[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.

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.

[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.

Feline leukemia virus outbreak in the critically endangered Iberian lynx (Lynx pardinus): high-throughput sequencing of envelope variable region A and experimental transmission.

The Iberian lynx is the most endangered felid species. During winter/spring 2006/7, a feline leukemia virus (FeLV) outbreak of unexpected virulence killed about 2/3 of the infected Iberian lynxes. All FeLV-positive animals were co-infected with feline hemoplasmas. To further characterize the Iberian lynx FeLV strain and evaluate its potential virulence, the FeLV envelope gene variable region A (VRA) mutant spectrum was analyzed using the Roche 454 sequencing technology, and an in vivo transmission study of lynx blood to specified-pathogen-free cats was performed. VRA mutations indicated weak apolipoprotein B mRNA editing enzyme and catalytic polypeptide-like cytidine deaminase (APOBEC) restriction of FeLV replication, and variants characteristic of aggressive FeLV strains, such as FeLV-C or FeLV-A/61C, were not detected. Cats exposed to FeLV/Candidatus Mycoplasma haemominutum-positive lynx blood did not show a particularly severe outcome of infection. The results underscore the special susceptibility of Iberian lynxes to infectious diseases.

Human neutralizing monoclonal antibodies of the IgG1 subtype protect against mucosal simian-human immunodeficiency virus infection.

Although maternal human immunodeficiency virus type 1 (HIV-1) transmission occurs during gestation, intrapartum and postpartum (by breast-feeding), 50-70% of all infected children seem to acquire HIV-1 shortly before or during delivery. Epidemiological evidence indicates that mucosal exposure is an important aspect of intrapartum HIV transmission. A simian immunodeficiency virus (SIV) macaque model has been developed that mimics the mucosal exposure that can occur during intrapartum HIV-1 transmission. To develop immunoprophylaxis against intrapartum HIV-1 transmission, we used SHIV-vpu+ (refs. 5,6), a chimeric simian-human virus that encodes the env gene of HIV-IIIB. Several combinations of human monoclonal antibodies against HIV-1 have been identified that neutralize SHIV-vpu+ completely in vitro through synergistic interaction. Here, we treated four pregnant macaques with a triple combination of the human IgG1 monoclonal antibodies F105, 2G12 and 2F5. All four macaques were protected against intravenous SHIV-vpu+ challenge after delivery. The infants received monoclonal antibodies after birth and were challenged orally with SHIV-vpu+ shortly thereafter. We found no evidence of infection in any infant during 6 months of follow-up. This demonstrates that IgG1 monoclonal antibodies protect against mucosal lentivirus challenge in neonates. We conclude that epitopes recognized by the three monoclonal antibodies are important determinants for achieving substantial protection, thus providing a rational basis for AIDS vaccine development.

[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.

Prevalence of dog erythrocyte antigen 1.1 in dogs in Switzerland evaluated with the gel column technique.

Canine blood typing has become an established and essential laboratory test due to the rising demand for safe and efficient blood transfusions. The most immunogenic and clinically important blood type is DEA 1.1. Little is known about DEA 1.1 frequencies or special characteristics among different canine breeds. 304 dogs were tested for DEA 1.1. DEA 1.1-typing was performed using a commercial gel column technique (ID-Gel Test Canine DEA 1.1, DiaMed, Cressier, Switzerland). Fifty-three percent of all tested dogs reacted positive for DEA 1.1, whereas 49 % of the mixed breeds tested DEA 1.1-positive. All Bernese mountain dogs (n = 22) and Rottweilers (n = 9) tested positive for DEA 1.1, while all Boxers (n = 8), Flat-Coated Retrievers (n = 9), and Border Collies (6) tested negative for DEA 1.1. The prevalence of DEA 1.1 in dogs in Switzerland was found to be comparable to that reported from other countries. The tested breeds were found to differ considerably in the frequency of DEA 1.1. This knowledge is useful for selection of blood donors. However, DEA 1.1 blood typing of donor and recipient prior to transfusion and cross matching in sensitized dogs is unavoidable.

Housing and care of laboratory cats: from requirements to practice.

