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Biomedical subjects

K Rohn

Publications and source records attributed to K Rohn.

13 recordsLinked to original sources

Cases of reduced cyathostomin egg-reappearance period and failure of Parascaris equorum egg count reduction following ivermectin treatment as well as survey on pyrantel efficacy on German horse farms.

In 2003 and 2004, on a total of 63 different German horse farms, a survey using the faecal egg count reduction (FECR) test was performed to investigate the efficacy of ivermectin (IVM, Ivomec) and pyrantel (PYR, Banminth) treatment against gastro-intestinal nematodes in a total of 767 horses. IVM treatment resulted in 100% reduction of the cyathostomin egg production 14 and 21 days post-treatment (d.p.t.) on 37 farms. On the remaining five farms, the mean faecal egg count reduction ranged between 97.7 and 99.9%. The mean cyathostomin FECR following PYR treatment ranged between 92.2 and 100% on the 25 farms tested. Therefore, based on the 90% FECR threshold suggested for detection of anthelmintic resistance in horses, neither IVM nor PYR anthelmintic resistance was detected. However, if the thresholds recommended for the detection of resistance in small ruminants were applied, on one and four farms signs of reduced IVM and PYR efficacy, respectively, were observed. In 2005, to further investigate these findings, the cyathostomin egg-reappearance period (ERP) following IVM treatment was examined on six selected farms, two of which were found to show less than 99.8% FECR in the previous survey. On these two latter farms, the ERP was less than 5 weeks, while on the other four it was at least 8 weeks. Earlier investigations described IVM cyathostomin ERP of at least 9 weeks. The efficacy of IVM to reduce Parascaris equorum egg excretion was also studied. On one farm in 2 consecutive years, IVM treatment did not lead to a significant reduction in P. equorum faecal egg counts in one and five young horses, respectively.

Animals↗

Frequency-specific electric response audiometry (ERA) and its clinical application in the diagnosis of hearing defects in the dog.

Reference values were established for frequency-specific electric response audiometry (ERA) in dogs on the basis of the results of ERA examinations of 200 animals with normal hearing. Air-conducting acoustic tubes with foam stoppers were used in the determination of the following: the latencies of waves I, III and V; interpeak latencies (IPL) I-III, III-V and I-V; amplitudes I and V; and the amplitude difference I-V. A frequency-specific stimulus (tone pip) was used for frequency-specific examination (1 to 4 kHz) over the entire frequency range indicated. These reference values were then used for the clinical examination of 50 dogs with hearing defects. A frequency-specific ERA was conducted and the results evaluated. These findings made it possible to draw objective conclusions about the degree, type and site of the hearing defects. Frequency-specific electric response audiometry was shown to be an important diagnostic tool for the detection of partial high- and low-frequency hearing loss and for the characterisation of hearing defects of otological, otoneurological and neurological origin.

Animals↗

[Pulmonary artery wedge pressure and heart rate measurement during pharmacological stress induction for left cardial function diagnosis in horses with and without heart disease].

In 18 horses, the pulmonary artery wedge pressure and the heart rate were measured during pharmacological stress load. 12 horses were healthy (4 trained, 8 untrained) and 6 horses had a heart disease (3 trained, 3 untrained). Pharmacological stress induction was carried out with the sympathomimetic drug dobutamine at a dosage rate of 7.5 microg/kg/min over 10 minutes of infusion. At the fourth minute, the parasympatholytic drug atropine was administered (5 microg/kg bw), and the heart rate and the pulmonary artery wedge pressure were continuously measured over 26 minutes. During sole dobutamine infusion, a significant decrease in heart rate and a significant increase in pulmonary artery wedge pressure were observed. After the application of atropine in the fourth minute, a significant increase in heart rate (from 35.7 +/- 6 up to 106 +/- 38/ min) and in pulmonary artery wedge pressure (from 15.7 +/- 3 up to 24 +/- 8.6 mmHg) were visible in the group of healthy horses. The horses with heart diseases had a significantly higher increase in both parameters (heart rate and pulmonary artery wedge pressure) with a significantly positive correlation (r = 0.7). The heart rate increased in the horses with heart diseases from 35.2 +/- 2,8 beats/min up to 132 +/- 45.7 beats/min and the pulmonary artery wedge pressure increased from 17.3 +/- 3,2 mmHg up to 32.7 +/- 13 mmHg. The cardiac status (healthy or heart disease) as well as the training level of the horses (untrained or trained) had a significant influence on the heart rate and the pulmonary artery wedge pressure. The untrained horses (healthy and heart disease) showed significantly higher values over a longer period of time than did the trained horses with the same cardiac status. Additionally the influence of pharmacological stress induction on echocardiographic parameters was investigated. The left atrial size (p = 0.015) and left ventricular diameter were significanly different in the systole (p = 0.008) and in the diastole (p = 0.001) between healthy horses and horses with heart diseases. All horses showed a positive correlation between the pulmonary artery wedge pressure and the left atrial size (r = 0.8), as well as between the left ventricular systolic (r = 0.6) and the diastolic diameter (r = 0.6). The correlation between the pulmonary artery wedge pressure and the left atrial size was nearly the same in the healthy horses (r = 0.74) and in the horses with heart diseases (r = 0.76). Regarding the training level, all untrained horses had a significantly higher correlation between the pulmonary artery wedge pressure and the left atrial size (r = 0.87) in comparison to the trained horses (r = 0.74). Particularly in the untrained horses with heart diseases, this correlation was remarcable (r = 0.99).

