Development of an enzyme-linked immunosorbant assay (ELISA) for the serodiagnosis of canine dermatophytosis caused by Microsporum canis

Abstract In dogs, dermatophytosis should be considered in any case of alopecic, papular or pustular lesion. The aim of this study was to develop an enzyme-linked immunosorbant assay (ELISA) as an aid in the diagnosis of canine dermatophytosis. The antigen used was a whole fungal extract obtained from an isolate of Microsporum canis cultured on a liquid medium from the parasitized hair of a cat with patches of alopecia. To assess the ELISA performances, sera from 18 dogs with dermatophytosis caused by M. canis (group A, n = 18), 20 dogs with skin diseases other than dermatophytosis and 22 healthy dogs (group B, n = 42) were tested. Four further animals were tested: three with dermatophytosis caused by M. gypseum and one by T. mentagrophytes. A significant difference (P < 0.01, Wilcoxon's test, w = 364) was found between IgG-specific levels of sera of recently M. canis-infected dogs (infection < 15 days) and controls (although three dogs had negative titres at this stage). A highly significant difference (P < 0.001, w = 462) was noted between controls and dogs with infection of longer duration (> 30 days). All dogs had positive titres at this stage. A highly significant correlation (P < 0.001, Spearman's test, rho = 0.86) between duration of infection and IgG concentration was noted. The test has good sensitivity (83.3%) and high specificity (95.2%) but some dogs retained positive titres after elimination of infection. The sensitivity is higher than that of direct microscopic hair examination and similar to that of fungal culture with DTM (dermatophyte test medium).     

Efficacy of itraconazole as a combined continuous/pulse therapy in feline dermatophytosis: preliminary results in nine cases

This study evaluated the efficacy of itraconazole as a combined continuous/pulse therapy for feline dermatophytosis. Nine cats with dermatophytosis caused by Microsporum canis were treated with itraconazole at 10 mg kg(-1) orally once daily for 28 days and then on an alternate week regimen (1 week off, 1 week on) at the same dosage. Cats were re-evaluated by physical examination and fungal culture at days 28, 42, 56 and 70 if necessary. Treatment was stopped when two consecutive negative fungal cultures were obtained. Eight cats were cured after 56 days, with two negative cultures obtained at days 28 and 42. In one case, a positive culture was obtained at day 28, but negative cultures were achieved at days 42 and 56. This protocol appears to be effective in the treatment of feline dermatophytosis, although these preliminary results should be confirmed by a controlled study.     

Safety and immunologic effects after inoculation of inactivated and combined live-inactivated dermatophytosis vaccines in cats

OBJECTIVE: To determine antidermatophyte immunologic effects of an experimental combined live-inactivated dermatophytosis vaccine (CLIDV) and a commercial inactivated dermatophytosis vaccine (IDV) in cats and to evaluate adverse effects associated with administration of these vaccines. ANIMALS: 20 healthy juvenile domestic shorthair cats. PROCEDURE: Cats were injected with 2 doses of CLIDV at the standard dosage or 1 dose of CLIDV at 10 times the standard dosage; IDV was administered at the manufacturer-recommended dosage. Cats were observed for illness and reactions at inoculation sites. Periodically, samples were obtained for fungal culture, lymphocyte blastogenesis test (LBT) as an indicator of cell-mediated immunity against dermatophyte antigens, and antidermatophyte IgG titers. Following vaccination, cats were challenge-exposed by topical application of Microsporum canis macroconidia and examined weekly for clinical signs of dermatophytosis. RESULTS: of 10 cats given CLIDV developed focal crusts at the injection site that resolved without treatment; these were areas of dermatophyte infection with the vaccine strain. Antidermatophyte IgG titers increased significantly with all vaccination protocols. Cellular immunity against M canis increased slightly and variably during the vaccination period and did not differ significantly between vaccinated and control cats. All cats developed dermatophyte infection after challenge exposure. Vaccination with CLIDV or IDV was associated with slightly reduced severity of initial infection. CONCLUSIONS AND CLINICAL RELEVANCE: Noculation with IDV or CLIDV did not provide prophylactic immunity against topical challenge exposure with M canis. Inoculation with either vaccine did not provide a more rapid cure of an established infection.     

