Dermatophytosis in animals: epidemiological, clinical and zoonotic aspects.

AIM: Dermatophytosis are the most frequent fungal infections of pets and livestock and play an important role in animal and human health due to their zoonotic potential. Another important aspect of these infections is linked to the economic consequences in farm animal and fur production systems. An overview of dermatophytosis in animals is described in this paper. Epidemiological, clinical and zoonotic aspects are addressed, considering individual species, both pets and farmed animals. METHODS: In particular, most recent investigations in the field of animal mycology, carried out in Central Italy, are reported, with particular reference to rabbit, ruminants, horse, dog, cat and some wild species. RESULTS: The information in this article show how dermatophytes infect a wide range of animals which may be in contact with human beings either directly or indirectly. Consequently they are frequently a source of infection for human beings who, vice versa, may sometimes become contagious for animals. CONCLUSION: Fungal pathogens derive their power to spread from contamination of the animal's habitat - whether the animal is a conventional pet or not, a farm animal or living in the wild. Thus if treatment of the animal or human patient is to achieve optimal efficacy, it needs to be associated with adequate environmental measures.

Diagnostic and clinical features of animal malasseziosis.

Malassezia yeasts infection represents a common clinical concern with a special regard to canine dermatology. The Authors review the main clinical features of malasseziosis in canine and feline medicine, summarizing predisposing factors and aetiopathogenesis of the yeasts' infection. A special reference was given to clinical and microscopical diagnosis.

Neospora caninum in Wild Waterfowl: Occurrence of Parasite DNA and Low Antibody Titers.

Thirty-five adult waterfowl (14 males and 21 females) representing various orders and species were sampled during the hunting season from 2015 to 2016. Antibodies to Neospora caninum were detected by IFAT on blood samples, while heart and brain were subjected to molecular analysis for the detection of Neospora caninum DNA. Twelve birds (34.3%) (6 Anas crecca , 3 Anas platyrhynchos , 2 Anas penelope , 1 Anas acuta ) showed antibodies versus N. caninum , while 10 animals out of 35 (4 A. crecca , 2 A. platyrhynchos , 2 A. penelope , 1 A. acuta , and 1 Vanellus vanellus ) scored positive for at least 1 DNA sample, with an overall prevalence of 28.6%. The present report shows for the first time the occurrence of antibodies and N. caninum DNA in waterfowl. The avian species investigated in the present report, which feed directly from the soil and/or water, would be able to ingest oocysts excreted by final canid hosts and could contribute to parasite transmission in the sylvatic cycle. To achieve a definitive result about the role of these avian species in the epidemiology of this protozoan, the presence of viable parasites should be demonstrated by bioassay and/or culture, as well as histological evidence of N. caninum cysts in avian tissues.

Detection of Chlamydia pneumoniae in atherosclerotic coronary arteries.

Chlamydia pneumoniae has recently been associated with the development of coronary heart diseases by sero-epidemiological studies and by direct detection of the organism in atherosclerotic tissues. The aim of our study was to employ a semi-nested PCR approach to investigate the presence of C. pneumoniae in both normal and atherosclerotic coronary arteries of humans obtained at autopsy. Moreover, we have evaluated the role of infection with C. pneumoniae in relation to the extent of coronary atherosclerosis. One hundred and eighty coronary artery specimens were collected at autopsy from 60 consecutive subjects (three arterial segments from each subject). Atherosclerosis in each arterial segment was graded histologically by the Stary classification. Thirty normal coronary arteries were also taken at autopsy as control. PCR results evidenced the presence of C. pneumoniae DNA in atherosclerotic coronary arteries in 19 (31.7%) of 60 subjects examined, while none of the 30 subjects with non-atherosclerotic tissues was positive (p=0.001). Moreover, of the 180 atherosclerotic specimens examined, C. pneumoniae DNA was detected in 3.4% (2/59) of mild atherosclerotic lesions, and in 14.0% (17/121) of advanced atherosclerotic lesions (p=0.05). Our results demonstrate that the presence of C. pneumoniae DNA may be associated with the severity of coronary atherosclerosis.

Oral focal mucinosis of the tongue.

Oral focal mucinosis (OFM) is an uncommon clinicopathological entity which is considered to be the oral counterpart of cutaneous focal mucinosis and cutaneous myxoid cyst. It is comprised of a clinically elevated mass with a histological feature of localised areas of myxomatous connective tissue. The present study adds a rare case of OFM of the tongue to the literature, and we present a review of the most characteristic oral myxomatous lesions.

