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.

Sarcoptic mange and other ectoparasitic infections in a red fox (Vulpes vulpes) population from central Italy.

Fifty red foxes (Vulpes vulpes) from the district of Pisa (central Italy) were examined for ectoparasites. Sarcoptic mange was diagnosed on the presence of clearly visible skin lesions with confirmatory demonstration of Sarcoptes scabiei at parasitological and histopathological analysis. Ticks and fleas were collected directly from the carcases during post mortem examination, fixed and identified by morphological examination. For the detection of ear Malassezia and mite infections, cytological and parasitological examinations of ear wax samples were performed. All data were statistically analysed using a χ2 test with the Yates correction. An overall prevalence of 84% for ectoparasitic infections was found in examined subjects. In regard to isolated ectoparasites, 38%, 8%, 82%, 6% and 8% of foxes resulted positive for S. scabiei, Otodectes cynotis, Malassezia spp., fleas (Archaeopsylla erinacei, Pulex irritans, Ctenocephalides canis) and ticks (Ixodes ricinus and Rhipicephalus sanguineus), respectively. Malassezia ear infection was significantly more prevalent in animals older than 1 year (P < 0.01). Prevalence (38%), severity of lesions and poor body conditions observed in most Sarcoptes-infected animals indicate that sarcoptic mange should be considered the most important ectoparasitic infection of red foxes in the examined area.

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.

Retrospective study of 14 cases of canine arthritis secondary to Leishmania infection.

OBJECTIVE: To describe the clinical appearance, laboratory findings and response to treatment of dogs with inflammatory joint disease associated with Leishmania infection. METHODS: Retrospective analysis of case records of dogs with serologically confirmed leishmaniasis and concurrent inflammatory joint disease presented between 2005 and 2011. RESULTS: In total, 14 cases met the inclusion criteria. Of these, five (36%) dogs were presented with monoarthritis, five (36%) with oligoarthritis and four (28%) with polyarthritis. The most frequently affected joint was the carpus. Both erosive and non-erosive disease was identified on radiographic examination. All dogs had an inflammatory synovial fluid with a high white cell count and a preponderance of neutrophils, and in eight (57%) cases Leishmania amastigotes were found in the synovial fluid smears. Dogs were treated with 50 mg/kg N-methylglucamine antimoniate twice a day for 1 month and 10 mg/kg allopurinol twice a day for 6 to 9 months combined with prednisolone in five cases. At the 6-month follow-up, eight (57%) dogs showed improvement in general and orthopaedic signs and four (28%) dogs were stable. CLINICAL SIGNIFICANCE: Leishmaniasis should be considered a differential diagnosis in dogs with inflammatory arthritis in endemic areas.

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.

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.

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.

Coprological survey in pet reptiles in Italy.

Faecal samples were collected from 324 pet reptiles showing no clinical signs, including 28 saurian species (n=192), three ophidian species (n=74) and three chelonian species (n=58). Samples were examined for the presence of intestinal parasites by direct smear and faecal flotation, while direct immunofluorescence assays were used to reveal the presence of Cryptosporidium oocysts and Giardia cysts. Overall, 57.4 per cent of the reptiles were harbouring intestinal parasites. These included oxyurids (16 per cent), coccidia (12.3 per cent), flagellates (9.3 per cent), strongyles (6.8 per cent), coccidia plus oxyurids (4.9 per cent), coccidia plus flagellates (1.8 per cent), coccidia plus strongyles (1.8 per cent), oxyurids plus strongyles (1.2 per cent), oxyurids plus flagellates (1.2 per cent), Cryptosporidium species (1.2 per cent) and strongyles plus flagellates (0.6 per cent). Intestinal parasites were more prevalent in saurians than in ophidians and chelonians, in insectivores than in carnivores, omnivores and herbivores, and in wild-caught than in captive-born reptiles. A highly significant difference was observed for saurians versus chelonians (odds ratio [OR]=2.20, 95 per cent confidence interval [CI] 1.21 to 3.99), insectivores versus herbivores (OR=2.38, 95 per cent CI 1.26 to 4.49) and in wild-caught versus captive-born pet reptiles (OR=2.36, 95 per cent CI 1.27 to 4.40).

