Establishment of clinical criteria for the diagnosis of adverse food reactions in dogs with atopic dermatitis.

BACKGROUND: Diagnosis of canine adverse food reactions (AFRs) is based on vague criteria, such as '>50% improvement' during elimination diet trial (EDT) followed by 'deterioration' during provocation test (PT). OBJECTIVE: The objective of the study was to use predefined criteria to evaluate response during EDT [i.e., Owner Global Assessment of Treatment Efficacy (OGATE) = good-to-excellent] and relapse during PT [i.e., Owner Global Assessment of Challenge Deterioration (OGACD) = moderate-to-severe and/or >100% increase of lesional (Canine Atopic Dermatitis Extent and Severity Index, 4th iteration, CADESI-04) and/or of pruritus (pruritus Visual Analog Scale, PVAS) scores]. ANIMALS: Twenty-nine dogs with atopic dermatitis. MATERIALS AND METHODS: An extensively hydrolysed diet was fed to all dogs followed, in seven of 11 nonresponders, by a second home-made novel-protein EDT. Dogs responding to either EDT were challenged with their previous diet. RESULTS: Thirteen (44.8%) dogs were diagnosed with AFRs: at the end of EDT, their OGATE was good (9 of 13; 69.2%) or excellent (four of 13; 30.8%), and both CADESI-04 (46.7%) and PVAS (71.1%) had decreased significantly; at the end of PT, OGACD was moderate or severe in 12 of 13 (92.3%) dogs, and both CADESI-04 (127.9%) and PVAS (181.8%) had increased significantly. Of the 16 dogs without AFRs, 6 (37.5%) responded to the commercial (n = 5) or home-made (n = 1) diet [OGATE = good (three of six) or excellent (three of six)], with significant concurrent reduction of CADESI-04 and nonsignificant reduction of PVAS, yet they did not relapse during PT. CONCLUSIONS AND CLINICAL RELEVANCE: The proposed (predefined) criteria for the evaluation of response during EDT and deterioration during PT seem reliable and are easily applicable in clinical practice and research.

Specificity of a pancreatic lipase point-of-care test and agreement with pancreatic lipase immunoreactivity in cats without clinical evidence of pancreatitis.

OBJECTIVES: The aim of this study was to determine the specificity of a rapid point-of-care test for the estimation of feline pancreatic lipase (SNAP fPL) in healthy and sick cats without clinical evidence of pancreatitis. A second objective was to evaluate the agreement between SNAP fPL and serum pancreatic lipase immunoreactivity (fPLI), as measured by Spec fPL. METHODS: A total of 150 cats were prospectively enrolled into this study. Of them, 82 cats were healthy while 68 cats had various diseases but no clinical signs (eg, anorexia, depression, vomiting) raising a suspicion of pancreatitis. RESULTS: SNAP fPL was normal in 133/150 cats (specificity 89%) without obvious clinical pancreatitis. SNAP fPL was normal in 74/82 healthy cats (specificity 90%) and in 59/68 cats that were sick but without typical signs of pancreatitis (specificity 87%). The agreement between SNAP fPL and Spec fPL was substantial (k = 0.64) in healthy cats and almost perfect (k = 0.93) in sick cats. The overall agreement between SNAP fPL and Spec fPL was almost perfect (k = 0.81). CONCLUSIONS AND RELEVANCE: The specificity of SNAP fPL in this group of cats was high. There was a substantial and almost perfect agreement between the SNAP fPL and Spec fPL in healthy cats and sick cats without suspected pancreatitis, respectively. In the small percentage of cats with abnormal SNAP fPL and/or Spec fPL results, the possibility of subclinical pancreatitis cannot be excluded.

Seroprevalence of and Risk Factors for Toxoplasma gondii Infection in Cats from Greece.

Toxoplasmosis is one of the most important protozoan diseases with a global impact on the health of domestic cats and with zoonotic significance. The aims of this study were to determine the prevalence of seropositivity for Toxoplasma gondii in different populations of cats in Greece and to assess risk factors for seropositivity. A total of 457 cats were prospectively enrolled, and a commercially available indirect immunofluorescence antibody testing (IFAT) kit was used for the detection of anti-T. gondii immunoglobulin G (IgG) in serum. Overall, 95 (20.8%) of the 457 cats were seropositive for T. gondii. Based on multivariate analysis, factors associated with seropositivity included older age [Odds ratio (OR), 1.33; p < 0.001]; a history of cat-fight trauma (OR, 3.88; p = 0.004); and lack of vaccination against calicivirus, herpesvirus-1, panleukopenia, and rabies (OR, 10; p = 0.002). This study shows a high prevalence of seropositivity for T. gondii in cats in Greece. This implies that toxoplasmosis is still a major public health concern and that optimal strategies for the prevention of infection with T. gondii in cats should be established.

