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Echinococcus equinus in a Horse: A Risk When Raw-Meat is Fed to Dogs?
J. Comp. Path. 2017, Vol. 156, 54e141
ESVP and ECVP Proceedings 2016
97
ECHINOCOCCUS EQUINUS IN A HORSE: A RISK WHEN RAW-MEAT IS FED TO DOGS? S. Merz *, N.A. Erickson *, J. Kr€ ucken y and A.D. Gruber* *Department of Veterinary Pathology and yInstitute of Parasitology and Tropical Veterinary Medicine, Freie Universit€at Berlin, Germany Introduction: Echinococcus equinus is a cestode with a two-host life cycle, with carnivores as definitive hosts, shedding eggs with the faeces, and equids as intermediate hosts, with hydatid cysts forming mainly in the liver and lung. Materials and Methods: A 17-year-old Connemara horse with an oromaxillary sinus fistula and chronic colitis was humanely destroyed with poor prognosis. The horse had a history of long-term corticosteroid therapy. Necropsy examination identified several well-demarcated, greyewhite cysts of up to 5 cm in diameter in the liver parenchyma, filled with clear, amber-coloured liquid containing particles of !1 mm in diameter (‘hydatid sand’). These cysts were further investigated by histopathology and polymerase chain reaction (PCR). Results: At necropsy examination, additional smaller cysts of up to 1 cm in diameter were visible in the liver and also in the lung. Histopathology revealed a trilayered appearance of the cysts with an outer connective tissue capsule followed by a PAS-positive, hyaline acellular layer and an inner germinal membrane. The cysts contained few free protoscolices indicating the presence of Echinococcus spp. PCR product sequencing revealed complete identity with E. equinus 12S rRNA and cytochromoxidase 1. Conclusion: The present case indicates that although E. equinus infections are still considered rare, they have emerged to our previously unaffected latitude. They pose a potential risk to dogs being fed with raw horse meat or liver in terms of the BARF (Biologically Appropriate Raw Foods) diet, which has gained increasing popularity. These dogs may potentially become reservoirs for E. equinus, a risk that should be considered.
DETECTION OF MAEDI-VISNA ANTIGEN IN MAMMARY GLAND AND MILK BY IMMUNOHISTOCHEMISTRY E. Gayo *, L. Polledo y, A. Balseiro z, M.J. Garcıa Iglesias *, C. P erez * and J.F. Garcıa Marın* *Pathological Anatomy Section, Animal Health Department, Veterinary School, University of Leon, Leon, yMicros Veterinaria, Leon and zSERIDA, Asturias, Spain Introduction: Macrophages are the main target cells of ovine maedivisna virus (MVV), although other cells such as epithelial cells have been suggested as possible targets of the disease. The aim of this study was to identify the target MVV cells in the mammary gland and their implication in MVV transmission through lactation in natural cases. Materials and methods: Mammary gland samples of 19 naturallyinfected sheep and 10 negative controls from intensive milkproducing flocks and milk samples from tank container, seropositive and seronegative animals were studied. Immunohistochemistry and immunocytochemistry against gp135 and p28 of MVV were carried out. Results: MVV infection was confirmed in the mammary gland and was always associated with lesions, even when these were minimal and focal. Macrophages were the only positive cells observed and were located in the interstitium (n 5 19), acinar lumen (n 5 2) and milk samples. No positivity was found in epithelial cells. Conclusions: The presence of MVV antigen in mammary gland and milk confirms the potential transmission of the disease through lactation. The presence of this virus was only associated with inflammatory cells, confirming the hypothesis of viral invasion via monocytes/macrophages, which would begin in focal locations and then spread to the rest of the tissue. No positivity was observed in epithelial cells, in contrast to the hypothesis of the possible MVV replication in these cells as has been described in vitro.
ANTIFUNGAL SUSCEPTIBILITY OF CANINE AND FELINE MALASSEZIA SPP. ISOLATES TO LACTOFERRICIN: PRELIMINARY IN-VITRO STUDY E. Biasibetti *, A. Corona y, F. Valenza *, F. Dosio z, N. Bruni x and M.T. Capucchio* *Department of Veterinary Sciences, University of Torino, Torino, y Ambulatorio Veterinario Associato, Torino, zDepartment of Drug Science and Technology, University of Torino, Torino and xIstituto Farmaceutico Candioli, Beinasco, Torino, Italy Introduction: Malassezia spp. is a fungus isolated from the skin and mucosae of dogs and cats, which may cause dermatitis and otitis. Therapies include the use of antibiotics, antifungals and glucocorticoids. Because of the antibiotic-resistance phenomenon, new alternative therapies are necessary. Bovine lactoferricin (Lfc) is a peptide derived from proteolytic cleavage of lactoferrin with proven antibacterial, antifungal and immunostimulatory activity. The aim of this study was to evaluate the antifungal susceptibility of Malassezia spp. to Lfc-Candioli Pharma (water solution 20%) using a microdilution method. Materials and Methods: Fifty strains of Malassezia spp. collected from 50 animals (five cats, 45 dogs) affected by dermatitis and/or otitis externa and classified based on clinical signs and/or skin biopsies were cultured on Sabouraud’s dextrose broth. Minimal inhibitory concentrations of Lfc were measured using the following concentrations: 13.3%, 10%, 6.7 %, 3.3% and 1.8%. Plates were incubated at 35 C and read 4 days after inoculation. To check the reproducibility of the procedure, all of the isolates were double tested and quality controls were performed. Results: All isolates were inhibited by Lfc with different minimum inhibitory concentration value. The product showed antifungal efficacy of 100% up to a dilution corresponding to 10% of Lfc. The first resistance was observed from 6.7% to the total resistance of 1.8%. Conclusions: These results suggest a potential antifungal efficacy of Lfc in vivo, even if in-vitro data should be considered with caution until standardized methods and correlation with clinical outcomes has been evaluated.