Benign tumors of heterotopic tissue in the thyroid gland: a report of two cases of lipomatous lesions.

A report of two cases, concerning heterotopic nests of fat cells in the thyroid gland, is presented here together with a review of lipomatous lesions in the literature. Both cases involved patients who presented with goiter; one had Grave's disease and the other had adenomatous hyperplasia. The fat cells were principally located in the subcapsular areas and scattered among the follicles. The distribution of the immunohistochemical staining, and the morphologic characteristics of the adipose tissue, suggested a probable origin of the fat cells from inclusion nests during embryogenesis of the thyroid gland.

Expression of bcl-2, c-erbB-2, p53, and p21 (waf1-cip1) protein in thyroid carcinomas.

The study was carried out on 53 patients who had thyroid cancer with various degree of differentiation. We studied the expression of bcl-2, a-erbB-2, p53, and p21 ras protein. The protein encoded by bCL-2 proto-oncogene is implicated in the prolongation of cell survival by blocking programmed cell death, i.e. apoptosis. The role of p53 and bcl-2 genes in the regulation of apoptosis has important implications in oncogenesis. Wild-type p53 is thought to promote apoptosis, whilst mutant p53 has a similar effect on apoptosis as bcl-2 that is inhibition of programmed cell kinase activity. C-erb-2 protein overexpression is currently being evaluated as a potential risk factor in breast cancer patients? The ras gene family codes for a 21 kD protein (p21), which binds guanine nucleotides and possesses GTPase activity. Through this mechanism, the ras p21 protein participates in the control of cell proliferation, possibly as a signal transducer from cell surface receptors to the nucleus. Activation of ras genes has been implicated in neoplastic transformation of cells. The aim of our study is to evaluate the expression of these markers in thyroid carcinomas. All immunohistochemical study was performed in paraffin-embedded tissues pathology specimen. Any well differentiated tumor in our study was positive for bcl-2 protein. C-erb-2 immunostaining was present in tumor samples in 60% of cases. In most cases, specific membrane staining as well as a weak cytoplasmic positivity of tumor cells were seen. Immunoreactivity for p53 was positive only in 10% of cases. By immunostaining, p21 protein was expressed in 55% of the 53 tumors tested, with different degree of expression. Only some poorly differentiated tumours were positive for bcl-2, furthermore all markers tested were strongly positive in these tumours. In conclusion, our results indicate that bcl-2, c-erbB-2, p53, and p21 ras protein are differently expressed in thyroid carcinomas in relation to the degree of aggressiveness and differentiation.

Feline cutaneous phaeohyphomycosis due to Cladophyalophora bantiana.

A case of feline cutaneous phaeohyphomycosis due to Cladophyalophora bantiana is described. The cat was presented with breathing difficulty and a swollen, ulcerated nodule on the dorsal nose and left nostril. Histological examination of the nodule revealed a cystic granulomatous dermatitis characterised by neutrophils, macrophages and giant cells. Pigmented, yeast-like fungus cells and hyphal elements were easily identified in haematoxylin-eosin stained tissue sections. Cladophyalophora bantiana was isolated from a tissue specimen. This organism, primarily known to cause cerebral infection in humans and cats, only rarely causes cutaneous infection. Despite anti-fungal chemotherapy two relapses occurred. The cat was feline immunodeficiency virus- and feline leukemia virus-negative and even if the owner was unaware of trauma, the hypothesis of wound contamination is the most likely.

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.

[Prognostic markers of the epithelial tumors of the large intestine]. / Indicatori prognostici della patologia tumorale di tipo epiteliale del grosso intestino.

The surgical treatment of large bowel malignant tumors is now the most important therapeutic approach. The aim of our study was to evaluate a possible prognostic implication of ploidy and NM23 expression. The study includes 120 non selected patients who underwent surgery for colorectal cancer. The analysis of ploidy was obtained with cytometric test. The cases with only one gaff G0/G1 were considered as diploid. Patteras with more than one aneuploidic population were classified as multiploidic or poliploidic. While NM23 expression was evaluated with a double blind retrospective study by two separate equipe of authors in different centres. NM23 positivity degree was classified in 3 classes: absent or weak if < 10%; moderate if 10-50%; strong if > 50%. Forty-two patients (35%) were classified as diploidic; 30 patients (25%) as aneuploidic; 40 patients (33.2%) as multiploidic; in 8 cases (6.7%) the material was inadequate. Among the two groups who evaluated NM23 expression the group I a positivity: absent or weak in 32 cases (26.7%); moderate in 62 cases (51.7%); strong in 26 cases (21.6%); while the group II shows positivity: absent or weak in 30 cases (25%); moderate in 52 cases (43.3%); strong in 38 cases (31.7%). We cannot consider ploidy and the NM23 research as independent prognostic factor.