Investigation of comorbidities in dogs with leishmaniosis due to Leishmania infantum.

In endemic areas, dogs with leishmaniosis due to Leishmania infantum frequently have comorbidities, including mostly neoplastic, infectious, and parasitic diseases. The aim of this study was to compare the prevalence of comorbidities among dogs that are not infected by L. infantum, dogs that are infected but do not present leishmaniosis, and dogs with leishmaniosis, and to examine if certain comorbidities are independent risk factors for the infection by L. infantum and/or for the development of canine leishmaniosis (CanL). A total of 111 dogs, older than 1-year and non-vaccinated against CanL, were allocated into three groups: group A (n = 18) included dogs that were not infected by L. infantum, group B (n = 52) included dogs that were infected by L. infantum but did not present CanL, and group C (n = 41) included dogs with CanL. Signalment and historical data were obtained using a structured questionnaire. Laboratory examinations included complete blood count, serum biochemistry, urinalysis, fecal parasitology, modified Knott's test, microscopic examination of capillary blood, buffy coat, lymph node, bone marrow and conjunctival smears, qualitative serology for Dirofilaria immitis, Anaplasma phagocytophilum/A. platys, Borrelia burgdorferi and E. canis, IFAT for L. infantum, ELISA for Babesia spp. and Neospora caninum, and real-time PCR for L. infantum in bone marrow, skin biopsies and conjunctival swabs. A variety of comorbidities were found in all three groups. No independent risk factors for infection by L. infantum were found. On the contrary, among dogs infected by L. infantum, being a mongrel [odds ratio (OR): 11.2], not receiving prevention for dirofilariosis (OR: 26.5) and being seropositive to N. caninum (OR: 17.1) or to Babesia spp. (OR: 37.6), were independent risk factors for presenting CanL. Although no comorbidities influence the probability of canine infection by L. infantum, certain comorbidities may be precipitating factors for the transition from the subclinical infection by L. infantum to the overt CanL.

Seroprevalence of and risk factors for feline coronavirus infection in cats from Greece.

Feline coronavirus (FCoV) is a highly contagious and ubiquitous virus of domestic cats and wild felids. Feline infectious peritonitis (FIP) is a fatal, systemic disease caused by FCoV infection when spontaneous mutations of the viral genome take place. The aims of this study were primarily to determine the prevalence of seropositivity for FCoV in different populations of cats in Greece and assess risk factors for seropositivity. A total of 453 cats were prospectively enrolled in the study. A commercially available IFAT kit was used for the detection of FCoV IgG antibodies in serum. Overall, 55 (12.1 %) of the 453 cats were seropositive for FCoV. Based on multivariable analysis, factors associated with FCoV-seropositivity included cats adopted as strays and contact with other cats. This is the first extensive study on the epidemiology of FCoV in cats from Greece and one of the largest worldwide. Feline coronavirus infection is relatively common in Greece. Therefore, it is necessary to establish optimal strategies for the prevention of FCoV infection, considering the high-risk groups of cats identified in this study.

Investigation of the diagnostic performance of prick test with two environmental allergens in dogs with atopic dermatitis: A study using intradermal test as the gold standard.

BACKGROUND: There are no studies investigating the correlation between prick test (PT) and intradermal test (IDT) with environmental allergens in dogs with atopic dermatitis (AD). OBJECTIVES: To investigate the correlation between PT and IDT for two environmental allergens, and to calculate the sensitivity, specificity and Youden index of PT, using IDT as the gold standard. MATERIALS AND METHODS: Twenty-two dogs with AD were selected. PT was performed with glycerinated allergen extracts, along with negative and positive controls, using the Greer Prick System. Reactions were interpreted (positive/negative) subjectively and by using seven objective criteria, by an examiner blinded to the IDT results. IDT reactions to the same allergens were interpreted, subjectively and objectively, by another masked investigator. The agreement between PT and IDT, the sensitivity, specificity and Youden index of PT, using IDT as gold standard, were calculated. RESULTS: On subjective evaluation, the correlation between PT and IDT was poor and sensitivity of PT was 0%. Of the seven criteria for the objective evaluation of PT, the best diagnostic performance was attained when allergen-induced wheals were considered positive if their longest diameter was ≥8.5 mm. However, even then, the correlation with IDT was moderate, and the sensitivity of PT, albeit based on few positive IDT reactions, was low. CONCLUSION AND CLINICAL RELEVANCE: At least as performed herein, PT has a poor-to-moderate correlation with IDT, mainly as a consequence of the lack of positive PT reactions. Further studies are needed to improve PT technique, yet, meanwhile, it cannot be recommended as a substitute for IDT.

