Antiviral therapy in cats progressively infected with feline leukaemia virus: lessons from a series of 18 consecutive cases from Australia.

BACKGROUND: It is doubtful that any of the treatments proposed for feline leukaemia virus (FeLV) infection are effective, despite the entity being described 60 years ago. METHODS: Eighteen pet cats with progressive FeLV infections were recruited in Australia. One or more antiviral drugs were trialled in 16 cats, while two FeLV-infected cats were not handleable and served as untreated controls. Six cats were administered RetroMAD1™ only (0.5 mg/kg orally twice daily), a commercially available recombinant chimeric protein with proposed antiretroviral activity. Three cats were administered the integrase inhibitor raltegravir only (10-15 mg/kg orally twice daily), a drug used as a component of highly effective antiretroviral therapy for human immunodeficiency virus (HIV-1) infection. Three cats were administered RetroMAD1™ and raltegravir concurrently, and four cats were administered raltegravir and the reverse transcriptase inhibitor zidovudine (AZT, 5 mg/kg orally twice daily) concurrently. FeLV RNA and p27 antigen loads were measured at two timepoints (T1-2 months and T3-5 months) during therapy and compared to baseline (pretreatment) levels, to assess the response to therapy using linear modelling. The median survival time (MST) of the cats from commencement of FeLV treatment to death was also determined and compared between treatments. RESULTS: The MST for the 16 FeLV-positive cats which received antiviral therapy was 634 days, while the MST from FeLV diagnosis to death for the two untreated control cats was 780 days. In cats treated with RetroMAD1™, FeLV viral load decreased from T0 to T1-2 months (median viral load reduced from 1339 × 106 to 705 × 106 copies/mL plasma; P = 0.012), but MST was reduced compared to cats not given RetroMAD1™ (426 days vs 1006 days; P = 0.049). Cats treated with raltegravir and AZT had no significant changes in FeLV viral load over time, but p27 antigen load was decreased from T0 to T3-5 months in cats treated with raltegravir (median p27 antigen level reduced from 50.2% to 42.7%; P = 0.005). All other results were not significantly affected by the treatment provided. Importantly, statistically significant and substantial associations were found between age at FeLV diagnosis and survival time (P = 0.046, R2 = 18.6) and between FeLV viral load at T0 and survival time (P = 0.004, R2 = 44.4). Younger cats, and cats with higher levels of pretreatment FeLV RNA, had reduced survival times. Cats treated with RetroMAD1™ were typically younger (median age 2.0 vs 8.0 years), likely explaining the observed reduction in MST. A significant association was found between FeLV viral load and p27 antigen load at T0 (P = 0.015, R2 = 32.9). CONCLUSIONS: Results from this small case series do not provide convincing support for the use of RetroMAD1™, raltegravir or AZT, alone or in combination, for the treatment of cats progressively infected with FeLV. The changes observed were biologically insignificant. Age and FeLV viral load at diagnosis are useful prognostic markers, and p27 antigen concentration can be used to predict viral load. Larger field trials should be performed examining antiretroviral therapy in FeLV-positive cats with progressive infections, preferably using three or more drugs from at least two classes, as is standard with human antiretroviral therapy. Future studies would be easier in countries with a higher prevalence of FeLV infections than Australia.

[SARS-CoV-2 infections in cats, dogs, and other animal species: Findings on infection and data from Switzerland]. / SARS-CoV-2 Infektionen bei Katzen, Hunden und anderen Tieren: Erkenntnisse zur Infektion und Daten aus der Schweiz.

INTRODUCTION: The pandemic with the novel coronavirus (SARS-CoV-2) has led to infections and deaths worldwide. Apart from humans, certain animal species are susceptible to the viral infection. Spillover between humans and animals is favored by close contact; thus, surveillance of animals is an important component to fight the pandemic from a One Health perspective. The Clinical Laboratory of the Vetsuisse Faculty Zurich has been investigating SARS-CoV-2 infections in animals since the beginning of the pandemic. In November 2020, the first SARS-CoV-2 positive Swiss cat was reported to the World Organisation for Animal Health (OIE-WAHIS). The cat showed respiratory signs and lived in a COVID-19 affected household. By now, over 500 natural SARS-CoV-2 infections have been recorded in animals worldwide. A prevalence study on SARS-CoV-2 infections in dogs and cats was carried out together with clinics from Germany and Italy during the first wave of the pandemic (March-July 2020). Among the tested 1137 animals, only one cat and one dog were positive. The prevalence of infection in dogs and cats presented to veterinary clinics was low, even in pandemic hotspot regions. However, recent studies that focused on animals in COVID-19 households found a higher prevalence of infection. A study is currently underway that specifically collects samples from pets from Swiss COVID-19 affected household and collects data on human-animal interaction.