Increased public awareness of the welfare and well-being of laboratory animals in biomedical research and related ethical considerations inspired us to review recent developments and recommendations for the care and housing of laboratory cats. The present review focuses on the practical requirements for maintaining domestic cats as laboratory animals - from the construction of animal shelters to the termination of an experiment. An excellent standard of housing and care will reduce the bias of experimental results due to stress. To provide cats with living conditions that best meet their natural physical requirements and permit natural social behaviour, laboratories should spare no effort to achieve high housing standards. Hence, the present report not only aims to be a practical reference for those who are involved in the care and husbandry of cats, but it also aims to motivate researchers to improve their knowledge in this field and to provide humane conditions for all cats kept for scientific purposes.

[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.

Haemotropic mycoplasmas of cats and dogs: transmission, diagnosis, prevalence and importance in Europe.

Haemotropic mycoplasmas (or haemoplasmas) are the causative agents of infectious anaemia in many mammalian species. They were previously known as Haemobartonella and Eperythrozoon species. The development of sensitive, specific PCR assays has expanded our knowledge of these agents and PCR is the method of choice to diagnose and differentiate haemoplasma infections. In felids, Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum' and 'Candidatus Mycoplasma turicensis' have been described. They vary strongly in their pathogenic potential and co-factors may influence the disease severity. In dogs, Mycoplasma haemocanis and 'Candidatus Mycoplasma haematoparvum' are known; clinical signs are mainly found in immunocompromised dogs. Transmission of haemoplasmas may occur via infected blood (aggressive interaction, transfusion) or blood-sucking arthropods. Infections can be treated with Doxycycline, although it is disputable whether the infection is completely eliminated. Feline haemoplasmas must be expected in cats all over Europe, while canine haemoplasmas are mainly encountered in dogs in Mediterranean countries but should also be considered in Swiss dogs with a travel history.

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.

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.

Comparison of 2 doses of recombinant human thyrotropin for thyroid function testing in healthy and suspected hypothyroid dogs.

BACKGROUND: Various protocols using different doses of recombinant human thyrotropin (rhTSH) in TSH stimulation testing have been described. However, the influence of TSH dosage on thyroxine (T4) concentration has not yet been evaluated in suspected hypothyroid dogs. OBJECTIVE: To evaluate the effectiveness of 2 doses of rhTSH. ANIMALS: Fifteen dogs with clinical signs consistent with hypothyroidism and abnormal stimulation results with 75 microg rhTSH and 18 clinically healthy dogs. METHODS: All dogs were stimulated with 75 and 150 microg rhTSH IV in a 1st and 2nd stimulation test, respectively. Blood samples were taken before and 6 hours after rhTSH administration for determination of total T4 concentration. RESULTS: Using the higher dose led to a normal test interpretation in 9 of the 15 dogs, in which stimulation had been abnormal using the lower dose. Based on follow-up information, hypothyroidism was excluded in 7 of these 9 dogs. In all 6 dogs with a blunted response to the higher dose, hypothyroidism could be confirmed. Healthy dogs showed significantly higher post-TSH T4 concentrations with the higher compared with the lower dose. Post-TSH T4 concentrations after TSH stimulation were not related to dogs' body weight in either healthy or diseased dogs. CONCLUSIONS AND CLINICAL RELEVANCE: TSH dose significantly influenced test interpretation in suspected hypothyroid dogs. Differentiation between primary hypothyroidism and nonthyroidal disease was improved with 150 microg rhTSH. Because this effect was independent of the dogs' body weight, the higher dose is recommended in dogs that have concurrent disease or are receiving medication.

Liquid culture medium for the rapid cultivation of Helicobacter pylori from biopsy specimens.

The goal of this study was to develop a liquid culture medium for the rapid isolation, cultivation, identification and subsequent antibiotics susceptibility testing of Helicobacter pylori directly from biopsy specimens. Five liquid media were tested: Ham's F-12, Brucella broth, tryptic soybroth, brain heart infusion broth and Mueller-Hinton broth. After optimisation of the medium, it was applied in order to investigate biopsy samples from 150 patients with gastro-duodenal disorders and compared with traditional culture methods, microscopy and an H. pylori-specific TaqMan real-time polymerase chain reaction (PCR). The most reliable and rapid growth of H. pylori, even at a small inoculum size, was obtained in Ham's F-12 medium with 5% horse serum. The developed system allowed the primary isolation of H. pylori in clinical samples and provided 87% sensitivity and 100% specificity.

