary to delineate the causative agent(s) of other cases of canine sarcocystosis, and in particular to specify the attributes of S. canis, which corresponds morphologically to infections reported from wide range of terrestrial and marine mammals.