Repeatability and reproducibility of the interpretation of intradermal test results in dogs with atopic dermatitis.

Background - There is lack of studies evaluating the repeatability and reproducibility of the interpretation of intradermal testing in dogs with atopic dermatitis (AD). Objectives - To evaluate the repeatability and reproducibility of the interpretation of intradermal test results in dogs with AD. For comparison, the repeatability of allergen-specific immunoglobulin (Ig)E serology also was examined. Materials and methods - Twenty dogs with AD were used. Intradermal test included injections of known negative and positive controls, and of 25 masked injections of 10 allergens/controls, that were selected randomly and injected at random positions. Reactions to the 25 masked allergens/controls were scored (positive/negative) subjectively by three independent examiners followed by an objective assessment. Allergen-specific IgE serology was performed in blinded duplicate samples collected from all dogs for nine of 10 of the same allergens. Results - Kappa values of intraobserver repeatability (≥2 injections of the same allergen to different positions of the same dog) varied between -0.53 and 0.8 (subjective evaluations), and between 0.03 and 1 (objective evaluation). When the repeatability of the serological test was examined k = 0.91. Kappa values for the interobserver reproducibility (objective and three subjective evaluations of the same allergen injected at the same position of the same dog) varied between 0.6 and 0.74 (overall 0.67). Conclusions and clinical relevance - Intraobserver repeatability of the subjective and objective evaluation of IDT results varied from good to poor and depended on the number of times the same allergen was injected, whereas interobserver reproducibility varied from substantial to moderate. Further studies are needed to optimise the repeatability and reproducibility of IDT in dogs.

Contexte - Il existe un manque d'études évaluant la répétabilité et la reproductibilité de l'interprétation des tests intradermiques chez les chiens atteints de dermatite atopique (DA). Objectifs - Évaluer la répétabilité et la reproductibilité de l'interprétation des résultats des tests intradermiques chez les chiens atteints de MA. À des fins de comparaison, la répétabilité de la sérologie de l'immunoglobuline (Ig)E spécifique de l'allergène a également été examinée. Matériels et méthodes - Vingt chiens atteints de MA ont été utilisés. Le test intradermique comprenait des injections de contrôles négatifs et positifs connus, et de 25 injections masquées de 10 allergènes/contrôles, qui ont été sélectionnés au hasard et injectés à des positions aléatoires. Les réactions aux 25 allergènes/contrôles masqués ont été notées (positives/négatives) subjectivement par trois examinateurs indépendants, suivies d'une évaluation objective. La sérologie IgE spécifique de l'allergène a été réalisée dans des échantillons en double en aveugle prélevés sur tous les chiens pour neuf des 10 allergènes identiques. Résultats - Les valeurs kappa de répétabilité intra-observateur (≥2 injections du même allergène à différentes positions du même chien) variaient entre -0,53 et 0,8 (évaluations subjectives), et entre 0,03 et 1 (évaluation objective). Lorsque la répétabilité du test sérologique a été examinée, k = 0,91. Les valeurs de Kappa pour la reproductibilité interobservateur (évaluations objective et trois évaluations subjectives du même allergène injecté au même endroit du même chien) variaient entre 0,6 et 0,74 (globalement 0,67). Conclusions et pertinence clinique - La répétabilité intra-observateur de l'évaluation subjective et objective des résultats IDT variait de bonne à mauvaise et dépendait du nombre d'injections d'un même allergène, alors que la reproductibilité inter-observateur variait de substantielle à modérée. D'autres études sont nécessaires pour optimiser la répétabilité et la reproductibilité de l'IDT chez le chien.