Animals↗

[Precision-controlled echocardiographic left ventricular function parameters by repeated measurement on three consecutive days in trained and untrained warmblood horses].

Echocardiographic measurements have to be reliable and reproducible with only a low day to day variability for detecting pathological changes of left ventricular myocardial function in horses. The day dependent fluctuation margin represents an important indicator for the reproducibility of a method. To find out the day to day variability of echocardiographic parameter in healthy warmblood horses, in the present study repeated echocardiographic measurements at three consecutive days were carried out (at the same time and by the same examiner) at 11 untrained and 7 trained warmblood horses. The horses were examined from the right hemithorax, in five different B- and M-Mode views in the long and short heart axis. The echocardiographic measurements in the three-day examination interval showed with coefficient of variation between 3.4 and 25.8% a good reproducibility. In order to calculate the precision of echocardiographic parameters (reproducibility from day to day) statistical analysis was carried out by means of a linear model with random effects. Herefor the variation between repeated measurements over three days and within the horses (day to day intraindividual variation) as well as the variation between the horses (interindividual variation) were taken into account. In dependency of the echocardiographic parameters 46.2-95.7% (median 85.7%) of the variation is caused by the difference between the horses (intraclass correlation) and 4.3-53.8% (median 14.3%) of the variation is caused by the repeated measurements on different days. With a coefficient of variance from 6.6% and an intraclass correlation of 0.957 the left ventricular diastolic diameter at the level of the papillary muscles showed the lowest variation between repeated measurements on different days. The heart rate with a coefficient of variance of 10.3% and an intraclass correlation of 0.462 showed the highest variation with 53.8% between the repeated measurements on different days. The precision of single echocardiographic measurements is confirmed. All parameters were also tested for differences between trained and untrained horses. Significant differences (p<0.05) could be found in the left ventricular diameter at the level of the apex cordis and under the mitral valves as well as in the systolic left ventricular area.

Animals↗

A Hammondia-like parasite from the European fox (Vulpes vulpes) forms biologically viable tissue cysts in cell culture.

Tissue cysts of parasites of the genus Hammondia are rarely described in naturally or experimentally infected intermediate hosts. However, ultrastructural examinations on tissue cyst stages of Hammondia sp. are needed, e.g. to compare these stages with those of Neospora caninum and other related parasites. We describe a cell culture system employed to examine the in vitro development of tissue cysts of a Hammondia sp.-like parasite (isolate FOX 2000/1) which uses the European fox as a definitive host. Cells of a diploid finite cell line from embryonal bovine heart (KH-R; CCLV, RIE 090) were infected by inoculation of sporozoites und cultivated for up to 3 months. Transmission electron microscopic examination of 17 day old cell culture material revealed the presence of cyst walls. Infected cell cultures cultivated for 2 months were used to feed a fox. Six to 13 days post infection the fox shed large numbers (n=1.2 x 10(7)) of Hammondia-sp. like oocysts which could not be distinguished from those used to infect the cell culture as determined by DNA sequencing of the internal transcribed spacer 1 and the D2/D3 domain of the large subunit ribosomal DNA. To find out the proportion of parasitophorous vacuoles that had developed into tissue cysts, the expression of bradyzoite markers was examined by probing infected cell cultures with mouse polyclonal antibodies against Toxoplasma gondii bradyzoite antigen 1 (anti-BAG1) and rat monoclonal antibodies against a cyst wall protein (mAbCC2). Nineteen and 90 days post infection all parasitophorous vacuoles in the cell cultures were positive with anti-BAG1 and mAbCC2. This shows that biologically viable (i.e. infectious) tissue cysts of a fox-derived Hammondia sp. isolate (FOX 2000/1) can be efficiently produced in this cell culture system. Since in vitro cystogenesis of dog-derived Hammondia heydorni has not been observed yet, in vitro cyst formation might be one trait to separate fox-derived Hammondia sp. from H. heydorni on a species level.

