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  • 1
    Language: English
    In: Veterinary Microbiology, 09 November 2012, Vol.160(1-2), pp.43-52
    Description: Hemotrophic mycoplasmas (HM) are small, cell wall-less bacteria and infections are known for a wide range of animals. One possible indication of equine HM infection was given in 1978, when a ‘haemobartonellosis’ outbreak was diagnosed in Nigerian horses by microscopy. However the first molecular proof of HM in horses was not reported until 2010, when a fragment of about 900 bp of the 16S rRNA of the equine HM was obtained. This sequence was used for the development of a SYBR green I real-time PCR assay specific for equine HM. The lower detection limit of the PCR was ten 16S rDNA copy numbers per ml of blood. The newly designed assay was successfully applied for the detection and quantification of HM in horses in Germany. A high sample prevalence of 26.5% (95% CI: 18.8–35.5%) was found (31 out of 117 horses). The mean bacterial load was 1.10 × 10 16S rDNA copy number/ml blood (range: minimum 1.05 × 10 , maximum 1.27 × 10 ). Equine HM were also detected by microscopy (Giemsa and acridine orange stained blood smears), but results do not correlate very well with PCR results, as microscopy proved rather unspecific and not sensitive. In horses younger than one year, a significant correlation between PCR positive status and anemia was found. No correlation was found in PCR-positive animals older than one year. Therefore we assume that HM infection has a higher clinical relevance in young animals.
    Keywords: Hemotrophic Mycoplasma ; Horse ; Anemia ; Real-Time Pcr ; Sybr Green ; Prevalence ; Microscopic Diagnostic ; Biology ; Veterinary Medicine
    ISSN: 0378-1135
    E-ISSN: 1873-2542
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  • 2
    Language: English
    In: BMC Veterinary Research, August 6, 2015, Vol.11(1)
    Description: Background In vitro platelet aggregation in feline blood samples is a well-known phenomenon in veterinary clinical laboratories resulting in high numbers of pseudothrombocytopenia. Several attempts have been made to prevent or dissolve platelet aggregates in feline blood samples and to increase the reliability of feline platelet counts. Prostaglandin I.sub.2 (PGI.sub.2) is the most powerful endogenous inhibitor of platelet aggregation but unstable. Iloprost is a stable PGI.sub.2 analogue. The aims of the present study were (1) to evaluate the anti-aggregatory effect of Iloprost on feline platelet counts and to determine a useful concentration to inhibit platelet aggregation in EDTA samples from clinically healthy cats, (2) to investigate the effect of Iloprost on hematological blood parameters, and (3) to determine stability of Iloprost in K3-EDTA tubes for up to 16 weeks. From 20 clinically healthy cats blood was drawn from the jugular vein and immediately distributed in a 1.3 ml K.sub.3-EDTA tube, and two 1.3 ml K.sub.3-EDTA tubes containing 20 ng and 200 ng Iloprost, respectively. A complete blood cell count was performed on the Sysmex XT-2000iV and the Mythic 18 on eight consecutive time points after collection. Blood smears were evaluated for the presence of PLT aggregates. Results In the absence of Iloprost, pseudothrombocytopenia was observed in 50 % of the investigated samples that led to significantly decreased optical PLT counts by a mean of 105 x10.sup.3/[mu]l, which could be prevented by the addition of 1 [mu]L (20 ng) Iloprost leading to an increase in PLT counts by a mean of 108 x10.sup.3/[mu]l. Conclusion This is the first study showing an anti-aggregatory effect of the PGI.sub.2-analogue Iloprost in feline EDTA blood. In all clinically healthy cats investigated, pseudothrombocytopenia was prevented by adding Iloprost to EDTA tubes prior to blood collection. Furthermore, Iloprost was very useful in preventing falsely increased WBC counts in samples with platelet aggregates analyzed on impedance-based hematological instruments. Iloprost is preferable to PGI.sub.2 or PGE.sub.1 due to its stability and easy and safe handling properties. Cytological evaluations of blood smears as well as other hematological parameters were not influenced to a clinically significant degree by the presence of Iloprost. Keywords: Feline EDTA blood, Iloprost, Platelets, Prostaglandin I.sub.2-analogue, Pseudothrombocytopenia, Platelet aggregates
    Keywords: Blood Banks – Analysis ; White Blood Cell Count – Analysis
    ISSN: 1746-6148
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  • 3
    Language: English
    In: BMC veterinary research, 05 November 2015, Vol.11, pp.276
