The ability of rumen ciliates to digest chitin is clearly recognized. We investigated the chitinolytic system of the rumen ciliate Eudiplodinium maggii. The ciliates were grown in a selectively faunated sheep. They were isolated from the rumen and purified by sedimentation. A crude enzyme preparation was prepared following incubation of ciliates with antibiotics. This was done in order to reduce their contamination with intracellular bacteria. The activity of particular enzymes was examined by quantification of the products released from specific substrates. It was stated that the optimum conditions for the detected activities varied between 4.5 and 5.5 pH, and 45 and 55 °C. β-N-Acetylglucosaminidase was found as an enzyme of the highest activity (4.2 μmol/l released product per mg protein per h). The activities of endochitinase and exochitinase were almost two times lower than that of β-N-acetylglucosaminidase. Zymographic studies revealed the presence of two endochitinases, two exochitinases and two β-N-acetylglucosaminidases in the examined preparation.

• MeSH
• bachor metabolismus   mikrobiologie   MeSH
• chitin metabolismus   MeSH
• Ciliophora chemie   enzymologie   izolace a purifikace   metabolismus   MeSH
• koncentrace vodíkových iontů MeSH
• ovce MeSH
• protozoální proteiny chemie   metabolismus   MeSH
• stabilita enzymů MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Entodiniomorphid ciliates occur in the hindgut of both captive and wild African great apes. These ciliates do not form cysts, and therefore they are more susceptible for degradation. This present study focused on the survival, quantification, and decomposition processes of Troglodytella abrassarti trophozoites in the feces of captive chimpanzees. Fecal samples were examined using wet mounts and the merthiolate-iodine-formaldehyde concentration method, and the number of ciliates was expressed as ciliates per gram, which did not differ when examined from three different samples of the same feces. Trophozoites of T. abrassarti survived 5-15 hr after defecation at 25 degrees C under aerobic conditions. Decomposition of trophozoites began immediately after defecation; however, most of the trophozoites had a compact shape and visible cilia. Trophozoites, although without cilia, can be detected in the feces 55-65 hr after defecation, although most of the trophozoites were fragmented. The total number of ciliates in the sample started to decrease 35-55 hr after defecation. The absence of entodiniomorphid ciliates in fecal samples could not be caused by delayed feces fixation; instead, the absence was due to low sensitivity of coproscopic techniques. However, because of quick morphologic changes of trophozoites, accurate identification of ciliates in older samples may be difficult or even impossible.

• MeSH
• časové faktory MeSH
• Ciliophora izolace a purifikace   fyziologie   MeSH
• feces parazitologie   MeSH
• Pan troglodytes MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

We carried out a calibration of FLOTAC for ciliates Troglodytella abrassarti and Neobalantidium coli based on the selection of a most appropriate flotation solutions, and we also tested its accuracy (i.e., number of detected stages out of known added number of stages to fecal samples) and sensitivity for trophozoites of both ciliates in chimpanzee feces and N. coli cysts in pig feces, compared the detection threshold of FLOTAC with MIF-based sedimentation, and, subsequently, tested the losses of ciliate stages during sample preparation. Nine flotation solutions were evaluated, and ZnSO4 solution (specific gravity [s.g.] 1.2) showed to be the most suitable for trophozoite detection, while Sheather's solution (s.g. 1.33) was selected as most suitable for cysts. The FLOTAC sensitivity in detection of both stages varied: for trophozoites, we found all samples were positive when the intensity of infection 10 trophozoites per gram and higher, whereas for cysts the sensitivity was lower. The accuracy of FLOTAC negatively correlated with infection intensity, and the merthiolate-iodine-formaldehyde sedimentation-based quantification had a lower detection threshold. We demonstrated additional losses of stages of T. abrassarti and N. coli due to their retention in the sediment, which is probably a major reason for discrepancies in the numbers of countable ciliates between both methods. In conclusion, the FLOTAC should not be considered as a gold standard for quantification of intestinal ciliates in primates; instead, we recommend the modified MIF method.

• MeSH
• Ciliophora izolace a purifikace   MeSH
• feces parazitologie   MeSH
• infekce prvoky kmene Ciliophora diagnóza   parazitologie   veterinární   MeSH
• kalibrace MeSH
• limita detekce MeSH
• nemoci lidoopů diagnóza   parazitologie   MeSH
• nemoci prasat diagnóza   parazitologie   MeSH
• Pan troglodytes parazitologie   MeSH
• prasata MeSH
• roztoky chemie   klasifikace   MeSH
• senzitivita a specificita MeSH
• specifická hustota MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• validační studie MeSH

The ability of the rumen ciliates to utilize β-glucans other than cellulose and xylan is currently being recognized. The objective of the present study was to characterize the ability of the ciliate Diploplastron affine to digest some pachyman, laminarin, pustulan, curdlan and lichean. The protozoa were isolated from the rumen of sheep and either grown in vitro or inoculated into the rumen of ciliate-free sheep and maintained in natural conditions. In vitro culture studies showed that the enrichment of culture medium with the examined saccharides results in an increase in the number of ciliates in comparison to the control cultures. The increase was over 36 and 15 % when the growth medium was supplemented with pachyman (1,3-β-glucan) and pustulan (1,6-β-glucan), respectively. A positive correlation was also found between the population density of ciliates and the dose of saccharide supplemented to the growth medium. Enzyme studies were performed using the crude enzyme preparation obtained from ciliates treated with antibiotics. The ability of ciliates to digest the examined β-glucans was tested by the quantification of reducing sugars released from the mentioned substrates during the incubation with crude enzyme preparation. The results showed that D. affine ciliates were able to digest both of them. The mean degradation rate varied between 6.7 and 28.2 μmol/L glucose per mg protein per h for pustulan and lichean, respectively, whereas the digestion velocity was the highest at 5.0-5.5 pH and 45-50°C.

• MeSH
• bachor metabolismus   parazitologie   MeSH
• beta-glukany metabolismus   MeSH
• Ciliophora enzymologie   metabolismus   MeSH
• ovce MeSH
• protozoální proteiny metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH