ALBERT

Notes: Abstract. Encystation and excystation of Acanthamoeba castellanii were studied by transmission electron microscopy. The differentiation process was induced in asynchronous cultures grown axenically. Cytoplasmic vesicles containing a dense fibrous material very similar in appearance to the cyst wall were observed in trophozoites induced to encyst. When these trophozoites were incubated with calcofluor white m2r, fluorescence was observed in cytoplasmic vesicles, suggesting that the material contained in these vesicles corresponded to cyst wall precursors. Semithin cryosections of mature cysts with the same treatment showed fluorescence in the ectocyst and a less intense fluorescence in the endocyst, suggesting the presence of cellulose in both structures of the cyst wall. In mature cysts induced to excystation, small structures very similar to electron-dense granules (EDG) previously described in other amoebae were frequently observed. The EDGs were either sparsely distributed in the cytoplasm or associated with the cytoplasmic face of the plasma membrane. Many of them were located near the ostiole. In advanced phases of excystation, endocytic activity was suggested by the formation of endocytic structures and the presence of vacuoles with fibrous content similar to that of the cyst wall. Electron-dense granules in the process of dissolution were also observed in these vacuoles. Furthermore, the formation of a pseudopod suggests a displacement of the amoeba toward the ostiole.

Notes: Amoebae were isolated from a natural thermal water source in Michoacaan, Mexico, in September 1986. Two 500-ml samples were taken from pools with water at 45°C and 46°C and concentrated at 2,000 g for 15 min. The sediment was seeded on nonnutritive agar plates and incubated at 42°C. The isolates were axenized in bactocasitone-serum medium. The identification of the isolates was based on their morphology, total protein and isoenzyme patterns by agarose isoelectric focusing, serology, fine structure, agglutination with Concanavalin A, sensitivity to trimethoprim, capacity to kill mice, and their cytopathic effect in Vero cells. The results showed several morphophysiological, biochemical and serological differences between the isolates and the type strain Aq/9/1/ 45D of Naegleria lovaniensis. These remarkable differences provide sufficient evidence to consider one of the isolates a new subspecies, and the other one a morphological variant of N. l. lovaniensis, which can be differentiated from other Naegleriae by their morphology, biochemistry, serology and physiology. The authors propose the name tarasca for the subspecies and purepecha for the morphological variant.

Notes: Abstract Pathogenic and non-pathogenic free-living amoebae from an activated-sludge plant were investigated. The greatest species diversity occurred in the raw, settled sewage and at the beginning of the aeration tank, whilst the lowest number of species occurred in the final sedimentation tanks. Thirteen species of free-living amoebae representing eleven genera were isolated; these included three species of Acanthamoeba (A. castellanii, A. culbertsoni and A. polyphaga), all of wich were pathogenic to mice with varying degrees of virulence. The implications for the public health risk of the presence of pathogenic free-living amoebae in activated-sludge plants are discussed.

Notes: Abstract A protozoological survey was performed to analyze the organisms of the subphylum Sarcodina Schmarda, present in waste stabilization ponds located at Santo Tomás Atzingo, México, from March to December 1981. The amoebae isolated were identified and counted. Several physicochemical parameters were also determined and correlated with the biological data. Thirteen amoebic species were isolated and identified: one definite pathogen, Entamoeba histolytica Schaudinn; three species that have shown pathogenic strains for humans: Acanthamoeba polyphaga Page, Acanthamoeba castellanii Douglas, and Naegleria sp. (Alexeieff) Calkins; two opportunist species: Entamoeba coli (Grassi) Casagrandi and Barbagallo, and Iodamoeba bühschlii (Von Prowazek) Dobell; and seven free-living amoebae: Acanthamoeba astronyxis Ray and Hayes, Amoeba proteus (Pallas) Leidy, Hartmannella exundans Page, Hartmannella vermiformis Page, Pelomyxa palustris Leidy, Vahlkampfia avara Page, and Vahlkampfia russelli Singh. A clear correlation between the number of amoebae and the temperature and alkalinity of the system was found. The removal capacity of the ponds for E. histolytica varied from 30 to 100% during the survey. The amoebae showed a succession in space and time in the system studied.

Notes: Abstract A survey on Chromobacterium sp. and Flavobacterium sp. was conducted in a waste stabilization pond system at Santo Tomas Atzingo, State of Mexico. For this purpose methods for the isolation and identification of Chromobacterium sp. and Flavobacterium sp. from wastewater were developed and proved to be efficient. Such methods are the result of combining and modifying a series of previously reported techniques for this genera strains, but for environments different from wastewater.

