Tritrichomonas foetus and Mycoplasma felis coinfection in the upper respiratory tract of a cat with chronic purulent nasal discharge.

A 5-year-old indoor male neutered Siamese cat was presented with clinical signs of sneezing and chronic bilateral purulent nasal discharge. Multiple nasal cavity swabs were submitted for bacterial cultures, Mycoplasma felis-DNA qPCR, and cytology. M felisqPCR was positive and cytomorphologic diagnosis was severe, acute, purulent, rhinitis with intralesional protozoal microorganisms consistent with a Trichomonas spp. Nested PCR (nPCR) confirmed the diagnosis of Tritrichomonas foetus. Systemic therapy with doxycycline for M felis and metronidazole for T foetus was started with remission of clinical signs within 2 weeks; however, symptoms relapsed shortly after therapy was discontinued. This study represents the first documented case of T foetus associated with chronic nasal discharge in a cat, which supports the hypothesis that T foetus can live in the nasal cavity. It is also the first reported case of M felis and T foetus coinfection, which indicates that with mycoplasmal feline upper respiratory tract infections, T foetus should be considered as a coinfecting agent.

Open-field study comparing an essential oil-based shampoo with miconazole/chlorhexidine for haircoat disinfection in cats with spontaneous microsporiasis.

Objectives The goal of the present study was to compare the antifungal efficacy of an essential oil (EO) shampoo proven to be effective against Microsporum canis with miconazole/chlorhexidine for topical haircoat disinfection in cats treated concurrently with oral itraconazole. Methods Cats received treatment with oral itraconazole (Itrafungol) at a dose of 5 mg/kg/day pulse administration for 1 week, every 2 weeks for at least 6 weeks and were washed twice a week with a neutral shampoo with added EOs of Thymus serpyllum (2%), Origanum vulgare and Rosmarinus officinalis (5% each) for the period of systemic treatment. This protocol was compared with a conventional treatment (oral itraconazole + 2% miconazole/2% chlorhexidine shampoo). Results The treatment was well tolerated and adverse effects were not recorded. All cats were clinically negative at week 11. With respect to animals with extensive lesions, the speed of resolution was higher in cats with focal lesions. The animals showing diffuse lesions required more than a course of treatment to achieve a mycological cure. There was no significant difference between the number of weeks to obtain mycological cure for cats treated with EOs and animals treated conventionally. Conclusions and relevance The treatment appeared to be effective and well appreciated by the owners. The use of shampoo with the added EOs of T serpyllum, O vulgare and R officinalis would seem an interesting, natural alternative to conventional topical treatment.

In Vitro Activity of Twenty Commercially Available, Plant-Derived Essential Oils against Selected Dermatophyte Species.

The in vitro activity of twenty chemically defined essential oils (EOs) obtained from Boswellia sacra, Citrus bergamia, C. limon, C. medica, Cinnamomum zeylanicum, Eucalyptus globulus, Foeniculum vulgare, Helichrysum italicum, Illicium verum, Litsea cubeba, Mentha spicata, Myrtus communis, Ocimum basilicum, Origanum majorana, O. vulgare, Pelargonium graveolens, Rosmarinus officinalis, Santalum album, Satureja montana, and Thymus serpyllum was assayed against clinical animal isolates of Microsporum canis, Trichophyton mentagrophytes, T. erinacei, T. terrestre and Microsporum gypseum, main causative agents of zoonotic and/or environmental dermatophytoses in humans. Single main components present in high amounts in such EOs were also tested. Different dermatophyte species showed remarkable differences in sensitivity. In general, more effective EOs were T. serpyllum (MIC range 0.025%-0.25%), O. vulgare (MIC range 0.025%-0.5%) and L. cubeba (MIC range 0.025%-1.5%). F. vulgare showed a moderate efficacy against geophilic species such as M gypseum and T terrestre. Among single main components tested, neral was the most active (MIC and MFC values 5 0.25%). The results of the present study seem to be promising for an in vivo use of some assayed EOs.

Susceptibility of Microsporum canis arthrospores to a mixture of chemically defined essential oils: a perspective for environmental decontamination.

The zoophilic dermatophyte Microsporum canis has cats as natural reservoir, but it is able to infect a wide range of hosts, including humans, where different clinical features of the so-called ringworm dermatophytosis have been described. Human infections are increasingly been reported in Mediterranean countries. A reliable control program against M. canis infection in cats should include an antifungal treatment of both the infected animals and their living environment. In this article, a herbal mixture composed of chemically defined essential oils (EOs) of Litsea cubeba (1%), Illicium verum, Foeniculum vulgare, and Pelargonium graveolens (0.5% each) was formulated and its antifungal activity assessed against M. canis arthrospores which represent the infective environmental stage of M. canis. Single compounds present in higher amounts in the mixture were also separately tested in vitro. Litsea cubeba and P. graveolens EOs were most effective (minimum inhibitory concentration (MIC) 0.5%), followed by EOs of I. verum (MIC 2%) and F. vulgare (MIC 2.5%). Minimum fungicidal concentrations (MFC) values were 0.75% (L. cubeba), 1.5% (P. graveolens), 2.5% (I. verum) and 3% (F. vulgare). MIC and MFC values of the mixture were 0.25% and 0.5%, respectively. The daily spray of the mixture (200 µL) directly onto infected hairs inhibited fungal growth from the fourth day onwards. The compounds present in higher amounts exhibited variable antimycotic activity, with MIC values ranging from >10% (limonene) to 0.1% (geranial and neral). Thus, the mixture showed a good antifungal activity against arthrospores present in infected hairs. These results are promising for a further application of the mixture as an alternative tool or as an adjuvant in the environmental control of feline microsporosis.

A retrospective molecular study of select intestinal protozoa in healthy pet cats from Italy.

The feline gut can harbour a number of protozoan parasites. Recent genetic studies have highlighted new epidemiological findings about species of Cryptosporidium, assemblages of Giardia duodenalis and Toxoplasma gondii. Furthermore, epidemiological studies suggest the occurrence of Tritrichomonas foetus in cats is on the increase worldwide. The prevalence of selected intestinal protozoa was determined by PCR using DNA previously extracted from the faeces of 146 privately owned healthy cats from Italy. Molecular genotyping on T gondii, G duodenalis and Cryptosporidium DNA was achieved. PCR assays were positive in 32 (22.9%) samples. Three animals (2.0%) were positive for T foetus and Cryptosporidium DNA, 15 specimens (10.3%) were positive for T gondii and 11 (7.5%) for G duodenalis. Co-infections were never observed. Results of the typing analysis allowed the identification of Cryptosporidium felis in all cases. The specimens positive for T gondii hinted at clonal genotype I (n = 7), genotype II (n = 1) and genotype III (n = 7). The G duodenalis isolates were referable to assemblages F (n = 9) and C (n = 2). In conclusion, the results obtained in this study add to the literature regarding the epidemiology of these parasites by confirming their presence in the faeces of healthy pet cats.

Detection and genotyping of Toxoplasma gondii DNA in the blood and milk of naturally infected donkeys (Equus asinus).

BACKGROUND: Toxoplasma gondii is a worldwide zoonotic protozoan. Consumption of raw milk from infected animals is considered a risk factor for acquiring toxoplasmosis in humans. Recently, donkey milk has been indicated for therapeutic and nutritional purposes and T. gondii infection is common in donkeys. The purpose of the present paper was to detect the presence of parasite DNA in milk of T. gondii positive donkeys. FINDINGS: Antibodies to T. gondii were found in 11 out of 44 healthy lactating donkeys by IFAT. T. gondii DNA was detected by PCR in blood of 6 and milk of 3 seropositive jennies. Results of limited RFLP-PCR genotyping indicated the presence of T. gondii genotype II or III, commonly found in Europe. CONCLUSIONS: The occurrence of T. gondii DNA in milk suggests that the consumption of raw milk from seropositive donkeys could be a potential source of human infection.

Conjunctival bacterial and fungal flora in clinically normal sheep.