Introducción- faltan estudios que evalúen la repetibilidad y la reproducibilidad de la interpretación de las pruebas intradérmicas en perros con dermatitis atópica (AD). Objetivos- evaluar la repetibilidad y reproducibilidad de la interpretación de los resultados de las pruebas intradérmicas en perros con AD. A modo de comparación, también se examinó la repetibilidad de la serología de inmunoglobulina (Ig)E específica de alérgeno. Materiales y métodos - Se utilizaron 20 perros con AD. La prueba intradérmica incluyó inyecciones de controles positivos y negativos conocidos, y de 25 inyecciones enmascaradas de 10 alérgenos/controles, que se seleccionaron al azar y se inyectaron en posiciones aleatorias. Las reacciones a los 25 alérgenos/controles enmascarados fueron calificadas (positivas/negativas) subjetivamente por tres examinadores independientes seguido de una evaluación objetiva. La serología de IgE específica para alérgenos se realizó en muestras duplicadas ciegas recolectadas de todos los perros para nueve de 10 de los mismos alérgenos. Resultados - Los valores Kappa de repetibilidad intraobservador (≥2 inyecciones del mismo alérgeno en diferentes posiciones del mismo perro) variaron entre -0,53 y 0,8 (evaluaciones subjetivas) y entre 0,03 y 1 (evaluación objetiva). Cuando se examinó la repetibilidad de la prueba serológica k = 0,91. Los valores de Kappa para la reproducibilidad interobservador (objetivo y tres evaluaciones subjetivas del mismo alérgeno inyectado en la misma posición del mismo perro) variaron entre 0,6 y 0,74 (en general, 0,67). Conclusiones y relevancia clínica- la repetibilidad intraobservador de la evaluación subjetiva y objetiva de los resultados de la IDT varió de buena a mala y dependió del número de veces que se inyectó el mismo alérgeno, mientras que la reproducibilidad interobservador varió de sustancial a moderada. Se necesitan más estudios para optimizar la repetibilidad y reproducibilidad de IDT en perros.

Contexto - Há poucos estudos avaliando a repetibilidade e reprodutibilidade da interpretação do teste intradérmico em cães com dermatite atópica (DA). Objetivos - Avaliar a repetibilidade e reprodutibilidade da interpretação dos resultados de testes intradérmicos em cães com DA. Para comparação, a repetibilidade da sorologia com imunoglobulinas (Ig)E alérgeno-específicas foi também avaliada. Materiais e métodos - Foram utilizados 20 cães com DA. O teste intradérmico incluiu injeções de controles negativos e positivos conhecidos e de 25 injeções mascaradas de 10 alérgenos/controles, que foram selecionados aleatoriamente e injetadas em posições aleatórias. As reações aos 25 alérgenos/controles mascarados foram pontuadas (positiva/negativa) subjetivamente por três examinadores independentes, seguidas de uma avaliação objetiva. A sorologia de IgE específica para alérgenos foi realizada em amostras duplicadas cegas coletadas de todos os cães para nove de 10 dos mesmos alérgenos. Resultados - Os valores Kappa de repetibilidade intraobservador (≥2 injeções do mesmo alérgeno em diferentes posições do mesmo cão) variaram entre -0,53 e 0,8 (avaliação subjetiva) e entre 0,03 e 1 (avaliação objetiva). Quando examinada a repetibilidade do teste sorológico k=0,91. Os valores de Kappa para a reprodutibilidade interobservador (objetiva e três avaliações subjetivas do mesmo alérgeno injetado na mesma posição do mesmo cão) variaram entre 0,6 e 0,74 (total 0,67). Conclusões e relevância clínica - A repetibilidade intraobservador da avaliação subjetiva e objetiva dos resultados do IDT variou de boa a ruim e dependeu do número de vezes que o mesmo alérgeno foi injetado, enquanto a reprodutibilidade interobservador variou de substancial a moderada. Mais estudos são necessários para otimizar a repetibilidade e reprodutibilidade do IDT em cães.

Serial Measurement of Serum Pancreatic Lipase Immunoreactivity, Feline Trypsin-like Immunoreactivity, and Cobalamin Concentrations in Kittens.

Serum concentrations of feline pancreatic lipase immunoreactivity (fPLI), feline trypsin-like immunoreactivity (fTLI), and cobalamin are commonly used for the diagnostic investigation of cats with gastrointestinal signs. No information on these parameters in healthy cats less than 1 year of age exists. We aimed to evaluate serum concentrations of fPLI, fTLI, and cobalamin in healthy cats at different time-points during their first 12 months of life. Fourteen healthy 2-month-old kittens were included. Blood was collected at 2, 3, 4, 6, and 12 months of age, and serum concentrations of fPLI, fTLI, and cobalamin were measured. While there was a statistically significant difference in serum fPLI concentrations over time, there was no statistically significant difference between individual time-points. There was no significant difference in serum fTLI concentrations over time. Serum cobalamin concentrations were below the reference interval in 3/13 cats at 2 months of age and were significantly lower by 3 months, when 13/14 had hypocobalaminemia. By 12 months, serum cobalamin had significantly increased, yet 4/12 cats still had hypocobalaminemia. Serum fPLI and fTLI concentrations did not show any statistically or clinically significant differences in young kittens. In contrast, serum cobalamin concentrations were commonly below the reference interval in kittens. Serum fPLI and fTLI concentrations are not practically affected by age in kittens as young as 2 months of age and could be used for the investigation of pancreatic diseases.

