High-throughput sequencing of Strongyloides stercoralis - a fatal disseminated infection in a dog.

The rhabditid nematode Strongyloides stercoralis is known worldwide as the causative agent of strongyloidiasis in humans. In addition to public health concerns, S. stercoralis also infects dogs, which represent a possible reservoir for potentially zoonotic transmissions. We describe the first confirmed case of fatal disseminated infection in a dog in the Czech Republic. The microscopic and histological results were supported by a complex genotyping approach. Using high-throughput sequencing of the hypervariable region (HVR-IV) of 18S rDNA and Sanger sequencing of the partial cytochrome c oxidase subunit 1 gene (cox1), the potentially zoonotic haplotype/lineage A of S. stercoralis was confirmed, while the solely canine haplotype/lineage B was not found. The development of the disease is mainly associated with immunodeficiency, and in this case, it was triggered by inappropriate treatment, in particular the use of corticosteroids.

Canine leishmaniosis in three consecutive generations of dogs in Czech Republic.

Transmission of canine leishmaniosis (CanL) is described in three consecutive generations of female Boxers living in a non-endemic environment in the Czech Republic. Infection of the first generation female likely occurred during a breeding visit to Italy and the dog died with typical clinical signs of the disease but without definitive laboratory diagnosis. The second and third generation offsprings never left the Czech Republic, suffered from clinical CanL confirmed by polymerase chain reaction and serology, and were apparently infected by transplacental transmission. Persistence of CanL in the Czech Republic over 7 years with a suspected origin in an endemic region and progression of infection through subsequent generations in a non-endemic country exemplifies that this disease may establish itself also in areas where no obvious vectors are present.

InPouch™ TF-Feline medium is not specific for Tritrichomonas foetus.

Trichomonads are a group of anaerobic flagellates. Two species of intestinal trichomonads have been described in cats - pathogenic Tritrichomonas foetus and probably harmless Pentatrichomonas hominis. InPouch™ TF-Feline medium (Bio-Med Diagnostics, White City, Oregon, USA) is considered the gold standard for diagnosis of T. foetus infection in cats. It is commercially available, relatively cheap and easy-to-use. The medium is intended to be highly specific to T. foetus as morphologically similar Pentatrichomonas hominis and Giardia sp. do not survive here longer than 24h. In this study we successfully cultivated P. hominis in InPouch™ TF-Feline medium for 3 days after inoculation with cat faeces. The identity of the organism was assessed by sequencing of SSU rDNA and ITS region. Possible coinfection with T. foetus was ruled out using Tritrichomonas-specific PCR. Our results suggest possible misdiagnosis of tritrichomonosis in cats using InPouch™ TF-Feline medium. PCR-based verification of culture-positive samples prior the potentially neurotoxic ronidazole treatment is recommended.

Evaluation of the therapeutic and preventive efficacy of 2.5 % moxidectin / 10 % imidacloprid (Advocate(®), Bayer animal health) in dogs naturally infected or at risk of natural infection by Dirofilaria repens.

The objective of this GCP-compliant clinical field study was to evaluate the efficacy of the combination of moxidectin (minimum dose of 2.5 mg/kg body weight) and imidacloprid (minimum dose of 10.0 mg/kg body weight) spot-on (Advocate(®)) as a preventive and therapeutic treatment of natural infection by Dirofilaria repens in dogs in the Czech Republic.There were two arms of the study, both negatively controlled. 34 animals were randomly allocated to two groups of the treatment arm; 90 negative animals were randomly allocated to the prevention arm groups. All enrolled dogs were observed physically and blood was sampled monthly for Dirofilaria repens microfilaria counts for 18 months by modified Knott test and PCR. 34 dogs were positive for microfilaria and enrolled in the treatment arm of this study (treated: 18, untreated: 16). The reduction of the log-transformed microfilaria counts was significantly higher in the treatment group on day 28 (p = 0.007), 56, 84 and 112 (p < 0.001). All animals treated were negative after a single treatment. In the untreated control group 93.75 % remained positive (p < 0.001). 87 dogs were negative for microfilaria prior to allocation to the "preventive" arm (treated: 49; untreated: 38; 3 excluded). One dog in the untreated control group became positive for Dirofliaria repens microfilaria, while none of the treated dogs became positive. Advocate(®) was effective in the treatment of dogs infected with microfilaria of Dirofilaria repens. Due to the low rate of natural infections the preventive efficacy could not be proven, but no dog treated became positive.