BESNOITIOSIS IN AN EUROPEAN DONKEY C. De Vries *, A. Santi y, K. Rickards z and G. Loesenbeck* *LABOKLIN GmbH & Co. KG, Bad Kissingen, Germany, yAgnese Santi, Prato, Italy and zThe Donkey Sanctuary Veterinary Department, Sidmouth, UK Introduction: A 3-year-old neutered male donkey was presented with multifocal, moderately firm, dull-white nodules, varying in size from 0.5e2.0 cm in diameter. The nodules were located on the penile shaft skin and showed depigmentation. Other locations on the body were not involved. Materials and Methods: Surgical excision of one large nodule and multiple smaller nodules was performed. All formalin-fixed samples were embedded in paraffin wax and routinely processed for histopathological examination. Results: Histopathological examination revealed the presence of multifocal, round to oval, protozoal cysts within the dermis, with a size of 150e500 mm. The mature cyst walls consisted of four distinct layers, including an outer, hyalinized, eosinophilic layer of collagen fibres, a thin homogeneous intermediate layer, a layer consisting of the cytoplasm of the host fibroblast with a compressed nucleus, and an inner layer that formed the parasitophorous vacuole. The vacuole was filled with numerous bradyzoites of 2 8 mm. There was a mild to moderate, superficial to mid-dermal infiltration of lymphocytes, plasma cells, macrophages and eosinophils, surrounding the cysts and blood vessels. The hair follicles were atrophic. The overlying epidermis showed mild acanthosis and orthokeratotic hyperkeratosis. Based on the histopathological findings, a diagnosis of Besnoitia spp. infection was made. The large nodule was diagnosed as a sarcoid (early stage). Conclusions: Besnoitiosis is an emerging disease in cattle in Europe. Few outbreaks have been reported in donkeys in the USA. To our knowledge, this is the first report in the literature of Besnoitia spp. infection in a European donkey.
ESVP and ECVP Proceedings 2016
97
ECHINOCOCCUS EQUINUS IN A HORSE: A RISK WHEN RAW-MEAT IS FED TO DOGS? S. Merz *, N.A. Erickson *, J. Kr€ ucken y and A.D. Gruber* *Department of Veterinary Pathology and yInstitute of Parasitology and Tropical Veterinary Medicine, Freie Universit€at Berlin, Germany Introduction: Echinococcus equinus is a cestode with a two-host life cycle, with carnivores as definitive hosts, shedding eggs with the faeces, and equids as intermediate hosts, with hydatid cysts forming mainly in the liver and lung. Materials and Methods: A 17-year-old Connemara horse with an oromaxillary sinus fistula and chronic colitis was humanely destroyed with poor prognosis. The horse had a history of long-term corticosteroid therapy. Necropsy examination identified several well-demarcated, greyewhite cysts of up to 5 cm in diameter in the liver parenchyma, filled with clear, amber-coloured liquid containing particles of !1 mm in diameter (‘hydatid sand’). These cysts were further investigated by histopathology and polymerase chain reaction (PCR). Results: At necropsy examination, additional smaller cysts of up to 1 cm in diameter were visible in the liver and also in the lung. Histopathology revealed a trilayered appearance of the cysts with an outer connective tissue capsule followed by a PAS-positive, hyaline acellular layer and an inner germinal membrane. The cysts contained few free protoscolices indicating the presence of Echinococcus spp. PCR product sequencing revealed complete identity with E. equinus 12S rRNA and cytochromoxidase 1. Conclusion: The present case indicates that although E. equinus infections are still considered rare, they have emerged to our previously unaffected latitude. They pose a potential risk to dogs being fed with raw horse meat or liver in terms of the BARF (Biologically Appropriate Raw Foods) diet, which has gained increasing popularity. These dogs may potentially become reservoirs for E. equinus, a risk that should be considered.