Animals↗

Effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on leukocyte count and survival rate of dogs with parvoviral enteritis.

Dogs with clinical signs consistent with parvoviral enteritis and leukopenia (total leukocyte count < 5.0 x 10(9) l(-1)) were included in this randomised double-blind study (treatment group: n = 22; control group: n = 21). The dogs in the treatment group received a subcutaneous daily injection of 10 microg kg(-1) of recombinant human granulocyte colony-stimulating factor (rhG-CSF) for 5 days. Clinical and blood investigations were performed prior to the first injection, daily during the treatment period and on the day after treatment ended, and then once more, 26 days after the first injection. During the study, no significant differences were found between the two groups with respect to survival rate (treatment group: 68 per cent; control group: 71 per cent, P > 0 4, Fisher-Test) and other clinical findings. Similarly the total leukocyte count, neutrophil count and other haematologic and biochemical parameters did not differ significantly between the groups, based on differences from initial values (P > 0 05). Consequently, the use of rhG-CSF in the treatment of dogs with parvoviral enteritis cannot be recommended.

Animals↗

Evaluation and validation of virus removal by ultrafiltration during the production of diaspirin crosslinked haemoglobin (DCLHb).

Virus retention during ultrafiltration through A/G Technology filter cartridges was investigated to characterize the removal process and validate the degree of virus titre reduction during the filtration of red blood cell haemolysates performed as part of the production of diaspirin crosslinked haemoglobin (DCLHb). When viruses were suspended in phosphate buffered saline solution, retention was greater with larger sized viruses and smaller filter pore size. Virus titre was maintained at starting levels in the filter retentate circuit during the course of filtration, suggesting that the virus removal mechanism is predominantly size exclusion. Evaluation of specific processing variables indicated that the retention of phiX174 virus was increased in the presence of red blood cell haemolysate or at high membrane crossflow rates and transmembrane pressures, while the retention of EMC virus was less sensitive to variations in these parameters. Using these results to design a validation protocol, log reduction values of >7.9 were demonstrated for the retention of human immunodeficiency virus, pseudorabies virus and bovine viral diarrhoea viruses, 7.6 for hepatitis A virus, and 4.2 for porcine parvovirus. It was also shown that the retention of viruses was maintained during repetitive use of the same filter cartridge.

Animals↗

Validation of the heat treatment step used in the production of diaspirin crosslinked hemoglobin (DCLHb) for viral inactivation.

A series of experiments was performed to assess the ability of the heat treatment step used in the manufacture of diaspirin crosslinked hemoglobin (DCLHb) to inactivate viruses. In-process solutions (reaction mixtures after the crosslinking process) from six different manufacturing lots were used as test media in a 1:680 scaled down system in which the key process parameters used in the large scale production were duplicated. The inactivation of five different viruses (Bovine Viral Diarrhea Virus, Pseudorabies Virus, Human Immunodeficiency Virus 1, Porcine Parvovirus and Hepatitis A Virus) was evaluated. Each validation experiment consisted of spiking the solution at 37 degrees C with virus, heating to 74 +/- 1 degrees C over a period of 30 minutes, holding at 74 +/- 1 degrees C for 90 minutes and cooling from 74 +/- 1 degrees C to less than 10 degrees C over a period of 30 minutes. Duplicate experiments were performed with each of the viruses with the exception of Human Immunodeficiency Virus 1, for which three experiments were performed. In each experiment samples were removed before, during, and after heating for the purpose of determining virus titer and evaluating key process parameters. The results obtained from these experiments confirmed that the key process parameters in these experiments using the scaled down test system reproduced those of the large scale manufacturing process. The results of the virus assays showed at least a 7 log reduction was accomplished by the heat treatment for each of the viruses tested.

Animals↗

Identification and characterization of penton base and pVIII protein of egg drop syndrome virus.

