Evaluation of minimally-invasive methods to observe the cycle of the bitch in heat

Titelblatt, Inhaltsverzeichnis, Lebenslauf 1\. Einleitung und Literaturübersicht 2\. Material und Methoden 3\. Ergebnisse 4\. Diskussion 5\. Zusammenfassung/Summary 6\. LiteraturverzeichnisDas Ziel der vorliegenden Arbeit bestand darin die Eignung von gering invasiven Methoden (Messung der Körperinnentemperatur, Gewinnung von Vaginalsekret zur Bestimmung von pH-Wert und Progesterongehalt) zur Nutzung bei der Deckterminbestimmung bei der Hündin zu prüfen, bzw. deren Zusammenhang zum Zyklus der Tiere festzustellen. Außerdem wurden 3 verschiedene Testverfahren zur Blutprogesteronbestimmung verglichen. 15 läufige Hündinnen wurden im Zeitraum von September 1998 bis Mai 2000 zur Untersuchung vorgestellt. Zur Bestimmung des Östrus, des vermuteten Ovulationsbeginns und des optimalen Decktermins wurden die etablierten Methoden Prüfung des Duldungsreflexes, Beurteilung des äußeren Genitale, Vaginoskopie, Vaginalzytologie und Blutprogesteronbestimmung genutzt. Die zu prüfenden gering invasiven Methoden wurden zusätzlich angewandt. Es ergaben sich folgende Ergebnisse: Die unter 3.4.8 dargestellte Methode eignet sich zur Gewinnung von ausreichend Vaginalsekret um darin den Progesterongehalt zu bestimmen. Es zeigt sich sowohl ein Zusammenhang der ermittelten pH-Werte im Vaginalsekret, der Körperinnentemperatur, als auch des Progesterongehaltes im Vaginalsekret zum Zyklusgeschehen der Tiere. Bei allen untersuchten Hündinnen sinkt der pH-Wert im Vaginalsekret während der Läufigkeit und steigt im Folgenden wieder an. Bei 14 von 15 untersuchten Tieren geschah dieses Absinken während des Östrus. Neun Tiere zeigen einen pH-Wert-Abfall von 8,5 auf 7,5; vier Tiere von 8,5 auf 8; ein Tier von 8,5 auf 7 und ein Tier von 8 auf 7,5. Der mittlere pH-Wert-Abfall liegt bei 0,866 ± 0,286. Das Absinken des pH-Wertes im Vaginalsekret findet bei 14 von 15 Tieren in den Tagen 0 bis 2 p.o. statt und dauert im Mittel 2,333 ± 1,534 Tage, liegt also bei 93,3% der Tiere im Zeitraum der fertilen Periode der Eizelle und eignet sich somit zur Deckterminbestimmung. Die größte Übereinstimmung mit dem Zyklus der Hündin ergibt sich beim Vergleich des pH-Wertes im Vaginalsekret, wenn alle etabliereten Methoden gemeinsam zur Deckterminbestimmung genutzt werden. Bei 13 von 15 Tieren ist ein Anstieg des Progesterongehaltes im Vaginalsekret im Verlauf der Läufigkeit zu verzeichnen. Er ist bei 6 Tieren ab dem 8. Tag p.o. meßbar; bei 2 Tieren ab dem 4. Tag p.o.; bei 1 Tier an Tag 0; bei einem anderen Tier ab Tag 2 p.o.und bei einem weiteren ab Tag 3 p.o.; bei einem Tier an Tag 7 p.o. und bei einem ab Tag 9 p.o.. Bei 2 Tieren lag der Progesterongehalt während der gesamten Läufigkeit unter der Nachweisgrenze. Somit liegt bei 66,667% der untersuchten Hündinnen der Anstieg des Progesterongehaltes im Vaginalsekret außerhalb des Fertilitätszeitraumes der Eizelle. Der Nachweis des Progesterongehaltes im Vaginalsekret eignet sich nicht zur Deckterminbestimmung bei der Hündin, auch wenn er im Zyklusverlauf ansteigt. Bei allen untersuchten Hündinnen ist ein Absinken und ein Wiederanstieg der rektal gemessenen Körpertemperatur während der Läufigkeit festzustellen. Der mittlere Temperaturabfall liegt bei 0,596 ± 0,229 °C und schwankt bei den Einzeltieren zwischen 0,2°C und 1,02°C. Die mittlere rektale Temperatur hat an Tag 3 p.o. ihren Tiefstand erreicht. Der Tag der geringsten Körperinnentemperatur liegt bei allen Tieren zwischen Tag 0 und 4 p.o. Bei 66,667% der Tiere liegt er an Tag 2 bzw. 3 p.o., also zu Beginn der Fertilität. Die Messung der Körperinnentemperatur kann als Parameter zur Deckterminbestimmung bei der Hündin genutzt werden, da das Absinken derselben vor, bzw. während der fertilen Periode der Eizelle geschieht. Die größte Übereinstimmung mit der Temperaturkurve weist die Methode der Blutprogesteronbestimmung mit dem Target â canine ovulation timing kit auf, 73,334% der Hündinnen haben bei diesem Vergleich ihre geringste Körperinnentemperatur an Tag 2, bzw. 3 p.o. Der Hormonost â Test zeigt die Ovulation im Vergleich zum Target â canine ovulation timing kit später an, was vermutlich an den unterschiedlich definierten Messbereichen dieser beiden semiquantitativen Verfahren liegt. Der RIA von ICN eignet sich zur Überwachung der Lutealphase.The goal of