A preliminary study on the quality and safety of milk in donkeys positive for Toxoplasma gondii.

Toxoplasmosis is one of the five parasitic diseases considered as a priority for public health action. The consumption of raw milk products represents a possible risk, in particular for certain categories of people. The aim of this study was to evaluate the possible effects of Toxoplasma gondii on milk yield and quality in sero-positive animals with parasitemia. Eighteen healthy lactating Amiata jennies, between 90 and 180 days were included in the study. Four donkeys scored positive for immunofluorescent antibody test (IFAT), and each IFAT positive donkey presented parasitic DNA both in the blood and milk. No significant differences were found between milk yield in PCR-positive donkeys compared with the negative cases, however the former tended to have a greater production. Milk quality in the positive donkeys showed a significantly lower percentage of casein (0.72% v. 0.81%) and ash (0.32% v. 0.37%). Positive cases had a highly significant larger average diameter of globules (2.35 µm) and fewer globules/ml (2.39 × 10(8)). Somatic cell and bacterial counts were normal and in agreement with the literature. Toxoplasma gondii did not seem to present clinical forms in lactating jennies. Further in vivo studies are needed to further assess the risk of T. gondii transmission through donkey milk, together with the impact of different stages of infection on milk quality.

Clinical and mycological evaluation of an herbal antifungal formulation in canine Malassezia dermatitis.

OBJECTIVE: Malassezia pachydermatis is a common cause of more widespread dermatitis in dogs (CMD). Recurrences are common, and this disorder can be very troubling for both dogs and for the pet owner. MATERIAL AND METHODS: The treatment of 20 dogs affected by dermatitis due to M. pachydermatis, with Malacalm(®), a commercially available mixture consisting of essential oils (Citrus aurantium 1%, Lavandula officinalis 1%, Origanum vulgare 0.5%, Origanum majorana 0.5%, Mentha piperita 0.5% and Helichrysum italicum var. italicum 0.5%, in sweet almond oil and coconut oil) is reported. The effectiveness of the whole mixture, of component essential oils and of their more represented compounds against clinical isolates was evaluated by a microdilution test. Twenty animals were topically administered the mixture twice daily for 1 month. Ten animals were treated with a conventional therapy based on ketoconazole 10mg/kg/day and chlorhexidine 2% twice a week for 3 weeks. At the end of both treatments animals significantly improved their clinical status. Adverse effects were never noticed. Follow-up visit performed on day 180th allowed to observe a recurrence of clinical signs in all the subjects treated conventionally, while not significant clinical changes were referred in dogs treated with Malacalm(®). The overall MIC value of Malacalm(®) was 0.3%. O. vulgare showed the lowest minimum inhibitory concentrations (MIC), being active at 0.8%, followed by M. piperita (1%), O. majorana (1.3%), C. aurantium (2%) and L. officinalis (4%) while H. italicum did not yield any antimycotic effect up to 10%. Active major compounds were thymol, carvacrol, p-cymene, 1,8-cineol, limonene and menthol. CONCLUSION: The phytotherapic treatment achieved a good clinical outcome, and no recurrence of skin disorders on day 180th was recorded. This herbal remedium appeared to be a safe tool for limiting recurrences of CMD.

Canine and feline dermatophytosis due to Microsporum gypseum: a retrospective study of clinical data and therapy outcome with griseofulvin.

OBJECTIVE: Microsporum gypseum is a common inhabitant of the soil, occasionally responsible for human and animal ringworm. Few reports describe the treatment of dermatologic diseases due to M. gypseum. The objective of this study was to evaluate retrospectively cases of M. gypseum infection in dogs and cats. MATERIAL AND METHODS: The occurrence of infection by this dermatophyte was retrospectively evaluated in dermatological specimens from 15,684 dogs and cats dermatologically diseased from Italy. Clinical outcome after treatment with griseofulvin combined with topical enilconazole was evaluated in 41 dogs and, out of label, 10 cats. Furthermore, in vitro susceptibility to griseofulvin and enilconazole was evaluated on 31 clinical isolates of M. gypseum. RESULTS: One hundred and eighty-five specimens out of 15,684 (1.1%) scored positive for M. gypseum. The treatment failed to achieve both mycological and clinical cure in 16 dogs (39%) and four cats (40%), as well as fungal isolates demonstrated a very poor in vitro sensitivity when tested versus griseofulvin: the MIC value was 150 µg/mL. The ED50 value was calculated at 66 µg/mL. CONCLUSION: Blind treatments with griseofulvin in ringworm due to M. gypseum should be avoided.