Prevalence and Risk Factors for Bartonella spp. and Haemoplasma Infections in Cats from Greece.

Bartonellosis and haemoplasmosis are vector-borne diseases with global impact on the health of domestic cats and of zoonotic importance. The aim of this study was to describe the epidemiological aspects of various populations of cats infected with Bartonella spp. or haemoplasma species. The populations evaluated included client-owned cats, stray cats and cats that live in breeding catteries in Greece. A total of 452 cats were prospectively enrolled into the study. A commercially available indirect immunofluorescence antibody testkit was used for the detection of Bartonella henselae IgG antibodies in serum. PCRs for the detection of Bartonella spp. and haemoplasma species DNA in the blood were also performed in a subgroup of 242 of the 452 cats. Risk factors for B. henselae seropositivity and infection with the haemoplasma species were determined using multivariable analysis. Overall, 160 (35.4%) of the 452 cats were seropositive for B. henselae. Seven (2.9%) and 46 (19%) of the 242 cats were PCR-positive for Bartonella spp. and haemoplasma species, respectively. The factors associated with B. henselae seropositivity, based on multivariate analysis, included older age, outdoor access, living region and flea infestation. Non-administration of ectoparasiticides was associated with haemoplasma species infection. This study shows a high prevalence of seropositivity for B. henselae and a relatively high prevalence of infection with haemoplasma species. Therefore, it is necessary to establish optimal strategies for the prevention of Bartonella spp. and haemoplasma species infections, considering the high-risk groups of cats identified in this study.

Prevalence of antinuclear antibodies and rheumatoid factor titers in dogs with arthritis secondary to leishmaniosis (Leishmania infantum).

Dogs with infectious arthritis may occasionally exhibit positive serum antinuclear antibody (ANA) and rheumatoid factor (RF) titers; however, relevant data are sparse for arthritis secondary to canine leishmaniosis (CanL) caused by Leishmania infantum. We determined the prevalence of positive serum ANA and RF titers in dogs with arthritis secondary to CanL. Blood samples from adult, client-owned dogs with purulent arthritis secondary to CanL, without any comorbidities, were collected for diagnostic purposes. Serum ANA titers were measured by immunoperoxidase test and RF titers by the Rose-Waaler latex test. Twelve of 23 dogs enrolled prospectively in our study had clinical arthritis, and 11 of 23 had subclinical arthritis. Based on LeishVet clinical staging, 7 dogs had clinical stage II disease, 11 had clinical stage III disease, and 5 had stage IV. None of the 23 dogs was seropositive for ANA; 3 of 23 were positive for RF. ANA and/or RF seropositivity, in dogs with CanL-associated arthritis, appears to be weak, if present at all. Based on our results, positive serum ANA and RF titers should not be expected in dogs with arthritis secondary to CanL.

Repeatability and reproducibility of microscopic examination of adhesive tape strip cytology slides for the quantification of Malassezia spp. in canine skin.

BACKGROUND: The optimal microscopic magnification and number of optical fields of adhesive tape strip cytological slides that should be examined when searching for Malassezia yeasts on canine skin are unknown. OBJECTIVES: To determine the optimal magnification and the minimum number of optical fields that should be examined to maximise intraobserver repeatability and interobserver reproducibility. MATERIALS AND METHODS: Seven experienced examiners counted, twice, the number of yeasts in 10, 20, 30, 40 and 50 optical fields of 40 slides at ×400 and ×1000 magnification. RESULTS: The number of yeasts per unit surface area was significantly higher at ×1000 compared to ×400 magnification. Repeatability and reproducibility for counting the yeasts was very poor. CONCLUSIONS AND CLINICAL RELEVANCE: Adhesive tape strip cytological slides should be examined microscopically for Malassezia spp. at ×1000 magnification. The repeatability of this examination for counting the yeasts is poor.