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.

Effectiveness of praziquantel for treatment of peritoneal larval cestodiasis in dogs: a case report.

Peritoneal larval cestodiasis (PLC) was incidentally identified in an adult female mixed-breed dog by explorative celiotomy done in response to abnormal blood values found during a routine work-up in preparation for an ovariohysterectomy. Ten days after ovariohysterectomy, treatment with fenbendazole began (50mg/kg, per os, every 12h, for 21 days). Two weeks after the end of treatment, samples of peritoneal fluid were obtained by paracentesis and examined. Motile peritoneal tetrathyridia were still evident. Fenbendazole was discontinued. After 10 days of withdrawal from fenbendazole, the dog was treated with a subcutaneous administration of injectable praziquantel (5mg/kg). The administration was repeated after a 15 days interval. Two weeks after the second administration, samples of peritoneal fluid were obtained again by paracentesis. Motile peritoneal tetrathyridia were not detected. Fourteen months after the last administration of praziquantel, the dog remained still free of peritoneal tetrathyridia as determined by abdominal ecography. Therefore, praziquantel was effective to eliminate peritoneal tetrathyridia definitely. Practitioners should be aware of PLC in order to early recognize this condition in case of incidental finding and implement adequate therapy as soon as possible.

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.

Occurrence and cyst burden of Giardia duodenalis in dog faecal deposits from urban green areas: Implications for environmental contamination and related risks.

To investigate the prevalence and cyst burden of Giardia duodenalis as well as the genetic identity and possible zoonotic potential of isolates in canine faeces left on soil, a total of 143 freshly passed faecal specimens were collected in four different green urban areas and examined by Real-time PCR. Overall, 30.8% of faeces from different dogs that defecated in urban areas were found to harbour Giardia cysts, with single area positivity proportions ranging from 4.2% to 47.8%; significant (P<0.05) and highly significant (P<0.01) differences were found in the prevalence among different areas investigated. The number of cysts ranged from 2 to 1428 per gram of faeces. Results emphasize that a high risk of dog-to-dog Giardia transmission may occur in public areas where high quantities of faeces from infected dogs are daily left to cumulate on the ground. Results suggest there is a low risk for zoonotic transmission of Giardia: only 0.6% of canine faeces contaminating urban areas were carrying subgenotype A1, which is considered as the most potentially transmissible to humans.

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.

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.

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.

Isolation of Scopulariopsis spp. fungi from Psoroptes cuniculi body surface and evaluation of their entomopathogenic role.

In this study, some in vitro trials were carried out to evaluate the association between Scopulariopsis spp. fungi with Psoroptes cuniculi (Acari: Psoroptidae) and their potential pathogenicity to this mite species. After cultivation on Sabouraud dextrose agar with chloroamphenicol at 26 degrees C for 20 days and macro- and microscopical examinations, from P. cuniculi mites taken from some infested rabbits fungi belonging to the genus Scopulariopsis were isolated. Investigations were carried out to evaluate in vitro the potential pathogenic role of Scopulariopsis to P. cuniculi; to this aim, Scopulariopsis brevicaulis isolated from a cat was tested. In several culture media, the dose-dependent P. cuniculi mortality with different concentrations of S. brevicaulis and the ability of S. brevicaulis to penetrate inside the body of infected mites were evaluated. Results obtained demonstrated that, in the rabbit, Scopulariopsis fungi can be associated with P. cuniculi, and that S. brevicaulis can be an entomopathogen for P. cuniculi in a dose-dependent manner. A more rapid mortality of the mites and a quicker S. brevicaulis growth in plates containing rabbit serum with respect to all other media used were also found. S. brevicaulis demonstrated the ability to invade the body of P. cuniculi.