INTRODUCTION: La pandémie à nouveau coronavirus (SARS-CoV-2) a entraîné des infections et des décès dans le monde entier. En dehors de l'homme, certaines espèces animales sont sensibles à cette infection virale. Le passage entre les humains et les animaux est favorisé par un contact étroit, la surveillance des animaux est donc un élément important pour lutter contre la pandémie dans une perspective One Health. Depuis le début de la pandémie, le laboratoire clinique de la faculté Vetsuisse de Zurich étudie les infections par le SRAS-CoV-2 chez les animaux. En novembre 2020, le premier chat suisse positif au SARS-CoV-2 a été signalé à l'Organisation mondiale de la santé animale (OIE-WAHIS). Le chat a montré des signes respiratoires et vivait dans un ménage touché par le COVID-19. À l'heure actuelle, plus de 500 infections naturelles au SRAS-CoV-2 ont été enregistrées chez des animaux dans le monde. Une étude de prévalence sur les infections par le SRAS-CoV-2 chez les chiens et les chats a été réalisée avec des cliniques d'Allemagne et d'Italie pendant la première vague de la pandémie (mars-juillet 2020). Parmi les 1137 animaux testés, seuls un chat et un chien étaient positifs. La prévalence de l'infection chez les chiens et les chats présentés aux cliniques vétérinaires était faible, même dans les régions fortement touchées par la pandémie. Cependant des études récentes, qui se sont concentrées sur les animaux dans les ménages COVID-19, ont révélé une prévalence d'infection plus élevée. Une étude est actuellement en cours qui collecte spécifiquement des échantillons d'animaux de compagnie des ménages suisses touchés par le COVID-19 et enregistre des données sur l'interaction homme-animal.

The bovine cumulus proteome is influenced by maturation condition and maturational competence of the oocyte.

In vitro maturation (IVM) of oocytes has still a negative impact on the developmental competence of oocytes. Therefore, this study analysed the cumulus proteome of individual cumulus-oocyte complexes (COCs) with and without maturational competence, matured under in vivo or in vitro conditions (n = 5 per group). A novel, ultrasensitive mass spectrometry (MS) based protein profiling approach, using label-free quantification, was applied. The detected cumulus proteome included 2226 quantifiable proteins and was highly influenced by the maturation condition (479 differentially expressed proteins) as well as maturational competence of the corresponding oocyte (424 differentially expressed proteins). Enrichment analysis showed an overrepresentation of the complement and coagulation cascades (CCC), ECM-receptor interaction and steroid biosynthesis in cumulus of COCs that matured successfully under in vivo conditions. Verification of the origin of CCC proteins was achieved through detection of C3 secretion into the maturation medium, with significantly increasing concentrations from 12 (48.4 ng/ml) to 24 hours (68 ng/ml: p < 0.001). In relation, concentrations in follicular fluid, reflecting the in vivo situation, were >100x higher. In summary, this study identified important pathways that are impaired in IVM cumulus, as well as potential markers of the maturational competence of oocytes.

Investigation of the Status Quo of Veterinary Point-of-Care Laboratories in Switzerland: Availability, Application, and Quality Management.

INTRODUCTION: The extent to which Swiss veterinary practitioners follow the guidelines for quality assurance of the American Society for Veterinary Clinical Pathology (ASVCP) for point-of-care (POC) testing is unknown. Thus, the aim of this study was to assess the availability, application, and quality management of POC analyzers in Swiss veterinary practices/clinics. For this purpose, we created an online questionnaire on laboratory equipment, quality management, and biosafety, which all members of the Society of Swiss Veterinarians (GST) were invited to complete. In total, 192 clinics/practices participated, of which 69% had automated POC analyzers, mainly for clinical chemistry (99%) and/or hematology (86%). Sample analyses and equipment maintenance were mostly performed by veterinary technicians (81% and 68%, respectively). Reference intervals were adopted from manufacturers (80%) or literature (17%). The results showed that most participants perform basic internal quality control (chemistry: 75%; hematology: 86%), and many use at least two levels of quality control material (47%-48%). Controls are mostly run once a month (chemistry: 36%; hematology: 35%) or ≤4 times/year (36% and 25%). Only three clinics/practices reported participation in an external quality assessment program; comparative testing was more common (chemistry: 42%; hematology: 52%). Only one-quarter of the participants stated that they make use of the data generated through internal and external quality control measures. In conclusion, POC analyzers are widely available in Swiss veterinary clinics/practices, and internal quality control is performed to some extent. However, quality assessment and management and biosafety awareness and measures need to be improved, ideally with the support of clinical pathologists.

INTRODUCTION: On ignore dans quelle mesure les vétérinaires suisses respectent les directives d'assurance qualité de l'American Society for Veterinary Clinical Pathology (ASVCP) pour les tests au point de service (Point of Care, POC). Ainsi, l'objectif de cette étude était d'évaluer la disponibilité, l'application et la gestion de la qualité des analyseurs POC dans les cabinets/cliniques vétérinaires suisses. À cette fin, nous avons créé un questionnaire en ligne sur les équipements de laboratoire, la gestion de la qualité et la biosécurité que tous les membres de la Société suisse des vétérinaires (GST) ont été invités à remplir. Au total, 192 cliniques/cabinets ont participé, dont 69% avaient des analyseurs POC automatisés, principalement pour la chimie clinique (99%) et/ou l'hématologie (86%). Les analyses des échantillons et la maintenance de l>équipement ont été principalement effectuées par des assistant(e)s en médecine vétérinaires (81% et 68%, respectivement). Les intervalles de référence ont été fixés sur la base des indications des fabricants (80%) ou de la littérature (17%). Les résultats ont montré que la plupart des participants effectuent un contrôle de qualité interne de base (chimie: 75%; hématologie: 86%) et que beaucoup utilisent au moins deux niveaux de matériel de contrôle de la qualité (47% ­48%). Les contrôles sont principalement effectués une fois par mois (chimie: 36%; hématologie: 35%) ou ≤4 fois / an (36% et 25%). Seules trois cliniques/cabinets ont déclaré avoir participé à un programme externe d'évaluation de la qualité. Les tests comparatifs étaient plus courants (chimie: 42%; hématologie: 52%). Un quart seulement des participants ont déclaré utiliser les données générées par des mesures de contrôle de qualité internes et externes. En conclusion, les analyseurs POC sont largement disponibles dans les cliniques/cabinets vétérinaires suisses et le contrôle qualité interne est effectué dans une certaine mesure. Cependant, l'évaluation et la gestion de la qualité ainsi que la sensibilisation et les mesures en matière de biosécurité doivent être améliorées, idéalement avec le soutien de pathologistes cliniciens.

Molecular investigations of cat fleas (Ctenocephalides felis) provide the first evidence of Rickettsia felis in Malta and Candidatus Rickettsia senegalensis in Israel.

Rickettsia felis, the causative agent of flea-borne spotted fever, occurs on all continents except Antarctica, owing to the cosmopolitan distribution of its cat flea vector. In this study, cat fleas were collected in two countries where the occurrence of R. felis was either unknown (Malta) or where accurate prevalence data were lacking (Israel). Altogether 129 fleas were molecularly analysed for the presence of rickettsial DNA. On the basis of three genetic markers, R. felis was identified in 39.5% (15/38) of the cat fleas from Malta. Sequences showed 100% identity to each other and to relevant sequences in GenBank. Among the 91 cat fleas from Israel, two (2.2%) contained the DNA of Candidatus Rickettsia senegalensis. Phylogenetically, the R. felis and Candidatus R. senegalensis identified here clustered separately (with high support) but within one clade, which was a sister group to that formed by the typhus group and spotted fever group rickettsiae. This is the first record of R. felis in Malta and of Candidatus R. senegalensis outside its formerly reported geographical range including Africa, Asia and North America.

[Feline leukemia virus infection: importance and current situation in Switzerland]. / Die feline Leukämievirus-Infektion: Bedeutung und aktuelle Situation in der Schweiz.

INTRODUCTION: Feline leukemia virus (FeLV) leads to fatal disease in cats with progressive infection. The aim of this study was to determine the importance of FeLV infection in Switzerland and make a comparison with previous studies. Of 881 blood samples taken from cats living in Switzerland (minimum of 20 samples per Canton), 47 samples were provirus-positive (5.3%; 95% confidence interval (CI) 3.9-7.0%) and 18 samples were antigen-positive (2%; 95% CI 1.2-3.2%). Together with data previously collected in similar studies, these findings demonstrated a decrease in prevalence between 1997 and 2003 followed by a relative constant low prevalence thereafter. Young cats (=2 years) were more frequently infected than older cats, but FeLV-positive cats were up to 15 (antigen-positive) and 19 (provirus-positive) years old. Sexually intact cats were more frequently viremic than neutered cats; purebred cats were somewhat less frequently FeLV-positive than non-purebred cats. In a second study, in which 300 saliva samples were analyzed, samples from 5 cats were FeLV-RNA positive (1.7%; 95% CI, 0.5-3.8%), although one young feral cat had been falsely assumed to be FeLV-negative based on a point-of-care test. Of the 300 cats, only 50% were FeLV tested or vaccinated, although 90% of the cats were at risk of exposure to FeLV. Testing and vaccination of all cats with exposure risk may help further decrease the prevalence of FeLV infection. Moreover, characteristics of FeLV tests should be considered, such as the risk of false negative results in the early phase of infection when performing antigen testing.