In vitro and in vivo antifungal activity of some essential oils against feline isolates of Microsporum canis.

The treatment of dermatophytoses due to Microsporum canis is cumbersome and relapses can occur. Volatile essential oils (EOs) obtained from plants would seem to represent suitable tools to contrast mycoses both in human and animals. The anti-M. canis activity of some EOs chemically characterized was evaluated both in vitro and in vivo. Eleven feline isolates of M. canis were tested by microdilution against EOs extracted from Thymus serpillum, Origanum vulgare, Rosmarinus officinalis, Illicium verum and Citrus limon. A mixture composed by 5% O. vulgare, 5% R. officinalis and 2% T. serpillum, in sweet almond oil was administered to seven infected, symptomatic cats. T. serpillum and O. vulgare showed the lowest MICs, followed by I. verum, R. officinalis and C. limon. The assay performed on mixture showed that antimycotic activity of each component was enhanced. Four out of seven treated cats recovered both clinically and culturally. T. serpillum and O. vulgare EOs showed a strong antifungal activity. Preliminary data suggest a possible application in managing feline microsporiasis. Considering the potential zoonotic impact of this infection, the use of alternative antimycotic compounds would be of aid to limit the risk of environmental spreading of arthrospores.

Presence and distribution of fungi and bacteria in the reproductive tract of healthy stallions.

A saprophytic bacterial flora is present on the penis and the distal part of the urethra of stallions. Little is known about the fungal flora of their reproductive tract. As micro organisms play an important role in mares fertility, the aim of the study was to describe the distribution of fungi and bacteria in the normal genital apparatus of stallions. The microbic flora of the reproductive tract of 11 healthy, fertile stallions was evaluated, collecting samples from 5 different locations: urethral fossa, penis/internal lamina of the prepuce, urethra pre- and post-ejaculation, and semen. For fungal examination samples were taken on 3 different occasions (N = 165), while for bacteriologic examination samples were taken on one occasion only (N = 55). There was a statistical difference in the presence of filamentous fungi between urethral fossa or penis/prepuce (45.4%) and urethra pre- or postejaculation or semen (15.1%, 6.0%, and 0.0%, respectively). Yeasts were isolated in 9.1% of the samples, never in semen. The most represented mycelial fungi were Penicillium spp., Aspergillus spp., Scopulariopsis spp., Trichosporon spp. and Mucoracee. The proportion of samples showing a total bacterial count ≥ 10,000 colony forming units (CFU) was higher for urethral fossa than for urethra pre- or postejaculation or for semen. Some bacterial growth was always observed in all locations, including the ejaculate. Differences between sampling locations were observed also for Staphylococci, both coagulase positive and negative. Salmonella enterica Abortus equi and sulphite reducing clostridia and other pathogens (including Klebsiella spp. and Pseudomonas spp.) were never isolated. Escherichia coli and coliforms always showed a low or absent flora. These data add information to the literature.

Isolation and characterization of Malassezia spp. in healthy swine of different breeds.

Malassezia spp. genus is represented by several lipophilic yeasts, normally present on the skin of many warm-blooded vertebrates, including man. Swine are one of the less investigated animal species. The aim of the present work was to study the occurrence of Malassezia spp. in the external ear canal of 408 healthy swine of different breeds, under different breeding conditions. For this purpose N. 185 free-ranging wild boars, N. 107 large size pigs and 116 Cinta Senese breed were selected. Animals were of both genders, with age ranging from 8 months to 4 years. The subjects were culturally and molecularly checked for Malassezia spp. Ninety-two out of 408 animals scored positive for Malassezia yeasts (22.5%). Malassezia pachydermatis, Malassezia sympodialis and Malassezia furfur were recognized. M. pachydermatis was the sole species isolated from wild boars (12.9%), Cinta Senese (20.7%) and juvenile large size pigs (13.6%); 88% of large size breeds adult subjects scored positive for M. sympodialis (63.6%) and M. furfur (22.7%), respectively. The study focus on scarcely investigated epidemiological aspects of Malassezia spp. in this animal species.

High infection rate of Trichophyton verrucosum in calves from Central Italy.

