Diversity, Roles, and Biotechnological Applications of Symbiotic Microorganisms in the Gut of Termite.

Termites are global pests and can cause serious damage to buildings, crops, and plantation forests. The symbiotic intestinal flora plays an important role in the digestion of cellulose and nitrogen in the life of termites. Termites and their symbiotic microbes in the gut form a synergistic system. These organism work together to digest lignocellulose to make the termites grow on nitrogen deficient food. In this paper, the diversity of symbiotic microorganisms in the gut of termites, including protozoan, spirochetes, actinomycetes, fungus and bacteria, and their role in the digestion of lignocellulose and also the biotechnological applications of these symbiotic microorganisms are discussed. The high efficiency lignocellulose degradation systems of symbiotic microbes in termite gut not only provided a new way of biological energy development, but also has immense prospect in the application of cellulase enzymes. In addition, the study on the symbiotic microorganisms in the gut of termites will also provide a new method for the biological control of termites by the endophytic bacteria in the gut of termites.

In vitro attenuated Histomonas meleagridis does not revert to virulence, following serial in vivo passages in turkeys or chickens.

The efficacy of vaccinating poultry against histomonosis was demonstrated recently. In the present study, the reversion to virulence and the residual pathogenicity of an in vitro attenuated, clonal strain of Histomonas meleagridis were tested in two consecutive experiments. The European Pharmcopoeia (Ph. Eur.) monograph for testing such features for coccidiosis live vaccines in chickens has served as a guideline. In the first experiment, attenuated histomonads were used in successive infection cycles in five groups of either five chickens or turkeys, respectively. All birds were killed at 14 days post infection (d.p.i.) to record lesion scores (LS) from livers and caeca. In the second experiment, the 5 times in vivo passaged histomonads were used to infect groups of 30 chickens and turkeys each, together with birds infected with virulent H. meleagridis. At three different time points 10 birds/group were killed and tissues of caeca, livers and lungs were used for PCR and immunohistochemistry (IHC) to confirm the presence of parasites. In the first experiment, various lesion scores were recordable in the livers and caeca of turkeys, with the highest LS 4 noticed once in the liver. In comparison, no lesions were seen in organs from chickens. In the second experiment, only mild lesions in the caeca of both turkeys and chickens were found. Liver lesions recorded as score 1 were noticed in just one individual of each species. PCR and IHC revealed that the attenuated and backpassaged histomonads were not present in liver samples but confined to the caeca, different to virulent H. meleagridis. Overall, no clinical signs or death occurred in turkeys or chickens inoculated orally and cloacally with 10(4) backpassaged histomonads in comparison with virulent parasites. Consequently, for the first time, the stable attenuation and safety of histomonads has been demonstrated, thus providing major implications for vaccine development.

Experiments to produce cysts in cultures of Histomonas meleagridis--the agent of histomonosis in poultry.

Experiments were done with cultured trophozoite stages of different clonal strains (Histomonas meleagridis/Turkey/Austria/2922-C6/04 and H. meleagridis/Chicken/Hungary/5009-C2/05) of H. meleagridis in order to induce a cyst formation as it is known in other intestinal parasites. It was shown that the best multiplication of H. meleagridis occurred at 40 degrees C in a full medium 199, when fetal calf serum and rice starch had been added. Under these conditions, numerous amoebic stages (8-12 microm in diameter) without and a few with flagellum were seen showing regular reproduction rates. When the conditions of culture were experimentally changed-and thus became worse-by decreasing the temperature, by deprivation of the medium from fetal calf serum and/or rice starch, and by changing the osmolarity, the pH, or the MgCl(2) concentration, many of the amoebic stages (containing starch granules) were destroyed, and several had obtained a spherical shape. If the culture conditions became even worse, smaller spherical stages occurred, which had only diameters of 4-7 microm and which appeared more condensed. Both spherical stages did not contain starch granules. All the previously seen stages disappeared constantly. Since a similar decrease of the optimal living conditions also occurs when intestinal or cloacal feces are deposited outside from the bird's body, the results obtained here may underline the interpretation that some of the formerly amoebic stages are able to become large spherical stages and later small spherical stages. The large spherical stage would be some type of precysts while the smaller ones would represent true cysts.

Light and transmission electron microscopic studies on the encystation of Histomonas meleagridis.

The study deals with the pleomorphic zooflagellate Histomonas meleagridis, which was cultivated under different stress conditions to induce a possible encystation. In the present paper, the morphological changes were analyzed by light and electron microscopy. The determination of the proliferation under different adverse conditions led to conclusions on the tenacity of the flagellate. H. meleagridis parasitizes in the intestinal tract of galliform birds and may cause enormous losses in poultry farming. For the development of new therapy approaches, clarification of the transmission pathways will be helpful. Different clonal cultures of H. meleagridis established by micromanipulation and exposed to media lacking different ingredients, inappropriate temperatures, and/or distinct reagents were investigated. Lowering of temperature was proven to have adverse effects on the survival of H. meleagridis. The flagellate could not survive in a frozen medium, and survival in a temperature of 4 degrees C lasted no longer than 23 h. An addition of sodium chloride induced an increased proliferation; pH values between 2 and 8 set limits for the survival of the parasite in different ways. H. meleagridis was able to survive under high acidic conditions for only 1 h. The major amount of cells, which could be discovered in the controls, measured 8-12 microm appeared amoebic (stage 1) and were filled with enclosures of rice starch. A rounding of most cells was noted 4 h at 4 degrees C after incubation in minimal essential medium in the absence of rice starch and fetal calf serum. A higher osmolarity of the medium, which was initiated by the addition of sodium chloride or magnesium chloride, did not induce an encystation process. After addition of hypochlorite base and cultivating at pH values between 7 and 8, spherical stages without a flagellum were formed (stage 2) measuring about 8-12 microm in diameter. Their interior consisted of a central and a peripheral region when studied by transmission electron microscopy. This aspect was due to the location of the glycogen granules. The central zone was described as totally filled with the carbohydrates, which made totally invisible the other organelles. The solidity of the amorphous layer below the cell membrane seemed to hinder the invasion of the glycogen granules. The amorphous layer below the cell membrane made it apparently possible that the cell might survive under adverse conditions-at least for a short time. This special structure might enable H. meleagridis to proceed a fast transmission and to infect many birds in a rather short time, which was shown in the past by several studies. Double-membraned cells, which were guessed to be cyst-like structures of the parasite, were also detected (stage 3). The size of these cells, however, was much smaller than that of the amoebic stages or the above-described spherical forms of H. meleagridis. Furthermore, the small cells were characterized by other granula structures. These findings might be interpreted that the small stages are possibly long-term (true) cysts and that the spherical stages with the amorphous layer beneath the cell membrane might be short-term cysts. Both, however, should be able to survive situations outside of a body and thus might be transmitted from feces to another animal.