INTRODUCTION: Le virus leucémogène félin (FeLV) conduit la plupart du temps à une maladie mortelle chez le chat avec une infection progressive. Le but du présent travail est de mettre en évidence l'importance de l'infection à FeLV en Suisse sur la base de recherches actuelles et de la comparer avec les résultats de recherches antérieures. Afin de répondre à la question de savoir combien de chats présentés à la consultation étaient porteurs du FeLV (positifs au provirus) respectivement excréteurs de FeLV (positifs à l'antigène), on a analysé 881 échantillons sanguins provenant de toute la Suisse (au minimum 20 par canton) : 47 échantillons étaient positifs au provirus (5.3%; 95% intervalle de confiance (CI) 3.9­7.0%) et 18 positifs à l'antigène (2%; 95% CI 1.2­3.2%). Une comparaison avec des recherches semblables faites antérieurement montre que la prévalence du FeLV a diminué entre 1997 et 2003 mais qu'elle stagne depuis lors. Actuellement ce sont plutôt les jeunes chats (=2 ans) qui sont touchés plutôt que les vieux; des chats ont toutefois été trouvés positifs jusqu'à l'âge de 15 ans (positifs à l'antigène) respectivement de 19 ans (positifs au provirus). Les chats non castrés étaient plus souvent virémiques que les castrés et les chats de races étaient aussi, mais un peu moins fréquemment FeLV-positifs. Dans une autre étude suisse, dans laquelle 300 échantillons de salive de chats ont été testés quant à la présence d'ARN-FeLV, 5 chats étaient excréteurs (1.7%; 95% CI 0.5­3.8%). Un jeune chat trouvé, qui avait été testé négatif au test rapide, a été trouvé infecté par le FeLV au moyen de la mise en évidence d'ARN. Sur ces 300 chats, seuls environ 50% avaient été testés quant au FeLV respectivement vaccinés, bien qu'environ 90% aient présenté un risque d'exposition au FeLV. Pour diminuer encore la prévalence du FeLV, il conviendrait de tester et de vacciner tous les chats avec un risque d'exposition au virus. Dans ce contexte, il faut tenir compte des différentes caractéristiques des tests comme la non reconnaissance de la phase d'infection très précoce au moyen du test FeLV rapide.

Altered Serum Thyrotropin Concentrations in Dogs with Primary Hypoadrenocorticism before and during Treatment.

BACKGROUND: Thyrotropin (TSH) can be increased in humans with primary hypoadrenocorticism (HA) before glucocorticoid treatment. Increase in TSH is a typical finding of primary hypothyroidism and both diseases can occur concurrently (Schmidt's syndrome); therefore, care must be taken in assessing thyroid function in untreated human patients with HA. OBJECTIVE: Evaluate whether alterations in cTSH can be observed in dogs with HA in absence of primary hypothyroidism. ANIMALS: Thirty dogs with newly diagnosed HA, and 30 dogs in which HA was suspected but excluded based on a normal ACTH stimulation test (controls) were prospectively enrolled. METHODS: cTSH and T4 concentrations were determined in all dogs and at selected time points during treatment (prednisolone, fludrocortisone, or DOCP) in dogs with HA. RESULTS: cTSH concentrations ranged from 0.01 to 2.6 ng/mL (median 0.29) and were increased in 11/30 dogs with HA; values in controls were all within the reference interval (range: 0.01-0.2 ng/dL; median 0.06). There was no difference in T4 between dogs with increased cTSH (T4 range 1.0-2.1; median 1.3 µg/dL) compared to those with normal cTSH (T4 range 0.5-3.4, median 1.4 µg/dL; P=0.69) and controls (T4 range 0.3-3.8, median 1.8 µg/dL; P=0.35). After starting treatment, cTSH normalized after 2-4 weeks in 9 dogs and after 3 and 4 months in 2 without thyroxine supplementation. CONCLUSIONS AND CLINICAL RELEVANCE: Evaluation of thyroid function in untreated dogs with HA can lead to misdiagnosis of hypothyroidism; treatment with glucocorticoids for up to 4 months can be necessary to normalize cTSH.

Detection of 'Candidatus Neoehrlichia mikurensis' and other Anaplasmataceae and Rickettsiaceae in Canidae in Switzerland and Mediterranean countries.

