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Tierärztliche Fakultät - Digitale Hochschulschriften der LMU - Teil 01/07 Podcast

Tierärztliche Fakultät - Digitale Hochschulschriften der LMU - Teil 01/07

Ludwig-Maximilians-Universität München 249 Episoden Neueste vor 21 Jahren

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Einfluss Seltener Erden in Verbindung mit phytogenen Zusatzstoffen auf Leistungsparameter beim Ferkel

11.02.2005

1 minute

In China, for 40 years Rare Earth Elements (REE) are used as growth promoters in animal production. In recent studies of our group, we were able to show that there are really ergotropic effects in piglets and in pigs, also when the animals are housed and fed under our western condition. We found an increase of daily weight gain up to 12 % and feed conversion improved too. With this knowledge in mind, we became interested in the question, if phytogenic supplements in combination with REE might influence growth parameteres in piglets synergistically. Two phytogenic compounds were used alone or in combination in several feeding studies. The first feeding experiment was done with 96 piglets of the species Deutsche Landrasse X Pietrain, 12 animals per group. Over a period of ten weeks each group has been fed either a mixture of REE (as chloride, concentration of 0 respectively 300 mg per kg feed) and / or two different phytogenic supplements (concentration of 500 respectively 900 mg per kg feed). A small positive effect of 4% has been observed under the supplementation of REE. A second experiment has been done with 48 piglets (n=6 in the control, n=7 in the treatment groups). The pigs have been fed over a period of six weeks the same concentrations of REE and the two ergotrope additives as in test one. Under the influence of REE (as citrate) and / or phytogenic additives an effect on growth and feed conversion ratio become evident, however due to heavy diarrhoea, these data have to be looked at cautiously. During the third feeding experiment, lasting seven weeks, 42 piglets grouped into six groups, have been fed a mixture of REE (as citrate, concentration of 0 respectively 200mg per kg fed) and / or the two phytogenic additives (concentration of 500 respectively 900 mg per kg feed). All five groups showed an effect on the feed conversion. The intensity of the effect was not constant over the whole time of the experiment. During the second and third week, the stongest effects have been observed. Feed conversion improved by 19% in this time period. Looking at the whole seven week period there was still a 9 % better feed conversion. REE as well as the phytogenic growth promoters have been shown to have a positive impact on feed conversion, but this effect was not statistically significant. An additive effects of REE in combination with phytogenic compounds has not been observed.

Auswirkungen von "Gentling"-Programmen auf das Verhalten von Laborratten (Wistar)

11.02.2005

1 minute

The goal of this study was to investigate whether there is a “critical period” in the fourth and fifth week of life of laboratory rats, where an effect comparable to a socialisation to people can be achieved. Preliminary tests on 6 female wistar-rats at the age of six weeks have been carried out to test different methods of non-invasive stress assessment in rats, and to develop a test procedure for the main experiments. Not all investigated methods of stress assessment were applicable for the performed study. For example it could be shown that the determination of corticosterone metabolites in the faeces pointed out a distinct circadian rhythm, but no stress-related peak after performing the planned test procedure. The determination of IgA in the faeces has not been adequate to show stress-induced changes either. The developed test procedure consisted of different behaviour tests: a modified home cage emergence test, a modified open field test, and a handtest. Additionally, a neck grip and a body-surface thermometry have been carried out, and catching has been evaluated. In main experiment I, the animals have been allocated to different cages, splitting into experimental and control groups in a genetically balanced way. The rats have been housed in groups of three under standard conditions in type-IV Makrolon cages. 24 female wistar rats of age 21 days have been used for main experiment I (“early gentling”). The animals of the experimental group have been gentled for 10 minutes each day in the fourth and fifth week of life. All animals underwent tests to assess their behaviour towards people at the beginning of the sixth, eighth, tenth and fourteenth week of life, as well as at the age of six, six and a half and nine months. Following the tests at the beginning of the fourteenth week of life and at the age of six months the same test procedure has been carried out by a person unfamiliar to the rats (test with unfamiliar person). 12 female wistar rats, which were sisters of the animals in main experiment I and had been purchased together with them at the age of 21 days, have been used for main experiment II (“late gentling”). All animals were tested for the first time at the age of 6 months. In this test, as in the subsequent test with the unfamiliar person, they served as the zero check for the animals of main experiment I. Following these tests, the animals were allocated into experimental and control groups. The six animals of the experimental group were gentled for 10 minutes each day for two weeks. After the two weeks of gentling, at the age of six and a half months, and at the age of nine months, the rats of both groups have been tested to assess their behaviour towards people. For the main experiment III (early “intensified gentling”) 24 female wistar rats of the age of 21 days have been used. The animals of the experimental group were gentled in the fourth and fifth week of life for 10 minutes twice each day. Furthermore, the animals were talked to during the gentling and recieved a food reward. The tests have been performed analogously to the tests of main experiment I. All different parts of the test procedure have been analysed separately. Furthermore five principal target figures (PTF), summarising some results, have been defined. PTF 1 summarised the audible vocalisation and the freezing behaviour during the catching tests, PTF 2 the audible vocalisation and biting during the manipulations and the neck grip as well as the abortion of the neck grip, PTF 3 the behaviour during the handtest, PTF 4 the behaviour towards the stressor in the open field and PTF 5 summarised catching from the open field, without vocalisation. High values have been rated as “tameness”. The “early gentling” of main experiment I had a distinct effect on the behaviour towards people. The animals of the experimental group showed a significantly (p