    Description: Feline platelets are prone to clumping after blood collection, rendering the determination of accurate platelet counts difficult for clinical laboratories and resulting in a high incidence of pseudothrombocytopenia in feline haematology reports. No information is available about the kinetics of platelet aggregate formation in feline ethylenediaminetetraacetic acid blood and the course of platelet counts over a clinically relevant time period. The aim of the present study was to determine platelet counts in healthy cats over a time period of 24 h after blood collection at 9 time points; to assess potential effects of platelet aggregates, anaesthesia and bleeding conditions on feline platelets and white blood cell counts; and finally, to investigate if glucose concentration is associated with the presence of aggregates. From 30 clinically healthy cats, blood samples were analysed at 9 different time points using two different haematology instruments (using fluorescence and impedance-based flow cytometry) in the counting chamber and by blood smear evaluation. Fourteen of the 30 samples were thrombocytopenic at one to 8 time points after collection as analysed on a fluorescence flow cytometry haematology analyser. At the 24-h timepoint, all thrombocytopenic samples had returned to normal platelet counts. Seventeen of the 30 samples showed platelet aggregates in the counting chamber. Significant differences in platelet counts were associated with the presence and size of aggregates and time since bleeding. No statistically significant differences in counts were found with regard to the quality of blood collection or the use of anaesthesia. Platelet aggregation and, therefore, pseudothrombocytopenia occurred in 57 % of the investigated samples at different time points. For the first time, deaggregation of feline platelet aggregates could be demonstrated as a reversible effect of platelet aggregation. For clinical laboratories or veterinarians, it may be helpful to rerun feline samples with pseudothrombocytopenia to obtain a more reliable platelet count. The quality of blood collection seems not to be causative for platelet aggregation. Blood smear evaluation is absolutely indicated in cases when haematology instruments give PLT counts below the reference interval.
    Keywords: Blood Platelets -- Physiology ; Cats -- Blood ; Platelet Aggregation -- Physiology
    E-ISSN: 1746-6148
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  • 4
    Language: English
    In: Veterinary Microbiology, 2010, Vol.144(3), pp.525-526
    Keywords: Mycoplasma Wenyonii ; Candidatus Mycoplasma Haemobos ; Eperythrozoonteganodes ; Eperythrozoon Tuomii ; Haemobartonella Bovis ; Cattle ; Biology ; Veterinary Medicine
    ISSN: 0378-1135
    E-ISSN: 1873-2542
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  • 5
    Language: English
    In: Veterinary Microbiology, 25 February 2015, Vol.175(2-4), pp.167-178
    Description: Cats persistently infected with the gammaretrovirus feline leukemia virus (FeLV) are at risk to die within months to years from FeLV-associated disease, such as immunosuppression, anemia or lymphoma/leukemia. The integrase inhibitor raltegravir has been demonstrated to reduce FeLV replication . The aim of the present study was to investigate raltegravir for its safety and efficacy to suppress FeLV replication. The safety was tested in three naïve specified pathogen-free (SPF) cats during a 15 weeks treatment period (initially 20 mg then 40 mg orally b.i.d.). No adverse effects were noted. The efficacy was tested in seven persistently FeLV-infected SPF cats attained from 18 cats experimentally exposed to FeLV-A/Glasgow-1. The seven cats were treated during nine weeks (40 mg then 80 mg b.i.d.). Raltegravir was well tolerated even at the higher dose. A significant decrease in plasma viral RNA loads (∼5×) was found; however, after treatment termination a rebound effect was observed. Only one cat developed anti-FeLV antibodies and viral RNA loads remained decreased after treatment termination. Of note, one of the untreated FeLV-A infected cats developed fatal FeLV-C associated anemia within 5 weeks of FeLV-A infection. Moreover, progressive FeLV infection was associated with significantly lower enFeLV loads prior to infection supporting that FeLV susceptibility may be related to the genetic background of the cat. Overall, our data demonstrate the ability of raltegravir to reduce viral replication also . However, no complete control of viremia was achieved. Further investigations are needed to find an optimized treatment against FeLV. (250 words)
    Keywords: Feline Leukemia Virus ; Retrovirus ; Viral Loads ; Antiretroviral Therapy ; Immune Response, Raltegravir ; Biology ; Veterinary Medicine
    ISSN: 0378-1135
    E-ISSN: 1873-2542
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  • 6
    Language: English
    In: BMC Veterinary Research, 01 September 2017, Vol.13(1), pp.1-7