Recent epidemiological features of the zoonotic dermatophyte Trichophyton verrucosum were investigated in 294 calves from 20 farms located in Central Italy. By the combination of microscopic examinations and fungal cultures, the total prevalence rate of T. verrucosum infection was found to be high, with 87.7% of samples yielding positive results and 100% of farms being infected. Farm to farm prevalence of infection varied from 25% to 100% of the examined calves. Prevalence rates were higher in females than males (91.6% versus 84%), in >3-< or =6 than >6-< or =9 month old calves (89.8% versus 85.7%), in purebred than in crossbred (88.5% versus 81.8%), and in symptomatic than asymptomatic ones (95.7% versus 80.4%). T. verrucosum was as prevalent in semi-intensive as in intensive breeding system (88.2% versus 87.6%) but more prevalent in calves raised for meat rather than milk production (91.1% versus 84.9%), and reared under conventional than organic production management system (89.3% versus 82.6%). Prevalence rates were also higher when the cattle present in the farm were of mixed origin, rather than when the whole stock was purchased or was born in the farm (92% versus 88.2% and 85.7%), when other cattle in the farm were showing skin signs (90% versus 84.2%), and at least one case of human ringworm was recorded in the farm (90.9% versus 86.1%). No statistically significant association was found. Therefore, T. verrucosum infection may be more widespread in calves from Central Italy than it was previously thought to be. Calves of any sex, breed, age, and of any of the farm condition considered, are highly exposed to the risk of infection. As calves may play a considerable role as reservoirs of the infection, our results highlight the need to determine the incidence and the importance of T. verrucosum ringworm in personnel involved with cattle care in Italy and elsewhere.

Anatomopathological aspects of avian aspergillosis.

Aspergillosis is a fungal disease caused by fungi of the genus Aspergillus, in particular A. fumigatus and A. flavus. This paper focuses on anatomopathological aspects resulting from a chronic infection from Aspergillus spp in the chicken (Gallus domesticus), in the herring gull (Larus cachinnans micaelli) and in the red-legged partridge (Alectoris rufa rufa). Microscopically, we observed some histological lesions that are related to the two typical forms of Aspergillosis: a deep nodular form, typical of organs with a non-aerated parenchyma, and a non-encapsulated superficial diffuse form typical of the serosae and the lung. The observed forms of aspergillosis have been found in animals raised in poor hygienic environmental conditions or malnourished animals (chicken); in wild birds from wildlife recovery centres (herring gull), which underwent some forms of stress, such as traumas, detention, starvation, extended antibiotic treatments; in game birds (red-legged partridge) used for restocking natural areas that had been negatively affected by such stressors as captivity in aviaries, containment and transport in cages, release in unsuitable environments and malnutrition. The observed anatomopathological and istopathological aspects can therefore be regarded as the outcome of a number of factors that have reduced the typical resistance of the species and impaired the efficiency of their immune systems.

Antifungal activity of tea tree oil from Melaleuca alternifolia against Trichophyton equinum: an in vivo assay.

Dermatophytes are a group of keratinophilic and keratinolytic molds, some of which are responsible for ringworm. Among them Trichophyton equinum, which mostly infects equids, can cause extensive outbreaks in stud farms. The conventional treatment of equine trichophytosis is topic, based upon medicated shampoos to reduce the spread of infection among the animals. Nevertheless the popularity of phytotherapy is at an all-time peak, and the interest for natural alternatives or complements to conventional drug therapy is challenging both in human and veterinary field. Among herbal remedia Tea Tree Oil (TTO) shows a wide range of antimicrobial activities. A randomized open clinical trial was carried out on 60 thoroughbred breeding horses affected by equine ringworm. The animals were randomly divided into 2 groups of 30 subjects. Diagnostic criteria were the presence of clinical signs and positive T. equinum culture. Specificity control using TTO mixture in 5 not dermatophyte affected animals was achieved also. The antimycotic activity against T. equinum of a mixture containing 25% TTO in sweet almond oil, was evaluated in vivo treating 30 subjects, the others were administered enilconazole 2% solution. The animals of both groups were topically treated twice a day for 15 days with a 25% mixture of TTO diluted in sweet almond oil and every 3 days, four times with enilconazole rinses, respectively. The clinical and mycological outcome were evaluated at day 30 from the start of the treatments. Data analysis was performed by chi square test. All the treated animals showed complete clinical and aetiological healing. Part of control subjects also, showed an improvement and none of them exacerbate the lesions. This therapeutic protocol appears to be effective and versatile, being applicable immediately after physical examination, prior to have the laboratory response. It could be an alternative for practitioners interested in herbal medicines, contributing to fulfill the gap existing between in vitro and clinical studies.