Feldstudie zur kolostralen Immunglobulin-Versorgung neugeborener Kälber in Abhängigkeit von der Verweildauer beim Muttertier

11.02.2005

1 minute

Within the scope of this study the IgG-status in 87 new born calves was investigated. All the calves were born in eight different dairy farms around southern Bavaria in Germany. The procedure of the study was to leave the newborn calves with their dam and allow them to suckle. During this retention period the dam was not milked and the calf was not additionally fed by bottle. The duration of this period was set before the calf was born by assigning it to one of three groups: group I was left 12 hours with their dams, group II 24 hours and group III 48 hours. After that time, the calves were separated from their mother, placed in individual pens and fed by bottle. Blood samples were taken 36-48 hours after birth and additionally at their seventh day of life. The serum was analysed by ELISA according to its IgG-concentration. The mean serum IgG-concentration at 48 hours in group I was 12.37 mg/ml, in group II 9.87 mg/ml and in group III 16.23 mg/ml. 58.6 % of the calves in group I, 54.6 % in group II and 80 % in group III were provided sufficiently with IgG (> 8 mg/ml). 32 calves (36.8 %) did not reach this critical value, the IgG-concetration in the serum stayed under 8 mg/ml (group I: n = 12, group II: n = 15, group III: n = 5). Half of those showed partial failure of passive transfer (pFPT; serum-IgG 4-8 mg/ml) and the other half showed failure of passive transfer (FPT; serum-IgG < 4 mg/ml). During the first week of life, the IgG-concentration in the serum of all three groups declined. The mean IgG-concentration of group I went down to 11.34 mg/ml, of group II down to 9.37 mg/ml and of group III down to 14.15 mg/ml. Time of first suckling correlated significantly negative in all three groups with the serum IgG-concentration. Suckling behaviour and therefore the intake of first colostrum was influenced by several factors. An interrelation between the course of parturition and the time of first suckling could be determined. The more complicated the parturition was, the later the calf suckled initially. For both female and male calves mean time period of first suckling was 3.2 hours p.n. The number of the dam’s lactation influenced the serum IgG-concentration in the calves positively. Heifers’calves showed a mean IgG-concentration of 7.48 mg/ml in their serum and calves of cows with more than three lactations showed a significant higher mean IgG-concentration of 14.24 mg/ml. 15 calves out of 87 (17,2 %) came down with diarrhoea, having a mean IgG-concentration of 7.73 mg/ml. Four of them even died during their first week. Those four cases of death belonged to group I and II, showing a mean IgG-concentration of 2.97 mg/ml. None of the calves in group III died during the study period. The calves not suffering from diarrhoea showed a significant higher mean IgG-concentration of 13.53 mg/ml.

Durchflusszytometrische Verfahren zur Beurteilung der Spermaqualität nach einer experimentell induzierten Hyperthermie am Bullenhoden