    Description: Abstract Background Glucocorticoids influence the synthesis and metabolism of catecholamines (epinephrine and norepinephrine) and metanephrines (metanephrine and normetanephrine). The aim of this study was to measure urinary catecholamines and metanephrines in dogs with hypercortisolism before and during trilostane therapy. Urine samples were collected during initial work up and during therapy with trilostane in 14 dogs with hypercortisolism and in 25 healthy dogs. Epinephrine, norepinephrine, metanephrine and normetanephrine were measured using high-pressure liquid chromatography and expressed as ratios to urinary creatinine concentration. Results Untreated dogs with hypercortisolism had significantly higher epinephrine, norepinephrine, and normetanephrine:creatinine ratios compared to healthy dogs. During trilostane therapy, urinary catecholamines and their metabolites did not decrease significantly. However, dogs with low post-ACTH cortisol concentrations during trilostane therapy had less increased epinephrine, norepinephrine and normetanephrine:creatinine ratios compared to healthy dogs. There was no correlation of urinary catecholamines and their metabolites with baseline or post-ACTH cortisol or endogenous ACTH concentrations during trilostane therapy. Conclusion Influences between steroid hormones and catecholamines seem to occur, as dogs with hypercortisolism have significantly higher urinary epinephrine, norepinephrine, and normetanephrine:creatinine ratios. Once-daily trilostane therapy does not lead to a significant decrease in catecholamines and their metabolites. Trilostane-treated dogs still have increased urinary epinephrine, norepinephrine and normetanephrine:creatinine ratios during trilostane therapy.
    Keywords: ACTH ; Metanephrines ; Pheochromocytoma ; Trilostane Therapy ; Canine ; Veterinary Medicine
    E-ISSN: 1746-6148
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  • 7
    Language: English
    In: Veterinary Microbiology, 2010, Vol.145(3), pp.351-353
    Description: Haemotrophic mycoplasmas (HM) are parasites on the surface of red blood cells and known to infect a wide range of animals. However, there are no previous evidences of HM infections in horses. In this study HM were detected for the first time in the blood of two horses suffering from poor performance, apathy, weight loss, and anaemia. Using a HM specific PCR assay and subsequent sequencing the infective agents isolated from the blood of said horses were confirmed as closely related to the HM species and ‘ Mycoplasma haemobos’.
    Keywords: Haemotrophic Mycoplasma ; Horse ; Anaemia ; 16s Rdna Pcr ; Phylogenetic Classification ; Biology ; Veterinary Medicine
    ISSN: 0378-1135
    E-ISSN: 1873-2542
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  • 8
    Language: English
    In: Clinical and Vaccine Immunology, 2010, Vol. 17(12), p.1917
    Description: In felids, three hemotropic mycoplasma species (hemoplasmas) have been described: Mycoplasma haemofelis, "Candidatus Mycoplasma haemominutum," and "Candidatus Mycoplasma turicensis." In particular, M. haemofelis may cause severe, potentially life-threatening hemolytic anemia. No routine serological assays for feline hemoplasma infections are available. Thus, the goal of our project was to identify and characterize an M. haemofelis antigen (DnaK) that subsequently could be applied as a recombinant antigen in a serological assay. The gene sequence of this protein was determined using consensus primers and blood samples from two naturally M. haemofelis-infected Swiss pet cats, an experimentally M. haemofelis-infected specific-pathogen-free cat, and a naturally M. haemofelis-infected Iberian lynx (Lynx pardinus). The M. haemofelis DnaK gene sequence showed the highest identity to an analogous protein of a porcine hemoplasma (72%). M. haemofelis DnaK was expressed recombinantly in an Escherichia coli DnaK knockout strain and purified using Ni affinity, size-exclusion, and anion-exchange chromatography. It then was biochemically and functionally characterized and showed characteristics typical for DnaKs (secondary structure profile, thermal denaturation, ATPase activity, and DnaK complementation). Moreover, its immunogenicity was assessed using serum samples from experimentally hemoplasma-infected cats. In Western blotting or enzyme-linked immunosorbent assays, it was recognized by sera from cats infected with M. haemofelis, "Ca. Mycoplasma haemominutum," and "Ca. Mycoplasma turicensis," respectively, but not from uninfected cats. This is the first description of a full-length purified recombinant feline hemoplasma antigen that can readily be applied in future pathogenesis studies and may have potential for application in a diagnostic serological test.