Notes: Abstract Pathogenic and non-pathogenic free-living amoebae from the Root Zone Method (RZM) of wastewater treatment were investigated. Ten reed beds planted on four different substrate types were compared for their efficiency of removal of free-living amoebae. Reed beds planted on coarse substrates generally gave the highest removal rates (up to 100%) while those planted in soil gave the lowest (60.4%). Possible reasons for differences in removal efficiencies are discussed. Thirteen species representing nine genera of free-living amoebae were isolated; these included three species of Acanthamoeba (A. astronyxis, A. polyphaga and A.rhysodes) all of which were pathogenic for mice. The implications for RZM design of the presence of potentially pathogenic free-living amoebae in the effluent are discussed.

Notes: Abstract A survey was carried out over a one-year period to isolate amoebae suspended in the air of Mexico City and its suburbs. Sampling stations were placed at the four cardinal points of the metropolitan area. Selective media were used to culture the amoebae isolated. Specialized taxonomic keys and physical and physiological tests were used for identification, and a statistical analysis was performed to determine the correlations between physico-chemical and biological parameters. 108 strains were isolated, of which 19 were pathogenic via intracerebral inoculation and 9 via intranasal inoculation. Species of the generaAcanthamoeba, Vahlkampfia andHartmannella were most abundant.Acanthamoeba polyphaga showed the highest abundance. Several times during the period of the study SO2 O3, CO, NO and NO2 exceeded the permissible levels established by the Mexican government. The ability of amoebae to form cysts and cyst size were important factors for their presence, survival, abundance and diversity in the atmosphere. The main source of air-borne amoebae was the soil. Factors that favored the incidence and diversity of the isolates were wind speed and direction, low relative humidity, generation of frequent dust-storms, resuspension of amoebae by vehicular traffic, proximity to garbage dumps and large extensions of bare soil. Soil cover was a factor associated with a reduction in the incidence and diversity of the aerial amoebae. This study demonstrates that there are viable cysts of amoebae in the atmosphere of Mexico City, that may have potential importance in the case of certain kinds of human allergies and diseases. Further research is needed to find out the aerial presence of viable cysts of obligatory, amphizoic or opportunistic amoebic parasites, and to clarify the qualitative and quantitative effects of the local meteorological and physico-chemical environment on the free-living amoebae present in the atmosphere.

Notes: Abstract A survey was carried out over a 1-yr period to isolate protozoa suspended in the air of Mexico City and its suburbs. Sampling stations were placed at four cardinal points of the metropolitan area. Selective media were used to culture the protozoa isolated. Specialized taxonomic keys were used for identification and a statistical analysis was performed to determine the correlations between physicochemical and biological parameters. 63 strains were isolated. Species of Kinetoplastida and Chrysomonadida were most abundant, namely species of the genera Bodo, Cercobodo, Monas and Helkesimastix. Amoeboflagellates found included Mastigamoeba and Tetramitus. The only ciliate isolated was Colpoda steini. The greatest number of strains was yielded by Bodo repens and Cercobodo radiatus. Several times during the period of the study SO2, O3, CO, NO, and NO2 exceeded the permissible levels established by the Mexican government. From the results of this study it was concluded that the ability of protozoa to form cysts and cyst size were important factors for their presence, survival, abundance and diversity in the atmosphere. The main source of air-borne protozoa was the soil. Factors that favored the incidence and diversity of the isolates were wind speed and direction, high relative humidity, generation of frequent dust-storms, resuspension of protozoa by vehicular traffic, proximity to garbage dumps and large extensions of bare soil, and temperature (the latter only when it did not favor the production of high levels of O3). Factors that were hostile to the aerial protozoa were industry and their contaminants, and high concentrations of O3 and SO2, especially in winter time when thermal inversions occurred. Soil cover was also associated with a reduction in the incidence and diversity of the aerial protozoa. This study demonstrates that there are viable cysts of protozoa in the atmosphere of Mexico City, that may have potential importance in the case of certain kinds of human allergies and diseases. Further research is needed to find out the aerial presence of viable cysts of obligatory, amphizoic or opportunistic protozoan parasites, and to clarify the qualitative and quantitative effects of the local meteorological and physico-chemical environment on the free-living protozoa present in the atmosphere.