Contexte - Le grossissement microscopique optimal et le nombre de champs optiques des lames cytologiques de bandes adhésives à examiner lors de la recherche de levures Malassezia sur la peau de chien sont inconnus. Objectifs - Déterminer le grossissement optimal et le nombre minimal de champs à examiner pour maximiser la répétabilité intra-observateur et la reproductibilité inter-observateur. Matériels et méthodes - Sept examinateurs expérimentés ont compté, deux fois, le nombre de levures dans 10, 20, 30, 40 et 50 champs de 40 lames aux grossissements ×400 et ×1 000. Résultats - Le nombre de levures par unité de surface était significativement plus élevé au grossissement ×1 000 par rapport au grossissement ×400. La répétabilité et la reproductibilité du comptage des levures étaient très médiocres. Conclusions et pertinence clinique - Les lames cytologiques de bandes adhésives doivent être examinées au microscope pour Malassezia spp. à un grossissement ×1 000. La répétabilité de cet examen de comptage des levures est faible.

Introducción- se desconoce el aumento microscópico óptimo y el número de campos ópticos de los portaobjetos citológicos en tiras de cinta adhesiva que deben examinarse al buscar levaduras Malassezia en la piel canina. Objetivos- determinar el aumento óptimo y el número mínimo de campos ópticos que deben examinarse para maximizar la repetibilidad intraobservador y la reproducibilidad interobservador. Materiales y métodos- siete examinadores experimentados contaron dos veces el número de levaduras en campos ópticos de 10, 20, 30, 40 y 50 de 40 portaobjetos con aumentos de x ×400 y ×1000. Resultados- el número de levaduras por unidad de superficie fue significativamente mayor con un aumento de ×1000 en comparación con un aumento de ×400. La repetibilidad y reproducibilidad para contar las levaduras fue muy pobre. Conclusiones y relevancia clínica - Los portaobjetos citológicos en tiras de cinta adhesiva deben examinarse microscópicamente para detectar Malassezia spp. con un aumento de ×1.000. La repetibilidad de este examen para contar las levaduras es pobre.

Contexto - A ampliação microscópica ideal e o número de campos ópticos das lâminas citológicas de fita adesiva que devem ser examinados nas pesquisas de leveduras do gênero Malassezia em cães são desconhecidos. Objetivos - Determinar a magnificação ideal e o número mínimo de campos ópticos que devem ser examinados para maximizar a repetibilidade intraobservador e a reproducibilidade interobservador. Materiais e métodos - Sete examinadores experientes contaram duas vezes o número de leveduras em 10, 20, 30, 40 e 50 campos ópticos de 40 lâminas nas magnificações de x400 e x1000. Resultados - O número de leveduras por unidade de área de superfície foi significativamente maior em x1000 em comparação com a ampliação de x400. A repetibilidade e a reprodutibilidade para a contagem de leveduras foi muito pobre. Conclusões e relevância clínica - Lâminas de citologia por fica adesiva devem ser examinadas microscopicamente para Malassezia spp a uma magnificação de x1.000. A repetibilidade deste exame para contagem de leveduras foi pobre.

Excitement-Induced Cutaneous Bleeding (Haematidrosis-like) in a Dog.

A 15-month-old intact female Pitbull was referred because of recurrent, episodic, self-limiting, excitement-induced bleeding from nontraumatised skin. No abnormalities were detected upon physical examination. Subsequently, the dog went for a walk under the direct supervision of one of the authors, became overexcited and, after approximately five minutes, bloody liquid, with a patchy distribution, appeared along the hair shafts of the face and neck. The affected skin was congested, partially blanching on diascopy and bloody liquid was oozing from the follicular openings. Urticaria, dermographism and hypertension were excluded, the complete blood count and coagulation profile were within the reference ranges and an analysis of the bloody exudate confirmed its blood components. The cutaneous bleeding of the dog followed a self-limited course, with no episodes during the last two years. Clinical and laboratory findings and the long-term evolution of this dog bear striking similarities to haematidrosis, a rare human disease of multifactorial aetiology and equivocal pathogenesis.

A proposed medication score for long-term trials of treatment of canine atopic dermatitis sensu lato.