OBJECTIVES: The aim was to identify conjunctival bacterial and fungal flora in clinically normal sheep. DESIGN: Prospective study. SETTING: Tuscany. PARTICIPANTS: 100 eyes from 50 adult Massese female sheep were examined. The sheep included in the study were considered free of anterior ophthalmic abnormalities. PRIMARY AND SECONDARY OUTCOME MEASURES: Bacteria were identified by morphological assessment, Gram staining, biochemical tests. Identification of filamentous fungi was achieved at the genus level, and Aspergillus species were identified based on keys provided by other authors. Yeast colonies were highlighted, but not identified. RESULTS: Positive cultures were obtained from 100/100 eyes for bacteria, and from 86/100 eyes for fungi. A total of 14 types of bacteria and 5 types of fungi were isolated. Yeasts were isolated from 13/100 eyes. The most frequent fungal isolates were saprophytic fungi. CONCLUSIONS: Conjunctival bacterial and fungal flora of clinically normal eyes were reported in sheep. The positivity obtained for conjunctival bacteria was higher compared to findings in the literature by other authors in the same species (100 per cent v 40 per cent), while our results were in line with a recent work performed on mouflons (Ovis Musimon) with a 100 per cent positivity for bacterial conjunctival fornix. In our survey, Gram-positive species were prevalent, as reported by other authors in different species. Few data are available in the literature regarding conjunctival fungal flora in healthy small ruminants. The prevalence of conjunctival fungal flora in this study was higher than findings reported in mouflons (86 per cent v 45 per cent). Differences in fungal prevalence may be due to different methods of managing herds, though further studies are required to verify this hypothesis. The similarities in bacterial and fungal isolates between sheep and mouflons suggest a genera pattern of conjunctival colonisation by bacteria and fungi.

Seroprevalence, detection of DNA in blood and milk, and genotyping of Toxoplasma gondii in a goat population in Italy.

Toxoplasma gondii is the causative agent of a major zoonosis with cosmopolitan distribution and is known to be transmitted mainly by the ingestion of undercooked or raw animal products. Drinking unpasteurized goat's milk is a risk factor associated with human toxoplasmosis. However, very little is known about the excretion of DNA in goat milk. Aim of the present study was to determine the seroprevalence of T. gondii infection using a modified agglutination test (MAT), to detect T. gondii DNA by nested-PCR (n-PCR) in samples of blood and milk from seropositive goats, and to genotype DNA isolates using 11 molecular markers in 127 adult lactating goats from 6 farms in Italy. Positive MAT results were found in 60.6% of goats while 13% of blood and milk samples from seropositive goats were positive to n-PCR. A kappa coefficient of 1 indicated a perfect agreement between blood and milk n-PCR. Genetic characterization of isolates revealed the occurrence of genotype III (n = 7), genotype I (n = 1), and atypical genotypes with hints for genotype I (n = 2). Our results suggest that the risk of excretion of Toxoplasma tachyzoites might frequently occur in milk of seropositive goats testing positive to n-PCR on blood.

Serologic, molecular, and pathologic survey of Toxoplasma gondii infection in free-ranging red foxes (Vulpes vulpes) in central Italy.

We tested 191 sera of red foxes (Vulpes vulpes; 78 females and 113 males) for antibodies to Toxoplasma gondii using an indirect immunofluorescent test. Tissue samples of myocardium, lymph nodes, and brains from antibody-positive animals were tested for T. gondii DNA using specific PCR and processed for histopathology and immunohistochemistry (IHC) to detect T. gondii antigen. Of 192 (53.4%) antibody-positive animals, eight were positive by PCR on myocardium and one on brain. All DNA extracts were genotyped. Histopathology showed lesions characteristic of protozoan encephalitis; IHC did not show T. gondii antigen in examined tissues. The high antibody prevalence found in our study, which is the first in Italy, and the occurrence of polymorphic strains (combination of different type I and III alleles) divergent from typical T. gondii strains, suggests red foxes may be a sentinel of T. gondii in the environment.

