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.

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.

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.

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.

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.

A field trial of spinosad for the treatment and prevention of flea infestation in shepherd dogs living in close proximity to flea-infested sheep.

BACKGROUND: Three flea species, Pulex irritans, Ctenocephalides canis and C. felis parasitize shepherd dogs living on sheep farms in Greece. The aim of this randomized, blinded, placebo-controlled trial was to examine the efficacy of spinosad, when administered three times every 4 weeks, as the only intervention to treat and prevent flea infestations in shepherd dogs living on sheep farms. METHODS: Thirty dogs living on sheep farms and infested by at least 24 fleas were randomly allocated into equal groups. Group A dogs received spinosad (45-70 mg/kg body weight), every 4 weeks for three administrations, whereas Group B dogs were placebo-treated. Flea counting was performed at the beginning of the trial (day 0) and after 14, 28, 56 and 84 days. The first five fleas from each dog and 2-6 fleas collected from 5-11 sheep were used for species identification. RESULTS: The percentage of dogs with zero flea counts was significantly higher in group A than in group B at days 14, 28, 56 and 84 and flea counts were significantly lower in group A than in group B at days 14, 28, 56 and 84. In group A, flea counts were significantly lower at days 14, 28, 56 and 84 compared to day 0 whereas there were no changes in flea counts of group B dogs. The percent efficacy of spinosad for the treatment and prevention of flea infestation was higher than 98% (arithmetic means) or higher than 99% (geometric means) throughout the study. No adverse reactions were recorded. C. canis was the predominant flea species of dogs at day 0. In group A the relative abundance of C. felis increased at day14 whereas in group B the relative abundance of P. irritans increased at days 14, 28, 56 and 84. CONCLUSIONS: Spinosad is safe and effective for the treatment of C. canis and C. felis infestations and for the prevention of P. irritans, C. canis and C. felis infestations in shepherd dogs living in close proximity to sheep.

Evaluation of indirect immunofluorescence antibody test and enzyme-linked immunosorbent assay for the diagnosis of infection by Leishmania infantum in clinically normal and sick cats.

Cats that live in areas where canine and human leishmaniosis due to Leishmania infantum is endemic may become infected and may develop anti-Leishmania antibodies. In this study 50 clinically normal and 50 cats with cutaneous and/or systemic signs that lived in an endemic area and had been previously examined for infection by L. infantum using PCR in four different tissues were serologically tested for the presence of anti-Leishmania IgG (IFAT and ELISA) and IgM (IFAT). The aim was to compare the results of IFAT, ELISA and PCR and to investigate the possible associations between seropositivity to Leishmania spp and signalment, living conditions, season of sampling, health status of the cats, and seropositivity to other infectious agents. Low concentrations of anti-Leishmania IgG were detected by IFAT in 10% of the cats and by ELISA in 1%, whereas anti-Leishmania IgM were detected by IFAT in 1%. There was disagreement between the results of IFAT and ELISA for anti-Leishmania IgG (P = 0.039) and between all serological tests and PCR (P < 0.001). The diagnostic sensitivity of all serological tests, using PCR as the gold standard, was very low, but ELISA and IFAT for anti-Leishmania IgM had 100% specificity. The diagnostic sensitivity of all serological tests could not be improved by changing the cut-off values. Seropositivity for Leishmania spp was not associated with signalment, living conditions, season of sampling and health status of the cats or with seropositivity to feline leukemia virus, feline immunodeficiency virus, feline coronavirus, Toxoplasma gondii and Bartonella henselae. In conclusion, because of their low sensitivity and very high specificity two of the evaluated serological tests (ELISA for anti-Leishmania IgG and IFAT for anti-Leishmania IgM) may be useless as population screening tests but valuable for diagnosing feline infection by L. infantum.

Prospective evaluation of serum pancreatic lipase immunoreactivity and troponin I concentrations in Leishmania infantum-infected dogs treated with meglumine antimonate.