Efficacy of pre-treatment with lufenuron for the prevention of Microsporum canis infection in a feline direct topical challenge model

Oral lufenuron is reportedly an effective treatment for some cats with dermatophytosis. The purpose of this study was to determine if lufenuron, when used as a pre-treatment prior to challenge exposure, would be protective against the development of infection after the direct topical application of fungal macrocondia (Microsporum canis spores). Three groups (n = 6/group) of juvenile cats were treated with either monthly oral lufenuron (30 or 133 mg/kg) or placebo. After 2 months of treatment, kittens were challenged using 10(5)Microsporum canis spores applied to the skin under occlusion. Cats were examined weekly and the following data collected: Wood's lamp examination; scoring for scale/crust, erythema and induration; lesion size; and the development of satellite lesions. Fungal cultures were performed bi-weekly. All cats became infected; the infections progressed, and then regressed, in a similar fashion in all groups. There were no consistent statistically significant differences in weekly infection scores between treated and untreated cats throughout the study. Treated cats did not recover faster than untreated cats. We conclude that oral lufenuron at the dosing schedule and conditions used in this study did not prevent dermatophytosis or alter the course of infection by direct topical challenge.     

Isolation of fungal flora from the hair coats of shelter cats in the Pacific coastal USA

Two-hundred shelter cats from the Pacific western coastal USA were sampled in four different geographical regions to determine the fungal organisms most commonly found on the hair coat and the prevalence of these organisms. Data on the cats' health, age, hair coat length, gender, and geographical location were collected and analysed. The overall prevalence of dermatophytosis was 5.5% (11 of 200 cats), with Microsporum canis isolated in 90.9% (10 of 11) of the samples from positive cats. This was a lower isolation rate or prevalence of dermatophytes than previous studies conducted on shelter cats in other regions of the USA. Ten of 11 of the cats were lesion free (either subclinical infection or mechanical carriage). Cats in the Los Angeles, California area ( P  = 0.001) and neutered male cats ( P  = 0.047) had a higher prevalence of a positive dermatophyte culture. The numbers and types of saprophytes isolated from cats in this study were found to be consistent with previous feline reports in the USA and with an equine study previously conducted in this area.     

Frequency of yeasts and dermatophytes from healthy and diseased dogs.

The aim of this study was to investigate the presence of dermatophytes and yeasts in healthy and diseased dogs. A total of 633 samples were collected from 26 healthy animals (104 samples), 131 with dermatitis (343 samples), 74 with otitis (148 samples), and 19 with ocular diseases (38 samples). Cultures from healthy animals were positive for Malassezia pachydermatis in 13.5% (7/52) of samples from skin, 42.3% (11/26) from ear, and 3.8% (1/26) from eye. Fungal growth was observed in 20.4% (70/343) samples from animals with dermatitis. Microsporum canis was the most isolated fungus (n = 39), followed by M. pachydermatis (n = 30) and Malassezia sp. (n = 3). Of the 148 samples from dogs with otitis, 90 (60.8%) were positive for M. pachydermatis, and of the clinical specimens from the conjunctiva of animals with ophthalmic disease, 2.6% (1/38) presented positive cultures for M. pachydermatis. Only 14.3% (2/14) of the positive cultures for M. pachydermatis and 40.9% (9/22) of those for M. canis were positive in the direct exam. Direct exams were positive in 84.3% (70/83) of the culture positive samples from affected ears of dogs with otitis. Malassezia pachydermatis may act as an aggravating factor in the occurrence of cutaneous diseases, or the isolation of M. canis may be associated with the onset of dermatophytosis. Fungal culture, rather than microscopic examination, should be used as the definitive diagnostic test for dermatomycoses and otitis.     

Evaluation of topically applied enilconazole for the treatment of dermatophytosis in a Persian cattery

Many Persian catteries have long-standing dermatophyte infections and are particularly difficult to treat. Enilconazole is a topical antifungal agent that has demonstrated good efficacy in recent studies. Twenty-two Persian cats naturally infected with Microsporum canis in a breeding cattery were treated with topical 0.2% enilconazole and monitored for 180 days. The treatments were repeated every 3 days for a total of eight applications. All the cats improved clinically and became culture negative by day 28. By day 180, four cats had developed clinical dermatophytosis and all cats had positive fungal cultures. In this study, topical 0.2% enilconazole was generally well tolerated but may have caused hypersalivation, idiopathic muscle weakness and slightly elevated serum alanine aminotransferase (ALT) concentrations. This study suggests that enilconazole may be used safely with little risk to the young, aged and gravid animals.     