INTRODUCTION: 'Candidatus Neoehrlichia mikurensis' is an emerging tick-borne zoonotic agent that primarily affects immunocompromised human patients. Dogs and foxes are frequently exposed to ticks, and both species are in close proximity to humans. This is the first study to systematically investigate the occurrence of 'Candidatus Neoehrlichia mikurensis' in Canidae in Europa. We analyzed 1'739 blood samples from dogs in Switzerland, Italy, Spain and Portugal and 162 blood samples from free-ranging red foxes (Vulpes vulpes) in Switzerland. All samples were tested using a previously described multiplex real-time PCR for the Anaplasmataceae family, the 'Candidatus Neoehrlichia' genus and the 'Candidatus Neoehrlichia mikurensis' species. All Anaplasmataceae positive samples were subsequently tested using specific real-time PCRs for Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis and Rickettsia helvetica. Among the tested animals, one dog from Zurich tested positive for 'Candidatus Neoehrlichia mikurensis'. The 12-year old West Highland white terrier had been splenectomized 3 months prior to the blood collection and presented with polyuria/polydipsia. Fanconi syndrome was diagnosed based on glucosuria with normoglycemia and hyperaminoaciduria. A. platys and E. canis were detected in 14/249 dogs from Sicily and Portugal; two of the dogs were coinfected with both agents. Four Swiss foxes tested positive for A. phagocytophilium. R. helvetica was detected for the first time in a red fox. In conclusion, 'Candidatus Neoehrlichia mikurensis' infection should be considered in sick dogs, particularly when immunocompromised. The pathogen seems not to be widespread in Canidae in the investigated countries. Conversely, other Anaplasmataceae were more readily detected in dogs and foxes.

INTRODUCTION: 'Candidatus Neoehrlichia mikurensis' est un agent de zoonose transmis par les tiques qui gagne en importance et concerne principalement les patients immunosupprimés. Les chiens comme les renards sont souvent concernés par des morsures de tiques et vivent en contact étroit avec les êtres humains. Dans le présent travail, nous étudions pour la première fois systématiquement la présence de 'Candidatus Neoehrlichia mikurensis' chez les canidés en Europe. Les échantillons sanguins analysés provenaient de 1'739 chiens de Suisse, d'Italie, d'Espagne et du Portugal ainsi que de 162 renards (Vulpes vulpes) de Suisse. Tous les échantillons ont été examinés avec un test de PCR multiplex en temps réel déjà publié quant à la présence d'agents de la famille des Anaplasmataceae, du genre 'Candidatus Neoehrlichia' et de l'espèce 'Candidatus Neoehrlichia mikurensis'. Les échantillons positifs aux Anaplasmataceae ont ensuite été testés avec un test PCR en temps réel spécifique quant à Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis und Rickettsia helvetica. Parmi les échantillons examinés se trouvait celui d'un chien de Zürich qui était infecté par 'Candidatus Neoehrlichia mikurensis'. Ce West Highland White Terrier de 12 ans avait été présenté pour polyurie/polydipsie; il avait été splénectomisé trois mois avant la prise de l'échantillon. Au vu d'une glycosurie et d'une hyperaminoacidurie accompagnées d'une glycémie normale, on a posé le diagnostic de syndrome de Fanconi. A. platys et E. canis ont été mis en évidence chez 14/249 chiens provenant de Sicile et du Portugal; deux chiens étaient infectés par les deux agents pathogènes. Quatre renards suisses étaient positifs à A. phagocytophilium et R. helvetica a été trouvé pour la première fois chez un renard. En résumé, on peut dire qu'une infection à 'Candidatus Neoehrlichia mikurensis' chez un chien malade doit être prise en considération comme diagnostic différentiel, particulièrement chez les anomaux immunosupprimés. Toutefois cet agent n'est pas très répandu chez les canidés des pays examinés, contrairement aux autres Anaplasmataceae spp. qui ont été trouvées plus souvent chez les chiens et les renards.

Prognostic Markers in Acute Babesia canis Infections.

BACKGROUND: Canine babesiosis, caused by Babesia canis, is a prevalent and clinically relevant disease in Europe. Severe acute babesiosis is characterized by a high mortality but prognosis is not always correlated with clinical signs nor with the level of parasitemia. OBJECTIVE: This study evaluated prognostic markers associated with poor outcomes in acute Babesia canis infections. ANIMALS AND METHODS: We compared the results of routine laboratory profiles, hand-held lactate and glucose analyzer, and the acute phase response in 2 groups of naturally infected dogs (7 survivors and 8 nonsurvivors). Samples were collected at the time of first admission and before any treatment. Subsequently, the course of prognostic markers was followed in 3 dogs experimentally inoculated with B. canis. RESULTS: Nonsurvivors showed significantly higher concentrations of lactate, triglycerides and phosphate and lower hematocrit, leukocyte counts, total serum protein concentrations, and thrombocyte counts when compared to survivors. All nonsurvivors (8/8) had hyperlactatemia, whereas most survivors (6/7) had values within the reference range. All survivors had leucocyte counts within the reference range, unlike the nonsurvivors, which showed leukopenia. During the course of acute babesiosis, the variables serum lactate, triglyceride, and phosphate concentrations, and thrombocyte count only exceeded a prognostic threshold during acute crisis. CONCLUSIONS AND CLINICAL IMPORTANCE: Poor outcome in acute B. canis infection is indicated by changes in the laboratory profile. Intensive care should be considered for dogs presenting with moderate anemia, severe thrombocytopenia, mild to moderate leukopenia, hyperlactatemia, moderately increased serum phosphate, and triglyceride concentrations, and moderately decreased total serum protein concentrations.