Canine leishmaniosis (CanL) caused by Leishmania infantum is an important zoonotic disease. One of the most commonly used drugs for the treatment of CanL is meglumine antimonate. Drugs of this class have been associated with pancreatitis and cardiotoxicity in humans infected with Leishmania spp. The aim of this study was to measure serum canine pancreatic lipase immunoreactivity (Spec cPL) and cardiac troponin I (cTnI) concentrations in dogs with leishmaniosis during treatment with meglumine antimonate, and to compare them with those of dogs with leishmaniosis not treated with this drug. A total of 30 non-uremic dogs with leishmaniosis, living in Greece, were prospectively enrolled into the study. Of the 30 dogs, 20 (Group A) were treated with a combination of meglumine antimonate (100mg/kg, SC, q24 h) and allopurinol (10mg/kg, PO, q12h) for 28 days, while 10 dogs (Group B) were treated with allopurinol alone (10mg/kg, PO, q12h) for 28 days. Blood samples were collected at timepoint 0 (before treatment) and at 14 and 28 days after the initiation of treatment. None of the dogs treated with meglumine antiomonate had a Spec cPL concentration suggestive of pancreatitis (≥ 400 µg/L) or clinical signs suggestive of pancreatitis at any of the timepoints. Similarly, none of the dogs treated with meglumine antiomonate had a serum cTnI concentration above the upper limit of the reference range (>0.5 ng/mL) or clinical evidence of cardiotoxicity at any of the 3 timepoints. In the present study, meglumine antimonate treatment in dogs with leishmaniosis did not result in clinical or laboratory evidence of either pancreatitis or cardiotoxicity.

Cytological and molecular detection of Leishmania infantum in different tissues of clinically normal and sick cats.

Natural infection of domestic cats by Leishmania infantum (synonym: L. chagasi) has been demonstrated in several European, Latin American, and Asian countries, and the estimated prevalence of infection, based mainly on blood PCR, ranges from 0.3% up to 60.6%. In this study we aimed to: (a) estimate the prevalence of the infection by L. infantum in clinically normal cats (group A; n=50) and in cats with various clinical signs (group B; n=50), living in an endemic region, by both cytological examination of four different tissues (lymph node, skin, bone marrow, and conjunctiva) and by PCR in four different tissues (blood, skin biopsies, bone marrow, and conjunctiva); (b) compare the diagnostic sensitivity of the above methods and evaluate for possible associations between their results; and (c) investigate the possible associations between infection by L. infantum and signalment, living conditions, season of sampling, and health status of the cats. The prevalence of the infection in the study population was 41% and did not differ (P=0.839) between group A (42%) and B (40%) cats. Lymph node, skin, bone marrow and conjunctiva cytology was always negative. Therefore, the diagnosis of the infection was based only on PCR in blood, skin biopsy, bone marrow and conjunctiva, which was positive in 13%, 18.2%, 16% and 3.1% of the cats, respectively. PCR was positive in only one of the four tissues in 80.5% of the infected cats. The results differed (P=0.014) among the four tissues and were less frequently positive in conjunctiva compared to skin biopsies and bone marrow (P=0.007 for both comparisons), thus highlighting the need for multiple tissue PCR testing in order to minimize false-negative results. More PCR-positive cats were found when sampling was performed during the period of sandfly activity (odds ratio: 2.44; P=0.022). Also, in group B cats, the likelihood of PCR-positivity was higher (odds ratio: 3.93; P=0.042) among those presenting at least one systemic clinical sign that had been previously reported in cats with leishmaniosis.

Comparison of two commercial rapid in-clinic serological tests for detection of antibodies against Leishmania spp. in dogs.

Antibodies against Leishmania spp. are detected in most dogs with clinical signs of leishmaniasis due to Leishmania infantum. Accurate, rapid in-clinic serological tests may permit immediate confirmation of the diagnosis and implementation of therapeutic measures. The aim of the current study was to evaluate the diagnostic accuracy of 2 commercial, rapid in-clinic serological tests for the detection of anti-Leishmania antibodies in sera of dogs, the Snap Canine Leishmania Antibody Test kit (IDEXX Laboratories Inc., Westbrook, Maine) and the ImmunoRun Antibody Detection kit (Biogal Galed Labs, Kibbutz Galed, Israel), using indirect fluorescent antibody test (IFAT) as the reference method. A total of 109 sera collected from 65 seropositive and 44 seronegative dogs were used. The sensitivities of the Snap and ImmunoRun kits were 89.23% (95% confidence interval: 79.05-95.54%) and 86.15% (95% confidence interval: 75.33-93.45%), respectively, and the specificity of both tests was 100%. A good agreement between each of the rapid in-clinic serological tests and IFAT and between the 2 rapid in-clinic serological tests was witnessed. Both rapid in-clinic serological tests showed an adequate diagnostic accuracy and can be used for the fast detection of antibodies against L. infantum in dogs.