Effect of therapy by using advocate spot-on combination (imidacloprid 10% and moxidectin 2.5%) on subcutaneous dirofilariosis in dogs.

Dirofilaria (Nochtiella) repens is a filarioid parasite that causes subcutaneous dirofilariosis in dogs. Adults, while localized in subcutaneous tissues, lay embryos (microfilariae (mf)) into the blood stream of dogs, which constitute a reservoir for infection of other definitive or accidental hosts as humans. This study was carried out to assess the efficacy of spot-on combination of imidacloprid and moxidectin on microfilariaemia in naturally infected dogs. A group of 11 dogs was monthly examined for the presence of microfilariae in peripheral blood by modified Knott's test method. Treatment was administered monthly for 4 months. All dogs (i.e., 100%) became negative for microfilariaemia throughout the study. These results confirm the effect of the combination of imidacloprid and moxidectin on D. (Nochtiella) repens.

Association of peripheral and central arterial wave reflections with the CYP11B2 -344C allele and sodium excretion.

OBJECTIVE: Angiotensin II and aldosterone, generated by the angiotensin-converting enzyme (ACE) and aldosterone synthase (CYP11B2), respectively, not only regulate sodium and water homeostasis, but also influence vascular remodeling in response to high blood pressure. In the European Project on Genes in Hypertension (EPOGH), we therefore investigated whether the ACE I/D and CYP11B2 C-344T polymorphisms influence early arterial wave reflections, a measure of vascular stiffness. METHODS: We measured the peripheral and central augmentation index of systolic blood pressure by applanation tonometry at the level of the radial artery in 622 subjects (160 families and 64 unrelated individuals) randomly recruited from three European populations, whose average urinary sodium excretion ranged from 196 to 245 mmol/day. In multivariate analyses, with sodium excretion analyzed as a continuous variable, we explored the phenotype-genotype associations by means of generalized estimating equations and the quantitative transmission disequilibrium test. RESULTS: The peripheral and central augmentation indexes were significantly higher in CYP11B2 -344C allele carriers than in -344T homozygotes. In offspring, early wave reflections increased with the transmission of the -344C allele. This effect of the CYP11B2 polymorphism occurred in subjects with a higher than median urinary sodium excretion (210 mmol/day). The ACE I/D polymorphism did not influence augmentation of systolic blood pressure. CONCLUSIONS: The CYP11B2 C-344T polymorphism affects arterial stiffness. However, sodium intake seems to modulate this genetic effect.

Treatment of hyperhomocysteinemia with folic acid: effects on homocysteine levels, coagulation status, and oxidative stress markers.

The aim of the study was to determine whether folic acid treatment in subjects with homocysteinemia would change their coagulation and oxidative status. Thirty-three patients with peripheral vascular disease and 26 elderly subjects with no symptoms of atherosclerosis, all of whom had total homocysteine >20 microM, were treated with folic acid (5 or 10 mg) for 3 months. In the 33 patients with peripheral vascular disease, homocysteine levels decreased from a median of 26.7 microM at baseline to 20.0 microM (p < 0.0001), whereas in the 26 asymptomatic elderly subjects, homocysteine level decreased from 24.4 microM to 18.6 microM (p < 0.0001). Plasma fibrinogen decreased whereas plasminogen and anti-thrombin increased; the differences between pre- and posttreatment values were significant in both patients and healthy subjects. Oxidative status markers showed a shift toward lower oxidative stress. This effect was observed in both study groups. An association of the therapeutic effect with the genetic polymorphism of 5,10-methylenetetrahydrofolate reductase was not detected. Folic acid supplementation to hyperhomocysteinemic subjects resulted in a decrease in total blood homocysteine concentrations; moreover, there was a tendency to reverse the coagulation status and oxidative stress.