Isolation of Microsporum canis from the hair coat of pet dogs and cats belonging to owners diagnosed with M. canis tinea corporis

Microsporum canis has been frequently isolated from human cases of tinea capitis and tinea corporis. The infection may be acquired from infected animals with cutaneous lesions but also from asymptomatic carriers or from the environment. As asymptomatic M. canis carriers are considered to be a critical factor in the epidemiology of dermatophytosis in humans, this study investigated the relationship between the presence of dermatophytes on the hair coats of dogs and cats without cutaneous lesions and the occurrence of the disease in their respective owners. A total of 136 dogs and 248 cats were sampled from January 1999 to January 2005. Seventy-eight animals (22 dogs and 56 cats) belonged to individuals affected by tinea corporis caused by M. canis and 306 (114 dogs and 192 cats) to individuals without dermatophytosis. Age, sex, breed, habitat and season were recorded for each animal and examined as potential risk factors. Dermatophytes were isolated from 20.5% of the dogs and 28.2% of the cats. Microsporum canis was isolated from 36.4% of dogs cohabiting with owners diagnosed with tinea corporis but it was never isolated from dogs whose owners had no lesions. By contrast, M. canis was isolated from 53.6% of cats cohabiting with owners diagnosed with tinea corporis and from 14.6% of cats whose owners had no signs of the disease. These results clearly indicate that both cats and dogs should be considered as a major source of pathogenic dermatophytes for humans even when they do not present clinical signs of dermatophytosis.     

Dermatophytosis and papular eosinophilic/mastocytic dermatitis (urticaria pigmentosa-like dermatitis) in three Devon Rex cats

PRESENTING SIGNS: Three Devon Rex cats were presented with multiple erythematous papules, occasionally associated with crusting and hyperpigmentation, with a linear distribution on the head, neck, chest and abdomen. One cat also had multifocal alopecia with hyperpigmentation on the dorsum. DIAGNOSIS AND TREATMENT: Clinical and histopathological features were suggestive of papular eosinophilic/mastocytic dermatitis (urticaria pigmentosa-like dermatitis). In all cases, dermatophytosis was diagnosed: in cases 1 and 2 there was histopathological evidence of dermatophytosis, while fungal culture was positive for Microsporum canis in cases 2 and 3. In all cats, lesions disappeared following antifungal treatment. CLINICAL SIGNIFICANCE: Papular eosinophilic/mastocytic dermatitis in Devon Rex cats may represent either an atypical presentation of dermatophytosis or a clinical and histological reaction pattern to various diseases, including dermatophytosis and allergic diseases. Clinical differentiation is crucial as there are important implications regarding treatment and, in particular, the use of glucocorticoids, which are contraindicated in cases of dermatophytosis.     

Drug efficacy of terbinafine hydrochloride (Lamisil) during oral treatment of cats,experimentally infected with Microsporum canis

Cats represent a primary source of Microsporum canis infections in humans. Terbinafine hydrochloride (Lamisil) is commonly used in the treatment of microsporosis in humans as its fungicidal action permits short periods of treatment. The aim of the present study was to estimate the efficacy of the drug in cats. Nine cats were experimentally infected with M. canis and treated with terbinafine hydrochloride at a dose of 10-20 mg/kg (once daily, SID; low-dose group, LDG). Another nine cats were similarly infected and treated with 30-40 mg/kg SID (high-dose group, HDG) and a further nine cats were also infected and left untreated (control group, CG). The general condition of the cats was observed daily and their clinical symptoms evaluated weekly. The cats recovery was monitored using the Wood's lamp illumination test and microscopic and fungal culture examinations. The general condition of the cats during the study was good. The cure rates of the LDG were not significantly different from the CG at any period during the treatment. However, the HDG cure rates differed significantly from the other two groups. After 109 days of treatment, when all nine cats of the HDG were healed, seven cats of the LDG and all the cats in the CG were still M. canis-positive. This study shows that dosages of 10-20 mg/kg SID of terbinafine hydrochloride are not sufficient to terminate an experimental M. canis infection in cats within an acceptable period of time. Terbinafine hydrochloride can be used to treat dermatophytosis in cats, but a higher dosage, 30-40 mg/kg SID, should be used to achieve a cure.     