11.02.2005

1 minute

The aim of this study was to examine if the assessments of sperm quality can be better objectified and standardized by using flow cytometric examinations. For this investigation temporary deterioration of sperm quality was induced by a local heating of testes in bulls. The scrotal surface temperature of 4 bulls was increased during the local heating of testis by enclosing the entire scrotum for 48 hours by 6.4°C to 7.1°C. Semen was collected 3 times a week from Day -7 to Day 61 and once a week from Day 68 to Day 83 (Day 0 = the day of scrotal insulation). Each ejaculate was conventionally examined (numbers of total spermatozoa, sperm concentration, morphology and proportion of vital spermatozoa after staining with bromphenol blue nigrosin). The percentage of motile spermatozoa and progressive motile spermatozoa of motile sperm were determined with a computer-assisted motility analysis. Using flow cytometry the proportions of sperm with the following parameters were examined and analyzed: intact plasma membranes after staining with SYBR14/PI, high mitochondrial membrane potentials after staining with JC1, damaged acrosome status after staining with FITC-PNA/SYTO17/PI, and sperm with defective chromatin structure or rather with high DNA fragmentation index (DFI) by SCSA. The deterioration of sperm quality after elevating testicular temperature was correspondent to a large extent to the typical changes of the conventional sperm parameters, which result is in agreement with previous similar studies. After testicular hyperthermia (Day 0) changes of sperm quality occurred in the following sequence. There was a notable increase in secondary sperm abnormalities as well as a decrease in sperm motility on Day 9 after testicular hyperthermia. The proportions of vital spermatozoa after staining with bromphenol blue nigrosin, total sperm count and sperm concentration were decreased on Day 12 for the first time. At the same time the primary sperm abnormalities began to increase. The primary abnormalities most frequently encountered were morphological head defects of spermatozoa. Concurrent with the changes specified above the following alterations of sperm parameters were observed with flow cytometry. Beginning on Day 7 after testicular hyperthermia sperm with defective acrosome status increased. From Day 9 on sperm with intact plasma membranes and sperm with high mitochondrial membrane potentials began to decrease. On Day 12 after testicular hyperthermia the proportion of sperm with defective chromatin structure (spermatozoa with high DFI) started to increase significantly. The relationships between the proportions of defective sperm chromatin structure assessed with the SCSATM test and the proportions of sperm head defects were highly significant (r = 0.81; P < 0.0001). The proportion of sperm with a high mitochondrial membrane potential correlated positively to the sperm motility (r = 0.83; P < 0.0001). Significant correlations between the viability assessed by light microscopy and the percentages of spermatozoa with intact plasma membrane obtained by flow cytometry (r = 0.77; P < 0.0001) occurred. The increase of the proportion of the sperm with defective chromatin structure (spermatozoa with high DFI) was more clearly pronounced than that of the morphological abnormal sperm heads. This result indicates that heat stress had led to chromosome defects not only in morphologically abnormal but also in normally appearing sperm cells. In addition, morphological sperm abnormalities, especially sperm head defects, may be indicative of chromosome abnormalities also in normally appearing sperm cells of an ejaculate. This study shows that flow cytometric assessments of the ejaculate is a reliable and objective method in semen investigation. It provides additional important information about the sperm quality. In further studies, it has to be clarified, whether the prospective fertilization capacity from an ejaculate and/or the fertility of a bull can be predicted with this method more reliable than with the conventional sperm evaluation.

Evaluation des vollautomatischen Hämatologiegerätes CELL-DYN 3500 im klinischen Einsatz bei Hund und Katze