Cortisol Response in Healthy and Diseased Dogs after Stimulation with a Depot Formulation of Synthetic ACTH.

BACKGROUND: The ACTH stimulation test is used to evaluate the adrenocortical reserve. Recently, the availability of the synthetic ACTH formulation was limited, causing major problems in clinical practice. OBJECTIVES: The objective of this study was to evaluate poststimulation peak cortisol concentrations and the duration of the stimulatory effect of a depot ACTH preparation in dogs. ANIMALS: Twenty-two healthy dogs, 10 dogs with suspected hypoadrenocorticism (HA) and 15 dogs with suspected hyperadrenocorticism (HC). METHODS: Prospective study. An ACTH stimulation test using a synthetic depot tetracosactide, administered intramuscularly (5 µg/kg or at least 0.1 mL) was performed. Blood samples for determination of cortisol were taken immediately before and 1, 2, 3, 4, 6, and 24 hours after stimulation. RESULTS: Peak cortisol concentrations were reached after 2-4 hours in all dogs. Cortisol concentrations 1 hour after stimulation were >9 µg/dL in all healthy dogs and >5 µg/dL in all dogs in which HA was excluded. None of the dogs with HA showed a cortisol-increase above the detection-limit of the assay. After 6 hours, cortisol concentrations had decreased in the healthy and HC group and were back to baseline after 24 hours. CONCLUSIONS AND CLINICAL IMPORTANCE: The depot formulation can be used in place of the short-acting ACTH to evaluate the adrenocortical reserve. Blood for peak cortisol concentrations should be drawn 3 hours after stimulation in cases in which HC is suspected; in HA-suspected cases, blood sampling can take place after 1 hour. As the stimulatory effect is gone after 24 hours, interference with other hormonal tests is unlikely after that time.

Evaluation of the Cortisol-to-ACTH Ratio in Dogs with Hypoadrenocorticism, Dogs with Diseases Mimicking Hypoadrenocorticism and in Healthy Dogs.

BACKGROUND: The adrenocorticotropic hormone (ACTH) stimulation test is the gold standard for diagnosing hypoadrenocorticism (HA) in dogs. However, problems with the availability of synthetic ACTH (tetracosactrin/cosyntropin) and increased costs have prompted the need for alternative methods. OBJECTIVES: To prospectively evaluate the cortisol-to-ACTH ratio (CAR) as a screening test for diagnosing canine HA. ANIMALS: Twenty three dogs with newly diagnosed HA; 79 dogs with diseases mimicking HA; 30 healthy dogs. METHODS: Plasma ACTH and baseline cortisol concentrations were measured before i.v. administration of 5 µg/kg ACTH in all dogs. CAR was calculated and the diagnostic performance of ACTH, baseline cortisol, CAR and sodium-to-potassium ratios (SPRs) was assessed based on receiver operating characteristics (ROC) curves calculating the area under the ROC curve. RESULTS: The CAR was significantly lower in dogs with HA compared to that in healthy dogs and in those with diseases mimicking HA (P < .0001). There was an overlap between HA dogs and those with HA mimicking diseases, but CAR still was the best parameter for diagnosing HA (ROC AUC 0.998), followed by the ACTH concentration (ROC AUC 0.97), baseline cortisol concentration (ROC AUC 0.96), and SPR (ROC AUC 0.86). With a CAR of >0.01 the diagnostic sensitivity and specificity were 100% and 99%, respectively. CONCLUSION AND CLINICAL IMPORTANCE: Calculation of the CAR is a useful screening test for diagnosing primary HA. As a consequence of the observed overlap between the groups, however, misdiagnosis cannot be completely excluded. Moreover, additional studies are needed to evaluate the diagnostic reliability of CAR in more dogs with secondary HA.

Urinary and plasma catecholamines and metanephrines in dogs with pheochromocytoma, hypercortisolism, nonadrenal disease and in healthy dogs.

BACKGROUND: Diagnosis of pheochromocytoma (PC) is based on a combination of clinical suspicion, finding an adrenal mass, increased plasma, and urine concentrations of catecholamine metabolites and is finally confirmed with histopathology. In human medicine, it is controversial whether biochemically testing plasma is superior to testing urine. OBJECTIVES: To measure urinary and plasma catecholamines and metanephrines in healthy dogs, dogs with PC, hypercortisolism (HC), and nonadrenal diseases (NAD) and to determine the test with the best diagnostic performance for dogs with PC. ANIMALS: Seven PC dogs, 10 dogs with HC, 14 dogs with NAD, 10 healthy dogs. METHODS: Prospective diagnostic clinical study. Urine and heparin plasma samples were collected and stored at -80°C before analysis using high-pressure liquid chromatography (HPLC) coupled to electrochemical detection or tandem mass spectrometry were performed. Urinary variables were expressed as ratios to urinary creatinine concentration. RESULTS: Dogs with PC had significantly higher urinary normetanephrine and metanephrine:creatinine ratios and significantly higher plasma-total and free normetanephrine and plasma-free metanephrine concentrations compared to the 3 other groups. There were no overlapping results of urinary normetanephrine concentrations between PC and all other groups, and only one PC dog with a plasma normetanephrine concentration in the range of the dogs with HC and NAD disease. Performances of total and free plasma variables were similar. Overlap of epinephrine and norepinephrine results between the groups was large with both urine and plasma. CONCLUSION AND CLINICAL IMPORTANCE: Measurement of normetanephrine is the preferred biochemical test for PC and urine was superior to plasma.