P-15 Identification of Malassezia spp. isolated from a cat and cattle, including a new species, M. nana

Preliminary studies showed that lufenuron inhibits chitin synthesis, a dermatophyte cell wall constituent, and may be effective in the treatment of dermatophytosis. Our purpose was to evaluate the efficacy of lufenuron in the treatment of feline dermatophytosis. Forty-six cats (Persians and mixed-breed cats from 1-month to 4-years old) naturally infected with Microsporum canis were included in this study. Fifteen cats were treated isolated in cages in the veterinary hospital and 31 were treated in their home environment (some with access to the outdoors). Dermatophyte skin lesions were seen in 29 animals while 17 other cats were asymptomatic carriers. Wood's lamp, direct microscopic examination of hairs, fungal culture and skin biopsies were used for the diagnosis. Affected cats and all in-contact animals received lufenuron at a dose of 120 mg/kg every 21 days for four treatments. Of the 29 symptomatic cats treated with lufenuron, 70% recovered within 21 days and 28% within 42 days of initiation of therapy. One cat had only partial recovery and another was euthanized. Negative fungal culture was recorded only after the fourth dose of lufenuron in 98% of affected cats and 100% of asymptomatic carriers. There was no difference in clinical response to lufenuron between the cats treated in their home environment and those treated in the veterinary hospital. Side effects were not observed, thus the drug proved to be safe and effective for the treatment of dermatophytosis.
Funding: Novartis.     

Pilot study: prevalence of positive aeroallergen reactions in 10 cats with small-airway disease without concurrent skin disease

The purpose of this study was to determine the prevalence of positive allergen reactions in cats with small-airway disease (i.e. 'feline asthma', 'feline allergic bronchitis', 'feline bronchial disease'). Intradermal skin tests (IDT) and serum immunoglobulin E (IgE) tests were performed in 10 cats with idiopathic small-airway disease and in 10 normal cats without a history of respiratory disease. None of the cats had a history of skin disease or clinical signs of skin disease at the time of testing. Significantly more individual positive allergen reactions were found on serum IgE tests than on IDT in both groups of cats. Affected cats had significantly more individual positive allergen reactions on both tests than unaffected cats. Both IDT and serum IgE tests resulted in more individual positive allergen reactions to weeds, trees, grasses, and/or moulds in affected cats than in normal cats. Significantly more positive allergen reactions to house dust mites were found in affected compared to non-affected cats by IDT but not by serum IgE testing. One unexpected obstacle to inclusion of more affected cats in the study was the concurrent presence or history of suspect or known allergic skin disease. Concurrent allergic skin disease has not been reported in association with small-airway disease in cats. The increased prevalence of individual positive allergen reactions in affected cats may be due to increased immunological reactivity in these cats. Further studies are needed to answer this question and to determine what role, if any, aeroallergens have in the pathogenesis of this complex feline disease.     

Effects of lufenuron treatment in cats on the establishment and course of Microsporum canis infection following exposure to infected cats

OBJECTIVE: To determine effects of lufenuron treatment in cats on the establishment and course of Microsporum canis infection following exposure to infected cats. DESIGN: Experimental trial. ANIMALS: 24 healthy juvenile domestic shorthair cats. PROCEDURE: 8 cats were given lufenuron PO (133 mg/cat/mo, equivalent to a dose of 100 to 130 mg/kg [45 to 59 mg/lb] at the beginning of the study and 25 to 35 mg/kg [11 to 16 mg/lb] at the end of the study), and 8 were given lufenuron SC (40 mg every 6 months). The remaining 8 were used as untreated control cats. After 4 months, cats were challenged by the introduction of cats with mild, experimentally induced M canis infection into the rooms where cats were housed. Extent of resulting infections in the na?ve cats was monitored for 22 weeks by physical examination and fungal culture. RESULTS: All lufenuron-treated and control cats became infected with M canis. Cats treated with lufenuron had significantly lower infection scores, compared with control cats, during the early weeks following exposure, and there was a more prolonged initial progression phase of the infection. Once infections reached peak intensity, they resolved over similar periods in lufenuron-treated and control cats. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that oral or SC administration of lufenuron to cats, at the dosages used and under the conditions of this study, did not prevent establishment of dermatophytosis following exposure to infected cats. Infection was established more slowly among cats treated with lufenuron, but once established, infection resolved in approximately the same amount of time in lufenuron-treated as in control cats.     