A herbal antifungal formulation of Thymus serpillum, Origanum vulgare and Rosmarinus officinalis for treating ovine dermatophytosis due to Trichophyton mentagrophytes.

A number of herbal products with anti-inflammatory, antiseptic and antimycotic properties are available for dermatological usage. The successful treatment of 13 sheep affected by ringworm due to Trichophyton mentagrophytes with a mixture consisting of essential oils (EOs) of Thymus serpillum 2%, Origanum vulgare 5% and Rosmarinus officinalis 5% in sweet almond (Prunus dulcis) oil. The effectiveness of EOs and of the major components of the mixture (thymol, carvacrol, 1,8 cineole, α-pinene, p-cymene, γ-terpinene) against the fungal clinical isolate was evaluated by a microdilution test. Thirteen animals were topically administered with the mixture twice daily for 15 days. The other sheep were administered with a conventional treatment (seven animals) or left untreated (two animals). Minimum inhibitory concentration (MIC) values were 0.1% for T. serpillum, 0.5% for O. vulgare, 2.5% for I. verum and 5% for both R. officinalis and C. limon. Thymol and carvacrol showed MICs of 0.125% and 0.0625%. A clinical and aetiological cure was obtained at the end of each treatment regimen in only the treated animals. Specific antimycotic drugs licenced for food-producing sheep are not available within the European Community. The mixture tested here appeared to be a versatile tool for limiting fungal growth.

Toxoplasma gondii in waterfowl: the first detection of this parasite in Anas crecca and Anas clypeata from Italy.

Understanding the spread of Toxoplasma gondii in waterfowl is of interest for elucidating the potential involvement of these birds in maintaining the parasitic life cycle because birds are exposed to these parasites. Sera from 103 adult, free-range game birds, representing 13 different species living in the Italian wetlands, were examined using modified agglutination tests for antibodies specific to T. gondii . In seropositive birds, the brain and heart were homogenized and DNA was extracted to perform nested polymerase chain reaction (nPCR) and genotyping. Out of the 103 serum samples, 9 (8.7%) were scored as positive with titers ranging from 1/20 to 1/160. The seropositive animals corresponded to 4 of the species: Anas platyrhynchos (mallard; 2/17), Anas clypeata (northern shoveller; 2/11), Anas crecca (common teal; 3/41), and Gallinago gallinago (common snipe; 2/8). Parasite DNA was detected in 3 out of 9 brain samples, while the PCR results from the heart specimens were negative in all of the birds. The occurrence of non-clonal types of T. gondii was suggested in all cases. In conclusion, we describe the first documented detection of DNA of T. gondii in tissues from the northern shoveller and common teal, thereby extending the range of intermediate hosts for this parasite.

Prevalence of Toxoplasma gondii infection in Myocastor coypus in a protected Italian wetland.

BACKGROUND: Toxoplasma gondii is the causative agent for a major zoonosis with cosmopolitan distribution. Water has been implicated in outbreaks of toxoplasmosis in recent years. Coypus (Myocastor coypus), commonly nutria, are large semi-aquatic invasive rodents, naturalized throughout European countries, including most wetlands of Central Italy. The habitat of these animals is both terrestrial and aquatic, making them a species highly exposed to the parasite. FINDINGS: The occurrence of the infection was evaluated using a modified agglutination test (MAT) in 74 adult coypus from a naturalized population living in a wetland of Central Italy. Nested PCR (n-PCR) assay was carried out on some of them. Positive T. gondii MAT results were found in 44 animals (59·4%), 30 males (68·2%) and 14 females (31·8%). Antibody titers were ranging from 20 to 40960, while 12 out of 23 (52·2%), examined animals, 8 males (66·7%) and 4 females (33·3%), resulted positive to n-PCR. All n-PCR positive animals were seropositive, showing antibody titers ranging from 640 to 40960. CONCLUSIONS: Our results indicate that examined animals are heavily parasitized with Toxoplasma. This suggests that coypus could be a reservoir of this parasite, because they can be eaten both by scavenger animals and by humans, and that these animals would play a role in maintaining the cycle of T. gondii.