| Abstract: | ABSTRACT: The purpose of this study was to: (1) determine if Histomonas meleagridis could be cultured free of bacteria, (2) produce specific antiserum and conjugate it with fluorescent dyes for use in locating and identigying H. meleagridis in tissues of various hosts. The H. meleagridis culture used for infecting turkey poults was obtained from Dr. W. M. Reid of the Poultry Science Department, University of Georgia. When the turkey poults developed signs of the disease, livers with typical histomoniasis lesions were removed aseptically, placed in a sterile Waring blender and minced in sterile physiological saline. Penicillin and dihydro-streptomycin were added to the liver suspension to eliminate contaminating bacteria. The liver suspension was injected into 10-day embryonating turkey eggs by the allatonic and yolk sac routes at the rate of 0.2 ml per egg. Eight serial passages were made in embryonating turkey eggs. Allatonic fluid collected from the eighth passage was infective for turkey poults when injected per rectum. Infected poults had gross and microscopic lesions characteristic of histomoniasis. H. meleagridis was cultured in a hamster kidney cell line (ccL 15) in the absence of bacteria using histomonad liver suspension obtained in the manner previously described. Serial passages were made free of bacteria and without loss of infectivity for turkey poults when histomonad infected hamster kidney cell culture suspension was injected per rectum. Infected turkey poults had lesions characteristic of histomoniasis. The following conclusions seem warranted from data collected during the course of this investigation: 1. H. meleagridis can be cultured in 10-day old embryonating turkey eggs and in hamster kidney cell cultures. Serial passages can be made in each system in the absence of bacteria and without loss of infectivity for turkey poults. 2. Histomonads can be visually demonstrated in the hamster kidney cell line on Leighton tube coverslips by staining them with acridine orange and observing them with a fluorescent microscope. 3. The successful cultivation of H. meleagridis free of bacteria in embryonating turkey eggs and in a hamster kidney cell line suggests the possibility of preparing an effective vaccine against histomoniasis. 4. Rabbits can be successfully used to produce antisera specific for H. meleagridis, and the labeled immune sera can be successfully used to demonstrate the presence of histomonads in the hamster kidney cell culture system by means of the fluorescent antibody technique. 5. The anti-histomonad rabbit sera can also be used to demonstrate the presence of H. meleagridis in the organs of histomonad infected turkey poults by means of the fluorescent antibody technique. 6. The specificity of labeled anti-histomonad sera in locating and identifying H. meleagridis in tissues of infected hosts by specific fluorescence under a fluorescent microscope might serve as a diagnostic and research tool in the study of the biology of H. meleagridis in various transport hosts. |
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