    Keywords: Western Blotting ; Enzyme-Linked Immunosorbent Assay ; Adenosinetriphosphatase ; Hemolytic Anemia ; Secondary Structure ; Infection ; Serological Tests ; Protein Structure ; Anion-Exchange Chromatography ; Thermal Denaturation ; Complementation ; Immunogenicity ; Dnak Protein ; Conserved Sequence ; Primers ; Amino Acid Sequence ; Mycoplasma Haemominutum ; Escherichia Coli ; Mycoplasma Haemofelis ; Lynx Lynx ; Mycoplasma ; Vaccines ; Immunology;
    ISSN: 1556-6811
    ISSN: 15566811
    ISSN: 1556679X
    E-ISSN: 1556679X
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  • 9
    Language: English
    In: Veterinary Microbiology, 09 November 2012, Vol.160(1-2), pp.227-232
    Description: belongs to haemotrophic mycoplasmas (HMs) which cause infectious anaemia in a large variety of mammals. To date, no cultivation system for or other HMs has been established. We hypothesised that could grow in classical media supplemented with nutrients ( glucose, iron-binding proteins) which are naturally available from its host environment, the porcine blood. Blood from experimentally -infected pigs was used to inoculate either standard SP-4 medium supplemented with iron-binding proteins (transferrin, haemin, and haemoglobin) or glucose-enriched Hayflick medium. A quantitative -specific real-time PCR assay was applied to determine and quantify loads weekly during 12 week-incubation. The first 2 weeks after inoculation loads decreased remarkably and then persisted at a stationary level over the observation time of 12 weeks in iron-binding protein- or glucose supplemented media variants. Scanning electron microscopic analysis of liquid sub-cultures on Hayflick agar showed small, densely-packed microcolonies of irregular cells of reduced size (0.2–0.6 μm) indicating nanotransformation. The partial 16S rDNA sequence of these cultured nanocells was 99.9% identical to . cells derived from liquid cultures interact with porcine erythrocytes by fibril-like structures. We conclude, that the modified media used for cultivation are obviously unfavourable for growth but lead to culture persistence. adapt to inappropriate culture conditions by alteration into nanoforms.
    Keywords: Haemotrophic Mycoplasma ; Culture ; Quantitative Pcr ; Scanning Electron Microscopy ; Nanotransformation ; Biology ; Veterinary Medicine
    ISSN: 0378-1135
    E-ISSN: 1873-2542
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  • 10
    Language: English
    In: Veterinary Research, 2009, Vol.40(5)
    Description: The natural transmission routes of the three feline haemotropic mycoplasmas - Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum', and 'Candidatus Mycoplasma turicensis' (CMt) - are largely unknown. Since CMt has been detected in the saliva of infected cats using PCR, we hypothesised...
    Keywords: Life Sciences ; Biochemistry, Molecular Biology ; Molecular Biology ; Life Sciences ; Genetics ; Animal Genetics ; Life Sciences ; Cellular Biology ; Life Sciences ; Cellular Biology ; Cell Behavior ; Life Sciences ; Microbiology and Parasitology ; Life Sciences ; Immunology ; Life Sciences ; Neurons and Cognition ; Life Sciences ; Santé Publique et Épidémiologie
    ISSN: 0928-4249
    E-ISSN: 1297-9716
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