11.02.2005

1 minute

In der vorliegenden Arbeit wurden die Messergebnisse des CELL-DYN 3500, eines vollautomatischen Hämatologiesystems, hinsichtlich ihrer Zuverlässigkeit bei der Analyse von Hunde- und Katzenblutproben überprüft. Hierfür wurden folgende Untersuchungen durchgeführt und mit den Resultaten der Referenzmethoden verglichen: automatisierte Zellzahlbestimmung von Leukozyten (WBC), Erythrozyten (RBC) und Thrombozyten (PLT), Hämatokritmessung (HCT), Bestimmung der Hämoglobinkonzentration (HGB), sowie automatisierte Blutzelldifferenzierung von neutrophilen Granulozyten, Lymphozyten, Monozyten, eosinophilen und basophilen Granulozyten. Vorab wurde eine Qualitätskontrolle des Gerätes und der Referenzmethoden durch serielle Präzisionsmessungen vorgenommen und eine Untersuchung der Probenstabilität bei Lagerung angeschlossen. Der CELL-DYN 3500 ist ein Multi-Parameter Durchflusszytometer, der Leukozyten (WBC) nach dem Prinzip der Laserlichtstreuung (Multi-Angle Polarized Scatter Separation; M.A.P.S.S.) sowohl zählt als auch differenziert. Erythrozyten (RBC) und Thrombozyten (PLT) werden nach dem Widerstandsmessprinzip ermittelt, nach dem zusätzlich auch die Leukozyten bestimmt werden. Die Referenzmethoden wurden nach den Empfehlungen des International Committee for Standardization in Haematology (ICSH 1984) gewählt und beinhalteten manuelle Zählungen der WBC, RBC und PLT, die Mikro-Hämatokrit-Zentrifugen Methode und die spektrophotometrische Messung der Hämoglobinkonzentration nach dem WHO Standard für Hämoglobinbestimmungen. Die automatischen Differentialblutbilder wurden mit mikroskopischen 400-Zell Differentialblutbildern verglichen. Die hohe Präzision des CELL-DYN 3500 konnte durch niedrige Variations-koeffizienten dokumentiert werden. Diese waren bei allen untersuchten Parametern durchweg kleiner als die der manuellen Referenzmethoden (Präzision in Serie). Zur Gewährleistung verlässlicher Werte der automatischen Blutanalyse sollte die Blutprobe gekühlt und innerhalb von 48 Stunden nach Blutentnahme untersucht werden (Untersuchung zur Probenlagerung). Es konnten folgende Korrelationskoeffizienten (r) durch lineare Regressionsanalyse nach Pearson ermittelt werden: 0,988 und 0,977 für WBC; 0,927 und 0,960 für RBC; 0,949 und 0,598 für PLT; 0,971 und 0,957 für HGB; 0,979 und 0,969 für HCT bei Hunden bzw. Katzen. Die Korrelationskoeffizienten für neutrophile Granulozyten waren 0,974 und 0,984, die für Lymphozyten 0,701 und 0,891 bei Hunden bzw. Katzen. Da Monozyten und insbesondere basophile Granulozyten nur in sehr geringen Konzentrationen im Blut vorliegen waren nur mäßige Korrelationen dieser Zellen zu ermitteln. Auf die statistische Auswertung der Basophilen wurde aus diesem Grund gänzlich verzichtet. Die Korrelation der eosinophilen Granulozyten war mit Korrelationskoeffizienten von 0,835 und 0,928 bei Hunden bzw. Katzen trotz niedriger absoluter Zellzahlen hoch. Dies belegte die besondere Fähigkeit des CELL DYN 3500 diese Zellpopulation richtig zu erkennen. Da der Korrelationskoeffizient (r) nur den linearen Zusammenhang zwischen zwei Methoden ausdrückt und keine Aussage über die Übereinstimmung der Messwerte trifft, wurden absolute Differenzen nach der Methode nach BLAND und ALTMAN (1986) gebildet und in einem separaten Streudiagramm graphisch dargestellt. Die Mittelwerte der absoluten Differenzen (mittlere Abweichungen) waren für sämtliche Parameter mit Ausnahmen der felinen Thrombozyten sehr gering. Es konnten so keine systematischen klinisch relevanten Abweichungen festgestellt werden, nur einzelne zufällige, nicht erklärbare. Die Ergebnisse der Thrombozytenmessungen bei der Katze sollten nicht vom Gerät übernommen werden. Die Thrombozytenmessung bei der Katze sollte wahrscheinlich grundsätzlich nicht durch Impedanzmessgeräte erfolgen. Insgesamt betrachtet, kann der CELL-DYN 3500 als ein sehr zuverlässiges und einfach zu bedienendes Gerät angesehen werden, das präzise und akkurate Messergebnisse bei physiologischen und den meisten pathologischen Blutproben von Hunden und Katzen liefert. Das Gerät kann die Bearbeitungszeit der Blutprobenanalyse signifikant verkürzen, sodass sich der Benutzer intensiver mit der Studie pathologischer Proben befassen kann. Für pathologische Blutbilder bleibt die mikroskopische Untersuchung unersetzlich. Wir betrachten jedes Blutbild, bei dem ein Parameter außerhalb des Referenzbereichs liegt, oder dessen Ergebnisse klinisch nicht plausibel sind, als mikroskopisch zu überprüfen. Knapp 40 % aller Katzen- und gut 20 % aller Hundeblutbilder werden in der Medizinischen Kleintierklinik mikroskopisch nachdifferenziert. Darüber hinaus sind sämtliche Gerätewarnungen bezüglich pathologischer Zellen, wie Blasten, unreife Granulozyten, reaktive Lymphozyten und andere, im Veterinärprogramm des CELL-DYN 3500 deaktiviert, sodass auch hier im klinischen Verdachtsfall eine mikroskopische Blutzelldifferenzierung unerlässlich ist.

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Tierärztliche Fakultät - Digitale Hochschulschriften der LMU - Teil 01/07

Episoden

Einfluss Seltener Erden in Verbindung mit phytogenen Zusatzstoffen auf Leistungsparameter beim Ferkel

Auswirkungen von "Gentling"-Programmen auf das Verhalten von Laborratten (Wistar)

Feldstudie zur kolostralen Immunglobulin-Versorgung neugeborener Kälber in Abhängigkeit von der Verweildauer beim Muttertier

Durchflusszytometrische Verfahren zur Beurteilung der Spermaqualität nach einer experimentell induzierten Hyperthermie am Bullenhoden

Evaluation des vollautomatischen Hämatologiegerätes CELL-DYN 3500 im klinischen Einsatz bei Hund und Katze

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