The nucleotide sequence and location of the penton base and pVIII genes of the egg drop syndrome (EDS) virus an avian adenovirus, were determined. The penton base gene is located at 34.8-38.8 map units. The coding sequence has a length of 1359 nt and encodes a polypeptide of 452 amino acids (aa) with a molecular weight of 51 105 Da. The amino acid sequence shows a homology of 57.3 and 55.3% to the structural protein IH of human adenovirus serotype 2 (HAd2) and fowl adenovirus serotype 1 (FAV1). The penton base protein lacks the integrin binding motifs RGD (Arg-Gly-Asp) and LDV (Leu-Asp-Val). At 65.1-67.3 map units an open reading frame of 753 nucleotides was identified which encodes the structural protein pVIH. It forms a protein of 250 aa with an expected molecular weight of 28 501 Da. Possible protease cleavage sites were identified. Amino acid homologies of 30.8 and 27.7% were found to the HAd2 and FAV1 pVIII genes. Remarkably, the overall amino acid identity with ovine adenovirus pVIII is 52%. The start codons of both genes are shifted leftwards compared with the respective structural elements on the HAd2 or FAV1 genomes which indicates a different genomic organization of the EDS virus.

Amino Acid Sequence↗

Validation of the heat treatment step used in the production of diaspirin crosslinked hemoglobin (DCLHb) for viral inactivation--effect of crosslinking.

Two experiments were performed to assess viral inactivation during the crosslinking and heat treatment steps of the DCLHb manufacturing process. Stroma free hemoglobin (SFHb) collected from a large scale manufacturing lot was tested in a 1:680 scaled down system in which the key parameters used in the manufacturing process were replicated. In the first study Porcine Parvovirus (PPV), a non-enveloped virus, was used to assess inactivation, while in the second study Bovine Viral Diarrhea Virus (BVDV), an enveloped virus, was utilized. In both experiments, the SFHb solution was deoxygenated and an aliquot of virus suspension was added. To initiate the crosslinking reaction, a solution of bis (3,5-dibromosalicyl) fumarate (DBBF) in HEPES buffer was added to the test solution. In both experiments the reaction times and the degree of crosslinking were normal. After crosslinking, the reaction mixtures were heated to 74 +/- 1 degrees C over 30 minutes, held at 74 +/- 1 degrees C for 90 minutes, and cooled to less than 10 degrees C over 30 minutes. In each experiment the degree of crosslinking of final product was 100% and yield of hemoglobin recovery was normal. Samples were removed prior to crosslinking, after crosslinking and before, during and after heat treatment for determination of virus titer and evaluation of key process parameters. The results from these experiments were consistent with those obtained from the full scale manufacturing process for the deoxygenation, crosslinking and the heat treatment step during the production of DCLHb. The results of virus assays showed that crosslinking has no effect on viruses and their subsequent inactivation by heat treatment.

Animals↗

Diaspirin crosslinked hemoglobin (DCLHb): characterization of the process and the product manufactured under GMP requirements for clinical studies.

Initiation of Clinical Trials in 1992 necessitated product manufacture under FDA Good Manufacturing Practice (GMP) requirements. To this end, all product components used were characterized to meet established requirements and processes and analytical methods were validated to confirm and document their utility and reproductibility. Manufacture of multiple 50+ liter lots of 10%g DCLHb under GMP requirements resulted in a pure and homogeneous product which reproducibly met a rigorous and complete set of product specifications. Final release testing of five consecutive GMP lots showed hemoglobin concentrations of 10.2 +/- 0.2g%, pH of 7.33 +/- 0.02 (37 degrees C), methemoglobin concentrations of 3.2 +/- 0.6%, degree of crosslinking of 99.8 +/- 0.1% and a P50 of 32.4 +/- 1.0 mmHg (37 degrees C). The mean overall yield for production of these five lots was 55 +/- 2%. Products were all endotoxin free as indicated by rabbit, cytokine and LAL testing (< 0.06 EU/mL); a critical characteristic for a molecule which may bind endotoxin. All lots were sterile as indicated by compendial testing.

Animals↗

[The use of follicle-stimulating hormone (FSH-P) for the induction of superovulation in cattle].

The clinical effectiveness of FSH-P for initiating superovulation in cattle was examined in 948 animals of the breed Deutsche Schwarzbunte and Deutsche Rotbunte of RPN Verden during 1988 and 1989. In the group of the FSH-P treated animals the number of transferable embryos was 4.4 +/- 4 per animal. The 25% percentile was 1 and the 75% percentile was 7 transferable embryos per donor. In 88 animals no embryos were found. In animals with a large number of embryos there was a changed quotient between transferable embryos and non transferable embryos per animal. A seasonal dependence in the number of transferable embryos was not found. Repeated treatment with FSH-P seemed to have no influence on the number of transferable embryos per animal. In the dose range between 6.6-9.4 mg FSH-P no differences in the number of transferable embryos per animal were found.

Animals↗