BACKGROUND: The use of concurrent medications is necessary in trials of treatment of canine atopic dermatitis. Our aim was to use the best available evidence to construct and then to validate a medication score (MS) formula that will estimate the impact of concurrent medications on trial outcomes. METHODS: Trials of 15 interventions were scrutinized to find those that were consistent in terms of specific medication, administration route and dosage regimen. A MS was constructed in five steps, starting from assigning a score of 1 for each day on oral prednisone, prednisolone or methylprednisolone at 0.5-1.0 mg/kg. The MS score was validated using the clinical records of 35 dogs with atopic dermatitis that had been treated for a period of 12 ± 2 weeks with six of these medications and compared with a previously published non-validated MS. RESULTS: A MS could be assigned to eight treatments, six of which had been administered to the 35 dogs. A positive correlation was seen with the previously published MS and a negative correlation with changes in lesional and pruritus scores. CONCLUSION: This MS may be a useful tool in new studies evaluating the efficacy of treatments in canine atopic dermatitis.

Evaluation of clinicopathological abnormalities in sick cats naturally infected by Leishmania infantum.

Feline infection by Leishmania infantum (syn. L. chagasi) has been described in areas where canine leishmaniosis is endemic. A wide variety of clinicopathological abnormalities have been reported in cats presenting clinical signs of leishmaniosis but there is a paucity of information regarding cats infected by L. infantum that do not suffer from leishmaniosis but from other diseases. The aim of this study was to compare: a) the frequency of clinicopathological abnormalities and b) the values of hematology, serum biochemistry and urinalysis parameters, between non-infected sick cats and sick cats that were infected by L. infantum. A total of 50 cats with cutaneous, ocular and/or systemic clinical signs that lived in an endemic area and had been tested for infection by L. infantum using PCR from four different tissues, were included. Based on the results of PCR, 20/50 cats were found to be infected and 30/50 non-infected. The only difference between the two groups of cats was that the concentration of inorganic phosphorus (P = 0.043) was higher in infected cats. This finding may suggest an association between infection by L. infantum and feline kidney disease.

Evaluation of serum symmetric dimethylarginine as a biomarker of kidney disease in canine leishmaniosis due to Leishmania infantum.

Canine leishmaniosis (CanL)-associated chronic kidney disease is a leading cause of morbidity and mortality in Mediterranean countries. Novel renal biomarkers, such as serum symmetric dimethylarginine (sSDMA), may be useful surrogates for the detection of renal functional impairment. The objectives of this study were to investigate sSDMA concentrations in dogs with CanL, with and without azotemia, and to establish any potential association with the prevalence and severity of proteinuria, with the prevalence of decreased urine specific gravity and with the LeishVet clinical stages of CanL. Serum samples from 68 dogs with CanL (50 nonazotemic and 18 azotemic) and 17 healthy dogs were retrospectively examined. Increased sSDMA was documented in 26 % of dogs with CanL without azotemia and in 83.3 % of dogs with azotemia. Serum SDMA was significantly higher in azotemic compared to nonazotemic dogs and was associated with the presence and severity of proteinuria, the decreased urine specific gravity and the advanced clinical stages of CanL. The results of the present study indicate that sSDMA may be a useful adjunct to serum creatinine and urine protein/creatinine ratio for the detection of CanL-associated nephropathy, but it is of limited value for distinguishing among the LeishVet clinical stages of CanL.

The lipid profile of three Malassezia species assessed by Raman spectroscopy and discriminant analysis.

Malassezia yeasts constitute the major eukaryotic cutaneous flora of homoeothermic vertebrates. These lipophilic yeasts are able to cause, trigger, or aggravate common skin diseases under favorable conditions. Species identification and subspecies differentiation is currently based on morphological characteristics, lipid assimilation profile, and molecular tests. Mass spectrometry has been also reported as a reliable, yet costly and labor-intensive, method to classify Malassezia yeasts. Here, we introduce Raman spectroscopy as a new molecular technique able to differentiate three phylogenetically close Malassezia species (M.globosa, M.pachydermatis, and M.sympodialis) by examining their lipid metabolic profile. Using Raman spectroscopy, lipid fingerprints of Malassezia cultures on Leeming-Notman agar, were analyzed by spectral bands assignment and partial least squares discriminant analysis. Our results demonstrate differential utilization of lipid supplements among these three species and the ability of Raman spectroscopy to rapidly and accurately discriminate them by predictive modelling.

Prevalence and mechanisms of resistance to fluoroquinolones in Pseudomonas aeruginosa and Escherichia coli isolates recovered from dogs suffering from otitis in Greece.