Thyroid function in 36 dogs with leishmaniosis due to Leishmania infantum before and during treatment with allopurinol with or without meglumine antimonate.

Hypothyroidism may predispose to the development of canine leishmaniosis or it may appear during the course of the latter due to infiltration and destruction of the thyroid gland by infected macrophages. The main purpose of this study was to evaluate thyroid function through measurement of serum total thyroxin (tT4), free thyroxin (fT4), and canine thyroid stimulating hormone (cTSH) concentrations in 36 dogs with leishmaniosis, before and after 2 and 4 weeks of treatment with allopurinol with or without meglumine antimonate. Before treatment 27/36 (75%) dogs had serum tT4 concentrations below the lower limit of the reference interval but only 2 of them had concurrently serum fT4 concentrations below the lower limit of the reference interval and none had increased serum cTSH concentrations. During treatment there were no significant changes in serum tT4 or fT4 concentrations, whereas a significant increase in serum cTSH was observed. Two dogs had decreased serum tT4 and fT4 but normal cTSH concentrations before treatment and two other dogs had decreased serum tT4 and increased cTSH, but normal fT4 concentrations during the treatment period. Although hypothyroidism could not be definitively excluded in these dogs it is considered unlikely based on their overall hormonal profile, clinical presentation, and response to treatment. Therefore, hypothyroidism does not appear to be an important predisposing disease or a frequent complication of canine leishmaniosis.

Concentrations of clindamycin hydrochloride in homogenates of normal dog skin when administered at two oral dosage regimens.

BACKGROUND: Clindamycin is frequently used for the treatment of bacterial pyoderma. OBJECTIVE: To compare the pharmacokinetics of clindamycin in whole skin homogenates of normal dogs when administered orally at two dosage regimens (5.5 mg/kg BW, twice daily and 11 mg/kg BW, once daily). ANIMALS AND METHODS: Skin biopsies were obtained from six laboratory beagles before, 3, 6 and 12 h after the first and the fifth dose of clindamycin at the former regimen, as well as before, 3, 6, 12 and 24 h after the first and third dose at the latter regimen. Tissue was homogenized and clindamycin concentrations were measured by reverse-phase high-performance liquid chromatography coupled with mass spectrometry. Results were analyzed using Student's t-test at a level of significance of 0.05. RESULTS: Maximal concentration and area under the concentration-time curve, but not their relevant dose-normalized values, were higher at the dosage regimen of 11 mg/kg BW, once daily than at 5.5 mg/kg BW twice daily. CONCLUSIONS: The pharmacokinetic profile of clindamycin is at least equal, if not better, when this antimicrobial is administered at 11 mg/kg BW, once daily.

Serum pharmacokinetics of clindamycin hydrochloride in normal dogs when administered at two dosage regimens.

The aim of this cross-over study was to compare clindamycin pharmacokinetics in the serum of clinically normal dogs when administered orally at two dosage regimens (5.5 mg/kg, twice daily, and 11 mg/kg, once daily), separated by a 1 week wash-out period. Serum samples were obtained from six clinically normal laboratory beagles before, 3, 6, 9 and 12 h after the first and fifth dose of clindamycin at 5.5 mg/kg, twice daily, and before, 3, 6, 9, 12, 18 and 24 h after the first and third dose at 11 mg/kg, once daily. Serum clindamycin concentrations were determined by reverse-phase liquid chromatography coupled with mass spectrometry. Results were analysed using Student's paired t-test, at a 5% level of significance. Values of pharmacokinetic parameters that differed significantly between the two dosage regimens included the following: maximal concentration and area under the concentration-time curve were higher at 11 mg/kg, once daily, than at 5.5 mg/kg, twice daily; and, more importantly, the ratio of AUC(0-24) to the minimal inhibitory concentration (MIC) value of 0.5 µg/mL for a 24 h period (AUC(0-24)/MIC) was higher when clindamycin was administered at 11 than at 5.5 mg/kg, at least during the first day of drug administration. Therefore, a better pharmacokinetic profile may be expected when clindamycin is administered at 11 mg/kg, once daily, for the treatment of canine pyoderma caused by Staphylococcus pseudintermedius.