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  • Lutz, Hans  (11)
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  • 1
    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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  • 2
    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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  • 3
    Language: English
    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 I2 (PGI2) is the most powerful endogenous inhibitor of platelet aggregation but unstable. Iloprost is a stable PGI2 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 K3-EDTA tube, and two 1.3 ml K3-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(3)/μl, which could be prevented by the addition of 1 μL (20 ng) Iloprost leading to an increase in PLT counts by a mean of 108 x10(3)/μl. CONCLUSION: This is the first study showing an anti-aggregatory effect of the PGI2-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 PGI2 or PGE1 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: Department of Farm Animals ; Chair in Veterinary Epidemiology ; 570 Life sciences; biology ; 630 Agriculture ; Feline EDTA blood; Iloprost; Platelets; Prostaglandin I2-analogue; Pseudothrombocytopenia; Platelet aggregates
    Source: University of Zurich
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  • 4
    Language: English
    Description: BACKGROUND 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. RESULTS 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. CONCLUSION 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: Department of Farm Animals ; Chair in Veterinary Epidemiology ; 570 Life sciences; biology ; 630 Agriculture
    Source: University of Zurich
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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: 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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  • 7
    In: Journal of Clinical Microbiology, 2004, Vol. 42(8), p.3775
    Description: Bovine anaplasmosis is a vector-borne disease that results in substantial economic losses in other parts of the world but so far not in northern Europe. In August 2002, a fatal disease outbreak was reported in a large dairy herd in the Swiss canton of Grisons. Diseased animals experienced fever, anorexia, agalactia, and depression. Anemia, ectoparasite infestation, and, occasionally, hemoglobinuria were observed. To determine the roles of vector-borne pathogens and to characterize the disease, blood samples were collected from all 286 animals: 50% of the cows were anemic. Upon microscopic examination of red blood cells, Anaplasma marginale inclusion bodies were found in 47% of the cows. The infection was confirmed serologically and by molecular methods. Interestingly, we also found evidence of infections with Anaplasma phagocytophilum, large Babesia and Theileria spp., and Mycoplasma wenyonii. The last two species had not previously been described in Switzerland. Anemia was significantly associated with the presence of the infectious agents detected, with the exception of A. phagocytophilum. Remarkably, concurrent infections with up to five infectious vector-borne agents were detected in 90% of the ill animals tested by PCR. We concluded that A. marginale was the major cause of the hemolytic anemia, while coinfections with other agents exacerbated the disease. This was the first severe disease outbreak associated with concurrent infections with vector-borne pathogens in alpine Switzerland; it was presumably curtailed by culling of the entire herd. It remains to be seen whether similar disease outbreaks will have to be anticipated in northern Europe in the future.
    Keywords: Babesia ; Mycoplasma Wenyonii ; Theileria ; Anaplasma Marginale ; Switzerland ; Vectors ; Pathogens ; Anaplasmosis ; Infection ; Cattle ; Cattle;
    ISSN: 0095-1137
    ISSN: 00951137
    E-ISSN: 1098660X
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  • 8
    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 ; Life Sciences ; Animal Biology ; 'Candidatus Mycoplasma Turicensis' ; Haemotropic Mycoplasma ; Transmission ; Candidatus Mycoplasma Turicensis ; Real-Time Taqman Pcr ; Seroconversion ; Veterinary Medicine
    ISSN: 0928-4249
    E-ISSN: 1297-9716
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  • 9
    Language: English
    In: Veterinary Microbiology, 2005, Vol.107(1), pp.71-79
    Description: infection in Europe has been limited to the Mediterranean and eastern countries, to Austria and to very sporadic cases in Switzerland. There are no reports of its occurrence in the countries north of Switzerland. A severe outbreak of anaplasmosis in August 2002 in a cattle farm in the canton Grisons, Switzerland, north of the Alps, with more than 300 cattle that had to be culled, came unexpected and gave reason to hypothesize presence of an increased yet undetected prevalence of in Switzerland. Randomly selected bovine serum samples collected in 1998 and 2003 were tested using a competitive inhibitory ELISA (cELISA) to test the hypothesis. Our validation of the diagnostic sensitivity and specificity of this test, done in the outbreak herd, yielded 99.2 and 83.3%, respectively, probably underestimating the true specificity. The true seroprevalence of anaplasmosis in Swiss cattle determined by cELISA was likely to be zero with upper 95% confidence limits of 2.49% in the canton Grisons and 1.17% in the rest of Switzerland, respectively, in 1998. For 2003, these estimates were even lower. There was no significant difference in apparent prevalences between 1998 and 2003. In search of a possible reservoir, three chamoises out of 46 free ranging wild ruminants from the Swiss National Park, Grisons, tested positive in the cELISA. This reaction is in accordance with or a cross reacting agent such as . From our results we conclude that the hypothesis of an increased prevalence of anaplasmosis in cattle in Switzerland must be rejected.
    Keywords: Anaplasma Marginale ; Anaplasmosis ; Cattle ; Prevalence ; Switzerland ; Celisa ; Biology ; Veterinary Medicine
    ISSN: 0378-1135
    E-ISSN: 1873-2542
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  • 10
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