Environmental detection of Microsporum canis arthrospores in the households of infected cats and dogs

Microsporum canis is the dermatophyte most frequently recovered from canine and feline ringworm cases. The household environment can be contaminated both by symptomatic animals and through asymptomatic M canis carriage, resulting in a potential human health risk. The load of M canis arthrospores was determined in households harbouring infected pets, in order to evaluate the infectivity of the animals versus the environment. The environments inhabited by 30 symptomatic animals (21 cats and 9 dogs) infected by M canis were examined by sampling both surfaces and indoor air. The surfaces were examined by means of contact plates; the air sampling was performed with a Sas super-100 AIR SAMPLER (PBI, Italy). Environmental contamination was detected in all households with cats, while only four out of nine houses harbouring dogs were found positive. The frequence of isolation in each sampling, and the results in terms of colony forming units per plate in the different houses appeared to be quite homogeneous. Heavily infected environments harboured kittens only. Infected owners were observed in eight households, in all of which at least one infected cat was present. No history of human dermatophytosis in households harbouring dogs was found. On the basis of our results, infected cats appear to cause substantial environmental contamination, and provoke a substantial presence of viable airborne fungal elements. Dogs seem to be of lower importance in the spread of M CANIS: they contaminated surfaces, but they never contaminated the air. The results of this study confirm the potential leading role of the feline species in the environmental spread of M canis.     

Use of a sustained release preparation of clotrimazole to treat dermatophytosis in a siamang (Hylobates syndactylus).

In November 2004, an adult male siamang (Hylobates syndactylus) from The Tisch Family Zoological Gardens-Jerusalem Biblical Zoo (Israel) presented with skin lesions on various body parts. Lesions consisted of alopecia and dry, crusty areas of hyperkeratosis. A diagnosis of dermatophytosis due to Microsporum canis was determined by fungal culture of skin scraping taken from the edge of several lesions. Treatment with various oral and topical antifungal agents such as griseofluvin, itraconozole, and lufenuron resulted in the resolution of most lesions and a decrease in size of the single remaining lesion, which continued to be culture positive for M. canis. The animal was anesthetized and an experimental sustained-release clotrimazole varnish was painted directly on the lesion. Initially there was no change in the lesion, and 2 months later a slightly altered formula was applied under anesthesia. One month later, the lesion began to reduce in size; 3 months after the start of treatment, although 2 years after the onset of clinical signs, the lesion resolved. Minimizing the number of treatments is always an advantage when dealing with exotic animals or zoological collections.     

Generalized Microsporum canis dermatophytosis in six Yorkshire terrier dogs

Six Yorkshire terrier dogs with generalized, chronic dermatophytosis caused by Microsporum canis were seen over a 3-year period. Specific tests showed that they also had concurrent leishmaniosis (four cases), leishmaniosis and ehrlichiosis (one case) or diabetes mellitus (one case). Although specific therapy for these infectious diseases was instituted and the dogs were treated systemically and topically with appropriate antifungal drugs, only partial clinical resolution of the dermatophytosis was achieved. M. canis infection resolved in the dog with diabetes mellitus after stabilizing the diabetes mellitus. Although immunological studies were not performed in these cases, it is theorized that the immune disregulation caused by leishmaniosis, ehrlichiosis or diabetes mellitus may have favoured generalization of the infection and prevented favourable responses to appropriate treatment of the M. canis infection.     

Cortisol levels in cats’ hair in presence or absence of Microsporum canis infection

The purpose of this work was to perform a preliminary screening in the domestic cat to assess the concentration of cortisol in hairs by radioimmunoassay technique (RIA) in presence or absence of Microsporum canis infections. A total of 245 cats (7 with cutaneous lesions referable to dermatophytosis and 238 apparently healthy) coming from 14 shelters were examined. M. canis was isolated in 126 (51.4%) cats. The cortisol levels were significantly higher in cats with lesions or without lesions but with a high number of colonies in the plates (⩾10 CFU) than in cats negative or with a lower number of colonies. The results obtained seem to highlight that chronic high levels of cortisol in cats could possibly promote the dermatophytes infections. Furthermore, in High-CFU asymptomatic cats, it could be present a state of infectious, and they, therefore, represents not a simple mechanical carrier.     