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.

Genotyping of Babesia bigemina from cattle from a non-endemic area (Switzerland).

In August 2002, bovine anaplasmosis and concurrent infections with Mycoplasma sp. and piroplasms were reported in a cattle herd in an alpine region of Switzerland. The piroplasms were identified by PCR/sequencing of part of the 18S rRNA gene as Babesia bigemina and Theileria of the buffeli/sergenti/orientalis-complex, which have never been diagnosed in Switzerland before. The B. bigemina isolate was genetically characterised at two loci and compared with isolates from Italy, Spain, Turkey, Kenya and Mexico. Analysis of the internal transcribed spacer 2 (ITS2) of the rRNA genes revealed high polymorphism not only among the isolates but even within the isolates, and the presence of two types of the ITS2 in every isolate was confirmed. A dendrogram based on ITS2 sequences showed that the Swiss isolate was most closely related to a Spanish isolate but no sequences of the isolate from Switzerland were identical to any of the other isolates. The isolate from Italy was not positioned in the same cluster as the Swiss and the Spanish isolate. This had been anticipated as the nearest known endemic area of B. bigemina in Central Italy. Sequence analysis of the rhoptry-associated protein-1c gene (rap1c) confirmed the similarity of the Swiss and Spanish isolate. Hence, our molecular analyses of the Swiss B. bigemina isolate did not unequivocally track its geographical origin and the way of introduction remains obscure.

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.

Shedding of feline leukemia virus RNA in saliva is a consistent feature in viremic cats.

The purpose of this investigation was to characterize the shedding pattern of feline leukemia virus (FeLV) RNA in saliva, and to correlate it with the proviral load in whole blood, viral load in plasma, levels of p27 in saliva and plasma, the isolation of infectious FeLV from saliva, and the titers of FeLV-specific antibodies of the IgG and IgA isotypes. We evaluated 24 experimentally FeLV-infected cats for these parameters using real-time RT-PCR and PCR, cell culture assay and sandwich ELISA. We observed that shedding of viral RNA in saliva was a consistent feature in viremic cats. Latently FeLV-infected cats, displaying a very low proviral load, did not shed infectious virus in saliva, but occasionally shed viral RNA. Consequently, salivary shedding of FeLV RNA may not necessarily indicate a transmission potential for susceptible cats. This study also confirmed previous results from our laboratory, showing that a negative result for p27 in plasma, or for viral RNA in plasma or saliva does not exclude FeLV infection, considering that blood cells from those cats contained provirus. We also showed that FeLV RNA and DNA were stable for more than 64 days in saliva samples stored at room temperature. We conclude that the detection of FeLV RNA in saliva may be a useful indicator of viremia, and that the detection of salivary viral RNA by RT-PCR could become a reliable tool for the diagnosis of FeLV infection, which is facilitated by the low invasive method of collection of the samples.

[Feline hemoplasmas in Switzerland: identification of a novel species, diagnosis, prevalence, and clinical importance]. / Feline Haemoplasmen in der Schweiz: Identifikation einer neuen Spezies, Diagnose, Prävalenz und klinische Bedeutung.

Two feline hemotropic mycoplasma spp. (aka hemoplasma) have previously been recognized. We recently discovered a third novel species in a cat with hemolytic anemia, designated 'Candidatus Mycoplasma turicensis', which is closely related to rodent haemoplasmas. This novel species induced anemia after experimental transmission to two SPF cats. Three quantitative real-time PCR assays were newly designed and applied to an epidemiological study surveying the Swiss pet cat population. Blood samples from 713 healthy and ill cats were analyzed. Up to 104 parameters per cat (detailed questionnaire, case history, laboratory parameters and retroviral infections) were evaluated. 'Candidatus Mycoplasma haemominutum' infection was more prevalent (8.5%) than Mycoplasma haemofelis (0.5%) and 'Candidatus Mycoplasma turicensis' (1%). Hemoplasma infections were associated with male gender, outdoor access, and old age, but not with disease or anemia. Infections were more frequently found in the South and West of Switzerland. Several hemoplasma infected cats, some acutely infected, others co-infected with FIV or FeLV, showed hemolytic anemia indicating that additional factors might play a role in the pathogenesis of the disease.

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.