Zusammenfassung
Untersuchungen Über die Entwicklung einiger Nematoden-Arten in vitro hatten folgendes Ergebnis:
In modifiziertem „Ae“-Medium nach Leland (1963) (Medium H) entwickelten sich in sterilen, ständig bewegten Kulturen unter Luft als Gasphase bei 38°C die Trichostrongyliden-Arten Cooperia punctata und Ostertagia leptospicularis von der entscheideten 3. Larve bis zum geschlechtsreifen Tier, Ostertagia ostertagi bis zum 5. und Nematodirus sp. bis zum 4. Stadium. Die Entwicklung von Oesophagostomum sp. vom Schwein führte in Medium H (mit Schweineserum) unter einer Gasphase aus 5% CO2—5% O2—90% N2 bis zum 5. Stadium.
Der Einfluß verschiedener Faktoren auf die parasitische Entwicklung wurde an Cooperia punctata untersucht: In vergleichenden Prüfungen verschiedener Nährlösungen bot Medium H (Anfangs-pH 7,0–7,3; Gasphase Luft) die besten Bendingungen für die Entwicklung dieser Art. Dieses Medium bestand aus Kükenembryo-Homogenat-Extrakt, Serum, Casein-Hydrolysat, Leberextrakt, Vitaminen, Antibiotika und Earle-Lösung. In 16 von 20 Versuchen mit diesem Medium entwickelten sich 5. Stadien, in 7 dieser Experimente traten auch geschlechtsreife Exemplare auf, deren Entwicklung etwa doppelt so lang war wie im natürlichen Wirt. Die von den Weibchen in vitro abgelegten Eier waren nicht entwicklungsfähig. Für die Entwicklung von Coperia punctata waren pH-Werte von 6,0–7,6 optimal. Nach Begasung der Kulturen mit Kohlendioxyd-Luft-Gemischen entscheideten sich in Medium H mehr 3. Larven und es bildeten sich mehr 4. Stadien als unter Luft. Unter Sauerstoff fand in der Mehrzahl der Kulturen keine Entwicklung zum 4. Stadium statt, doch blieben die Larven am Leben. Dagegen starben unter Stickstoff die 3. Larven bald ab. Unter stark verminderten Sauerstoffspannungen (2; 5 und 10%) entwickelten sich 4. Larven. Aus röntgenbestrahlten (100; 80; 40 kr) Larven entstanden in Medium H weniger 5. Stadien als aus unbestrahlten Larven; der prozentuale Anteil der Weibchen nahm mit steigender Strahlendosis zu.
Summary
Experiments on the in vitro development of some nematode species gave the following results:
In modified “Ae” medium (Leland, 1963) (medium H) the trichostrongyle species Cooperia punctata and Ostertagia leptospicularis developed from exsheated 3rd stage larva to the mature stage, Ostertagia ostertagi to the 5th, and Nematodirus sp. to the 4th stage. Oesophagostomum sp. from pig was cultivated to the 5th stage.
The influence of different factors on the parasitic development of Cooperia punctata was studied in vitro: In comparative experiments with different media medium H (initial pH 7.0–7.3; gas phase air) gave the best results. The medium contained chick embryo homogenate extract, serum, caseine hydrolysate, liver extract, vitamins, antibiotics and Earle's solution. In 16 of 20 experiments with this medium a development to the 5th stage was achieved, in 7 of these experiments mature stages also occured. The time required for their development was about twice as long as in the natural host. The eggs laid by females in cultures did not develop. For the development of Cooperia punctata the optimal pH range was 6.0–7.6. In cultures incubated with carbon dioxide-air mixtures more 3rd starge larvae exsheated and more 4th stage larvae developed than under air. Under a gas phase of oxygen no development to the 4th stage took place in most cultures, but the larvae survived. Under nitrogen 3rd stage larvae soon died. In low oxygen tensions (2; 5 or 10%) a development up to the 4th stage was observed. From x-irradiated larvae (100; 80; 40 kr) fewer 5th stages developed than from normal larvae; the percentage of females increased with increasing irradiation.
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Eckert, J. In vitro-Entwicklung invasionsfähiger Nematoden-Larven zu parasitischen Stadien. Z. F. Parasitenkunde 29, 242–274 (1967). https://doi.org/10.1007/BF00260182
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DOI: https://doi.org/10.1007/BF00260182