P-12 Comparative study of griseofulvin and terbinafine therapy in the treatment of canine and feline dermatophytosis

Diagnosis of Microsporum canis infection is a challenge in cats with suspected asymptomatic carriage. The aim of this study was to compare the carpet method and the toothbrush method of sample collection for dermatophyte culture. The study was conducted on apparently healthy cats in a chronically infected cattery. Sampling was performed with both a sterilized piece of carpet and a toothbrush applied to the entire body surface. Samples were inoculated on Sabouraud's media (with chloramphenicol and cycloheximide). Multiple (6) applications with the toothbrush were necessary to obtain a similar surface area of inoculation. Cultures were incubated at 27°C, then examined from day 5 to 21 for the presence and number of colonies of M. canis and other fungi. Some samples were inoculated twice (initially negative or with a low number of M.canis ). A total of 112 cultures were performed (14 duplicates) from 44 cats. On day 21, infection was detected in 23 cats (54.7%): 20 (87%) with the carpet method and 21 (91%) with the toothbrush method. None of the replicated cultures allowed the detection of new cases but three were subsequently negative. In eight cases, only one to six colonies of M. canis were obtained. Both methods used are easy to perform on cats and have a similar sensitivity of approximately 90%. The carpet method is, however, much less expensive, easier to prepare, mail, store and inoculate. The lack of growth in some duplicate cultures (initial development of one or two colonies) may explain the limits of diagnosis of infection and justify the need for multiple inoculations of agar with the toothbrush method.
Funding: Self-funded.     

Use of lime sulphur and itraconazole to treat shelter cats naturally infected with Microsporum canis in an annex facility: an open field trial

Dermatophytosis is the most common contagious and infectious skin disease of cats. It is of particular importance in animal shelters because it is a known zoonosis, highly contagious, and easily transmitted. In this open clinical trial, 58 cats with confirmed Microsporum canis dermatophytosis and 32 uninfected bonded pairs or littermates were treated with a combination of 21 days of oral itraconazole (10 mg kg(-1)) and twice weekly lime sulphur rinses until cured. Cats were not clipped in this treatment programme. Fungal cultures were obtained once weekly on all cats, and cats were considered cured when they had two consecutive negative weekly fungal cultures. Cats were held in the facility and received continued topical treatment until the fungal cultures were finalized. None of the cats developed oral ulcerations as a result of grooming the lime sulphur rinses. Oral ulcerations only developed in cats with clinical signs associated with upper respiratory disease. None of the uninfected cats living in contact with infected cats became culture positive or developed skin lesions. When data were examined retrospectively and the number of days to finalize the cultures was subtracted (21 days) from the total number of days the cats were housed in the annex, the mean number of days of treatment required for cure was 18.4 +/- 9.5 SEM (range 10-49 days). Cats with more severe infections required longer therapy. In this shelter, the combination of oral itraconazole and topical lime sulphur rinses for the treatment of dermatophytosis was effective and safe.     

Presence of Mycoplasma haemofelis,Mycoplasma haemominutum and piroplasmids in cats from southern Europe: a molecular study

Clinical symptoms produced by Mycoplasma spp. and piroplasmids in cats are sometimes similar. Diagnosis of these pathogens is difficult by microscopic procedures and molecular methods have been used as an alternative. We present in this work, the development of new molecular procedures for diagnosis of the aforementioned organisms, together with a molecular characterization of isolates found in southern European cats.A single PCR-RFLP procedure was designed for diagnosis of Mycoplasma spp. and a seminested PCR-RFLP was designed for diagnosis of piroplasmids. The 16S or 18S rRNA genes of isolates found in clinical samples were partially sequenced in all positive cases.Mycoplasma spp. was detected in 9 (30%) out of 30 symptomatic cats from Spain. Sequencing indicated that 66.6% of these isolates can be ascribed to Mycoplasma haemofelis and only 33.3% to Mycoplasma haemominutum. Partial 16S rRNA sequences obtained in Spanish isolates were very similar to those previously published from the UK and the USA.The presence of piroplasmids (Babesia and Theileria spp.) was studied in 16 cats from Spain (n=13) and Portugal (n=3). Animals analyzed were 10 cats with immunosuppressive viral infection (either FeLV or FIV), 5 asymptomatic cats and 1 cat with Babesia-compatible symptoms. Asymptomatic cats were all PCR-negative. Partial sequencing of 18S rRNA gene demonstrated that the Babesia-symptomatic cat was infected with Babesia canis canis whereas 3 (30%) out of the 10 cats with immunosuppressive viral infection were coinfected with piroplasmids (1 with B. canis canis, 1 with Theileria annae, and 1 with B. canis canis and T. annae both).