The aim of this study was to investigate the prevalence and the implicated mechanisms of resistance against selected veterinary fluoroquinolones (enrofloxacin, marbofloxacin and pradofloxacin) among 101 Pseudomonas aeruginosa (n=75) and Escherichia coli (n=26) isolates collected from dogs suffering from otitis. Resistance ranged from 32.0% to 48.0% with differences not being considered statistically significant among the three agents or between the two bacterial species. However, individual MICs of pradofloxacin, the latest veterinary fluoroquinolone, were significantly lower than those of enrofloxacin, the oldest one, indicating an increased in vitro potency of the former antimicrobial. Pradofloxacin MIC90 was, additionally, the lowest (8µg/ml), in E. coli, or among the lowest (8µg/ml), in P. aeruginosa isolates. Resistance was in most cases associated with topoisomerase substitutions, with patterns GyrA:V73G in P. aeruginosa and GyrA:S83L+D87N/ParC:S58I+A86V in E. coli being reported for the first time in small animal isolates. Only 6.7% and 15.4% of P. aeruginosa and E. coli otitis isolates, respectively, carried plasmid-mediated quinolone resistance (PMQR) genes, which, moreover, contributed minimally to resistance. Efflux pump activity was additionally detected in resistant E. coli isolates, even those lacking topoisomerase substitutions or PMQR genes. The emergence of resistance in the canine otitis isolates seemed to be associated with previous, prolonged systemic fluoroquinolone administration. In any case, antimicrobial susceptibility testing should guide the selection of systemic FQs for the treatment of canine otitis.

Molecular detection of vector-borne pathogens in Greek cats.

Infectious diseases have been increasingly recognized in cats worldwide. The objective of this study was the molecular investigation of the prevalence of selected pathogens in healthy and sick cats from Greece, a country highly endemic for several canine vector-borne pathogens. Blood and/or bone marrow samples from 50 clinically healthy and 50 sick adult (>1 year-old) cats were retrospectively examined for the amplification of Bartonella spp., haemoplasmas, Ehrlichia spp., Anaplasma spp., Babesia spp., and Cytauxzoon spp. DNA. Overall, 14.9% of the cats were found to be infected or co-infected by haemoplasmas, including Candidatus Mycoplasma haemominutum and M. haemofelis. In addition, 8.5% of the cats were infected by Bartonella henselae, Bartonella clarridgeiae or Bartonella koehlerae. In contrast, DNA of Ehrlichia spp., Anaplasma spp., Babesia spp. and Cytauxzoon spp. was not amplified from the blood or bone marrow of any cat. There was no significant difference in either haemoplasma or Bartonella infection rates when comparing healthy and sick cats. This study represents the first description of Bartonella koehlerae in Greek cats.

A field trial of a fixed combination of permethrin and fipronil (Effitix®) for the treatment and prevention of flea infestation in dogs living with sheep.

BACKGROUND: A large number of fleas parasitize dogs living with sheep in Greece. The primary aim of this randomized, blinded, placebo-controlled trial was to examine the efficacy of a permethrin-fipronil combination (Effitix®) for the treatment and prevention of flea infestation in dogs living with sheep and the secondary aim was to examine the efficacy of this intervention on flea infestation, pruritus and skin lesions of the people in contact with these dogs. METHODS: Thirty dogs living with sheep and infested by at least 10 fleas and all 80 sheep living on the same premises were randomly allocated into equal groups. Group A dogs were treated three times, every 4 weeks, with a spot-on containing 54.5% permethrin and 6.1% fipronil, group A sheep were treated, on the same days, with a pour-on containing 1% deltamethrin, whereas group B dogs were sham-treated and group B sheep were placebo-treated. Flea counting was performed at the beginning of the trial (day 0) and after 14, 28, 56 and 84 days and the first five fleas from each animal were used for species identification. At the same time points, flea infestation, pruritus and skin lesions of the people in contact with the dogs were assessed. RESULTS: The percentage of dogs with zero flea counts was significantly higher in group A than in group B on days 14, 28, 56 and 84 and flea counts were significantly lower in group A dogs than in group B dogs at the same time points. The percent efficacy of the permethrin-fipronil combination was higher than 78% (arithmetic means) or than 96% (geometric means) throughout the study. No adverse reactions were recorded. Between the two flea species found on dogs, Ctenocephalides canis was predominant over C. felis. Flea-infected sheep were not found at the beginning or during the study and no significant changes in flea infestation, pruritus and skin lesions of the people in contact with the dogs were witnessed throughout the study. CONCLUSIONS: A spot-on solution containing 54.5% permethrin and 6.1% fipronil is safe and effective for the treatment and prevention of C. canis and C. felis infestations in dogs living with sheep.