Urinary corticoid concentrations measured by 5 different immunoassays and gas chromatography-mass spectrometry in healthy dogs and dogs with hypercortisolism at home and in the hospital.

BACKGROUND: Determination of the urinary corticoid-to-creatinine ratio (UCCR) is an important screening test in the diagnosis of hypercortisolism (HC). However, urinary cortisol metabolites interfere with cortisol measurement in immunoassays, leading to decreased specificity. Gas chromatography-mass spectrometry (GC-MS) is considered the gold standard for steroid hormone analysis, because it provides a high level of selectivity and accuracy. OBJECTIVES: To prospectively compare the UCCR of healthy dogs and dogs with HC determined by 5 different immunoassays and by GC-MS and to evaluate the influence of veterinary care on UCCR. ANIMALS: Twenty healthy dogs; 18 dogs with HC. METHODS: Urine was collected in the hospital and again after 6 days at home. Three chemiluminescence immunoassays (Access 2, Beckmann; Immulite 2000, DPC Siemens, with and without trichloromethane extraction) and 2 RIAs (Utrecht in house; Access Beckmann) were used. GC-MS analyses were performed with Agilent 6890N/5973N. Urinary corticoid concentrations were related to urinary creatinine concentrations. RESULTS: Immunoassay results were significantly higher compared to GC-MS results. Evaluation of bias plots and clinical assessment made on the basis of the assay results of each dog indicated substantial disagreement among the assays. Sensitivity varied from 37.5 to 75% and with selected assays was lower in samples from day 6 compared to day 0. GC-MS was not superior to the immunoassays in discriminating healthy from HC dogs. CONCLUSIONS AND CLINICAL IMPORTANCE: Considerable variation must be anticipated comparing different urinary cortisol assays. Establishing an assay- and laboratory-specific reference range is critical when using UCCR.

Total protein measurement in canine cerebrospinal fluid: agreement between a turbidimetric assay and 2 dye-binding methods and determination of reference intervals using an indirect a posteriori method.

BACKGROUND: In veterinary clinical laboratories, qualitative tests for total protein measurement in canine cerebrospinal fluid (CSF) have been replaced by quantitative methods, which can be divided into dye-binding assays and turbidimetric methods. There is a lack of validation data and reference intervals (RIs) for these assays. OBJECTIVES: The aim of the present study was to assess agreement between the turbidimetric benzethonium chloride method and 2 dye-binding methods (Pyrogallol Red-Molybdate method [PRM], Coomassie Brilliant Blue [CBB] technique) for measurement of total protein concentration in canine CSF. Furthermore, RIs were determined for all 3 methods using an indirect a posteriori method. METHODS: For assay comparison, a total of 118 canine CSF specimens were analyzed. For RIs calculation, clinical records of 401 canine patients with normal CSF analysis were studied and classified according to their final diagnosis in pathologic and nonpathologic values. RESULTS: The turbidimetric assay showed excellent agreement with the PRM assay (mean bias 0.003 g/L [-0.26-0.27]). The CBB method generally showed higher total protein values than the turbidimetric assay and the PRM assay (mean bias -0.14 g/L for turbidimetric and PRM assay). From 90 of 401 canine patients, nonparametric reference intervals (2.5%, 97.5% quantile) were calculated (turbidimetric assay and PRM method: 0.08-0.35 g/L (90% CI: 0.07-0.08/0.33-0.39); CBB method: 0.17-0.55 g/L (90% CI: 0.16-0.18/0.52-0.61). Total protein concentration in canine CSF specimens remained stable for up to 6 months of storage at -80°C. CONCLUSIONS: Due to variations among methods, RIs for total protein concentration in canine CSF have to be calculated for each method. The a posteriori method of RIs calculation described here should encourage other veterinary laboratories to establish RIs that are laboratory-specific.

Duration of t4 suppression in hyperthyroid cats treated once and twice daily with transdermal methimazole.