the presented work was to test both the appropriateness of minimally-invasive methods (measuring inner-body temperature and obtaining vaginal secretions to determine pH-values and progesterone levels) to ascertain the optimal time for mating and to determine the connection of these measurements to the bitchýs cycle. In addition, three different test processes to determine blood progesterone levels were compared. Between September 1998 and May 2000 fifteen bitches in heat were presented for examination. The established methods of toleration reflex, assessment of the external genitalia, vaginoscopy, vaginal cytology and determination of blood progesterone levels were used to determine the estrus, the supposed onset of ovulation and the optimal time for mating. The minimally-invasive methods under investigation were also used. The results were as follows: The method described in 3.4.8 was appropriate to extract adequate amounts of vaginal secretion to measure progesterone levels therein. A connection to the cycle of the animals could be shown in the pH-values and progesterone levels of the vaginal secretions and in the inner-body temperatures. The pH-value in the vaginal secretions of all of the examined bitches decreased during the time they were in heat and then rose again. The decrease in pH-value occurred during estrus in 14 of 15 examined animals. Nine animals displayed a decrease in pH-value from 8,5 to 7,5; in four animals the decrease was from 8,5 to 8; in one animal the decrease was from 8,5 to 7 and in another from 8 to 7,5. The medium decrease in pH-value was 0.866 ± 0.286. The decrease in pH-value in the vaginal secretions occurred in days 0 to 2 p.o. in 14 of 15 examined animals and continued for a medium period of 2.333 ± 1.534 days, so that it occurred in 93.3% of the animals in the fertile period of their ovum. This measurement is thus appropriate for determining the optimal time for mating. Among all established methods to determine the optimal time for mating, the comparison of pH-values in vaginal secretions showed the greatest correspondence with the cycle of the bitches. An increase in progesterone in the vaginal secretions was determined in 13 of 15 animals during the period in which they were in heat. The increase was measurable beginning on the eighth day p.o. in six of the animals; beginning on the 4 th day p.o. in 2 animals, and beginning on days 0, 2, 3, 7 and 9 p.o. respectively in 5 other animals. Progesterone in the vaginal secretions of 2 animals was below detectable levels throughout the period in which they were in heat. Thus in 66.667% of the examined animals an increase in progesterone in their vaginal secretions occurred outside of the period of fertility of their ova. Determination of the progesterone levels in vaginal secretion is not appropriate to ascertain the optimal time for mating, even when the progesterone level increases during the cycle. Rectal measurements of the inner-body temperatures of the examined animals during the period in which they were in heat showed a decrease and then an increase in inner-body temperature. The medium decrease in inner-body temperature was 0.596 ± 0.229° C, and ranged between 0.2°C and 1.02°C among the individual animals. The medium rectal temperature was at its lowest point on day 3 p.o. All of the animals had their lowest inner-body temperatures between days 0 and 4 p.o. 66.667% of the animals had their lowest inner-body temperature on day 2 or 3 p.o., at the beginning of their fertility. Measurement of inner-body temperature can be used as a parameter in determining the optimal time for mating a bitch, because the decrease in inner-body temperature occurs either before or during the fertile period of the ovum. Of the examined testing processes for blood progesterone levels, the Target â canine ovulation timing kit showed the greatest concurrence with the temperature curve. In this comparison, 77.334% of the bitches had their lowest inner-body temperature on day 2 or day 3 p.o. The Hormonost â Test shows the ovulation later than the Target â canine ovulation timing kit, which is presumably a result of the differences in the defined areas of measurement of these two semi-quantitative processes. The RIA of ICN is appropriate for observing the luteal phase

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