DETECTION OF MAEDI-VISNA ANTIGEN IN MAMMARY GLAND AND MILK BY IMMUNOHISTOCHEMISTRY E. Gayo *, L. Polledo y, A. Balseiro z, M.J. Garcıa Iglesias *, C. P erez * and J.F. Garcıa Marın* *Pathological Anatomy Section, Animal Health Department, Veterinary School, University of Leon, Leon, yMicros Veterinaria, Leon and zSERIDA, Asturias, Spain Introduction: Macrophages are the main target cells of ovine maedivisna virus (MVV), although other cells such as epithelial cells have been suggested as possible targets of the disease. The aim of this study was to identify the target MVV cells in the mammary gland and their implication in MVV transmission through lactation in natural cases. Materials and methods: Mammary gland samples of 19 naturallyinfected sheep and 10 negative controls from intensive milkproducing flocks and milk samples from tank container, seropositive and seronegative animals were studied. Immunohistochemistry and immunocytochemistry against gp135 and p28 of MVV were carried out. Results: MVV infection was confirmed in the mammary gland and was always associated with lesions, even when these were minimal and focal. Macrophages were the only positive cells observed and were located in the interstitium (n 5 19), acinar lumen (n 5 2) and milk samples. No positivity was found in epithelial cells. Conclusions: The presence of MVV antigen in mammary gland and milk confirms the potential transmission of the disease through lactation. The presence of this virus was only associated with inflammatory cells, confirming the hypothesis of viral invasion via monocytes/macrophages, which would begin in focal locations and then spread to the rest of the tissue. No positivity was observed in epithelial cells, in contrast to the hypothesis of the possible MVV replication in these cells as has been described in vitro.
ANTIFUNGAL SUSCEPTIBILITY OF CANINE AND FELINE MALASSEZIA SPP. ISOLATES TO LACTOFERRICIN: PRELIMINARY IN-VITRO STUDY E. Biasibetti *, A. Corona y, F. Valenza *, F. Dosio z, N. Bruni x and M.T. Capucchio* *Department of Veterinary Sciences, University of Torino, Torino, y Ambulatorio Veterinario Associato, Torino, zDepartment of Drug Science and Technology, University of Torino, Torino and xIstituto Farmaceutico Candioli, Beinasco, Torino, Italy Introduction: Malassezia spp. is a fungus isolated from the skin and mucosae of dogs and cats, which may cause dermatitis and otitis. Therapies include the use of antibiotics, antifungals and glucocorticoids. Because of the antibiotic-resistance phenomenon, new alternative therapies are necessary. Bovine lactoferricin (Lfc) is a peptide derived from proteolytic cleavage of lactoferrin with proven antibacterial, antifungal and immunostimulatory activity. The aim of this study was to evaluate the antifungal susceptibility of Malassezia spp. to Lfc-Candioli Pharma (water solution 20%) using a microdilution method. Materials and Methods: Fifty strains of Malassezia spp. collected from 50 animals (five cats, 45 dogs) affected by dermatitis and/or otitis externa and classified based on clinical signs and/or skin biopsies were cultured on Sabouraud’s dextrose broth. Minimal inhibitory concentrations of Lfc were measured using the following concentrations: 13.3%, 10%, 6.7 %, 3.3% and 1.8%. Plates were incubated at 35 C and read 4 days after inoculation. To check the reproducibility of the procedure, all of the isolates were double tested and quality controls were performed. Results: All isolates were inhibited by Lfc with different minimum inhibitory concentration value. The product showed antifungal efficacy of 100% up to a dilution corresponding to 10% of Lfc. The first resistance was observed from 6.7% to the total resistance of 1.8%. Conclusions: These results suggest a potential antifungal efficacy of Lfc in vivo, even if in-vitro data should be considered with caution until standardized methods and correlation with clinical outcomes has been evaluated.
BESNOITIOSIS IN AN EUROPEAN DONKEY C. De Vries *, A. Santi y, K. Rickards z and G. Loesenbeck* *LABOKLIN GmbH & Co. KG, Bad Kissingen, Germany, yAgnese Santi, Prato, Italy and zThe Donkey Sanctuary Veterinary Department, Sidmouth, UK Introduction: A 3-year-old neutered male donkey was presented with multifocal, moderately firm, dull-white nodules, varying in size from 0.5e2.0 cm in diameter. The nodules were located on the penile shaft skin and showed depigmentation. Other locations on the body were not involved. Materials and Methods: Surgical excision of one large nodule and multiple smaller nodules was performed. All formalin-fixed samples were embedded in paraffin wax and routinely processed for histopathological examination. Results: Histopathological examination revealed the presence of multifocal, round to oval, protozoal cysts within the dermis, with a size of 150e500 mm. The mature cyst walls consisted of four distinct layers, including an outer, hyalinized, eosinophilic layer of collagen fibres, a thin homogeneous intermediate layer, a layer consisting of the cytoplasm of the host fibroblast with a compressed nucleus, and an inner layer that formed the parasitophorous vacuole. The vacuole was filled with numerous bradyzoites of 2 8 mm. There was a mild to moderate, superficial to mid-dermal infiltration of lymphocytes, plasma cells, macrophages and eosinophils, surrounding the cysts and blood vessels. The hair follicles were atrophic. The overlying epidermis showed mild acanthosis and orthokeratotic hyperkeratosis. Based on the histopathological findings, a diagnosis of Besnoitia spp. infection was made. The large nodule was diagnosed as a sarcoid (early stage). Conclusions: Besnoitiosis is an emerging disease in cattle in Europe. Few outbreaks have been reported in donkeys in the USA. To our knowledge, this is the first report in the literature of Besnoitia spp. infection in a European donkey.