BACKGROUND: Transdermal methimazole is an acceptable alternative to oral treatment for hyperthyroid cats. There are, however, no studies evaluating the duration of T4 suppression after transdermal methimazole application. Such information would be valuable for therapeutic monitoring. OBJECTIVE: To assess variation in serum T4 concentration in hyperthyroid cats after once- and twice-daily transdermal methimazole administration. ANIMALS: Twenty client-owned cats with newly diagnosed hyperthyroidism. METHODS: Methimazole was formulated in a pluronic lecithin organogel-based vehicle and applied to the pinna of the inner ear at a starting dose of 2.5 mg/cat q12h (BID group, 10 cats) and 5 mg/cat q24h (SID group, 10 cats). One and 3 weeks after starting treatment, T4 concentrations were measured immediately before and every 2 hours after gel application over a period of up to 10 hours. RESULTS: Significantly decreased T4 concentrations were observed in week 1 and 3 compared with pretreatment concentrations in both groups. All cats showed sustained suppression of T4 concentration during the 10-hour period, and T4 concentrations immediately before the next methimazole treatment were not significantly different compared with any time point after application, either in the BID or SID groups. CONCLUSIONS: Because transdermal methimazole application led to prolonged T4 suppression in both the BID and SID groups, timing of blood sampling does not seem to be critical when assessing treatment response.

[Haemoplasma infection in a dairy cow]. / Hämoplasmeninfektion bei einer Milchkuh.

The present work describes the clinical and laboratory examination as well as the treatment of a 7-year-old local dairy breed cow presented with reduced appetite, decreasing milk yield and striking yellowish discoloured skin and mucosa. The laboratory examination revealed a high degree regenerative anaemia and hyperbilirubinaemia. The bovine haemotrophic mycoplasma species Mycoplasma wenyonii and 'Candidatus Mycoplasma haemobos' were detected in the blood by PCR. Treatment with oxytetracycline rapidly improved the general condition, and milk production was increased. In a follow-up study, blood samples of all 23 animals from the same herd were examined. Fifteen cows were found to be infected with both haemoplasma species, three animals were only infected with 'Candidatus Mycoplasma haemobos' and one animal only with Mycoplasma wenyonii. Two out of three tested calves were positive for 'Candidatus Mycoplasma haemobos'. Except for the above described anaemic cow, all other animals were clinically healthy.

[Primary hyperaldosteronism in cats]. / Primärer Hyperaldosteronismus bei Katzen.

Primary hyperaldosteronism is a clinical syndrome characterized by an elevated aldosterone secretion by the adrenals. The present case series describes 7 cats with primary hyperaldosteronism, which were presented between 2002 and 2011. Common clinical symptoms were weakness, anorexia, cervical ventroflexion and blindness. All cats showed hypokalemia. In 6 cats, blood pressure was determined: 5 cats showed hypertension, of which 4 animals exhibited retinal detachment and blindness. In the ultrasonographic examination, unilateral adrenomegaly was present in 6 cats whereas one animal showed normal adrenals. In 4 cats, the serum aldosterone concentration was above the reference range. Five cats underwent unilateral adrenalectomy, which was accomplished uneventfully and returned the electrolytes back to normal. Histopathological examination of the adrenals revealed 2 carcinomas and 4 adenomas; one cat with ultrasonographic normal adrenals exhibited bilateral nodular hyperplasia.

Fatal bovine anaplasmosis in a herd with new genotypes of Anaplasma marginale, Anaplasma ovis and concurrent haemoplasmosis.

Haematological and molecular analysis of blood samples was carried out during an outbreak of bovine anaplasmosis in Hungary. Acute disease was observed in five animals, two of which died. Anaplasma-carrier state was diagnosed in 69 (92%) of cattle. Further evaluation of 24 blood samples revealed concurrent infections with Mycoplasma wenyonii and 'CandidatusM. haemobos' in 22 and 21 animals, respectively. In addition, two cows were identified with rickettsaemia. Regarding molecular investigation of potential hard tick vectors, Haemaphysalis inermis and Dermacentor marginatus males collected from the animals were PCR-negative. However, in one pool (out of 18) of Ixodesricinus males, and in six pools (out of 18) of D. reticulatus males the msp4 gene of Anaplasma marginale was detected. In the same I. ricinus pool Anaplasma ovis was also identified. All ticks were negative for haemoplasmas. Anaplasma sequences yielded 97-99% homology to sequences deposited in the Genbank. This is the first report of fatal bovine anaplasmosis associated with divergent A. marginale genotypes and concurrent 'CandidatusM. haemobos' infection, as well as of an A. ovis strain in ticks collected from cattle.

[FeLV infection in the cat: clinically relevant aspects]. / Die FeLV-Infektion der Katze: Praxis-relevante Aspekte.

The feline leukemia virus (FeLV) is a retrovirus of the domestic cat that was described almost 50 years ago. The FeLV-infection may lead to fatal diseases in domestic and small wild cats. The use of efficacious diagnostics assays and vaccines led to a reduction of the FeLV prevalence; however, FeLV still poses a problem for the cat presented with the infection. This article aims to describe recent developments in diagnostics and findings in the infection pathogenesis that are clinically relevant.