Two new species of coccidia (Apicomplexa: Eimeriidae) from Madagascar Gekkonids

Two new species of coccidia (Apicomplexa: Eimeriidae) are described from the Madagascar giant day gecko, Phelsuma madagascariensis grandis, and the Golddust day gecko, P. laticauda. Both species of coccidia were found to infect the anterior one-half of the small intestine. Oocysts of Eimeria brygooi n. sp. are spherical or subspherical, 23.0 X 21.3 (18.8-25.2 X 16.4-23.2)micron; shape index (L/W) 1.1 (1.0-1.2). A micropyle, oocyst residuum, and polar granule are absent. Sporocysts are ovoid, 9.2 X 7.9 (8.0-10.0 X 7.2-8.8) micron; shape index 1.2 (1.0-1.3), with a Goussia-type suture; Stieda and substieda bodies are absent. A sporocyst residuum is present, 4.2 X 3.0 (3.2-6.4 X 2.4-4.0) micron. Sporozoites are elongate, with anterior and posterior refractile bodies. This coccidian was found to infect five of six (83%) P. m. grandis and one of five (20%) P. laticauda examined. Oocysts of Isospora gekkonis n. sp. are spherical or subspherical, 24.2 X 22.0 (21.6-26.4 X 20.0-23.6) micron; shape index 1.1 (1.0-1.2). A micropyle and oocyst residuum are absent; polar granule present. Sporocysts are ovoid, 12.2 X 9.4 (11.2-12.8 X 8.4-10.0) micron, with Stieda and substieda bodies; shape index 1.3 (1.2-1.4). A sporocyst residuum is present, either compact, 5.1 X 4.2 (4.0-7.2 X 3.2-5.6) micron or diffuse. Sporozoites are elongate, with anterior and posterior refractile bodies. Isospora gekkonis was found in two of six (33%) P. m. grandis and one of five (20%) P. laticauda. In addition, oocysts of Cryptosporidium sp. were found in the cloacas of two of six (33%) necropsied P. m. grandis.     

The selection of microbial communities by constant or fluctuating temperatures

Abstract The diversity of bacterial communities isolated from Antarctic lake sediment in chemostats under constant low temperature (8°C) or diurnally fluctuating temperature (1°C to 16°C) was examined. The median optimum temperature for growth of the freshwater bacteria isolated from the fluctuation chemostat was significantly lower ( P < 1%) than that for those from the constant temperature chemostat. The diversity of the enriched bacterial community isolated in the chemostat culture subjected to short-term temperature fluctuations was greater than that enriched under constant temperature. At least 4 different groups of bacteria, that occupied separate 'temperature niches', were isolated from the fluctuating chemostat compared to only one group isolated from the stable chemostat. Furthermore, a pseudomonad from the fluctuating chemostat was shown to out-compete another pseudomonad from the stable chemostat when both were subjected to the fluctuating temperature regime. However, the pseudomonad of constant (8°C) temperature origin out-competed that isolated under fluctuating conditions when subjected to a stable temperature regime.     

A comparative study of the development of Eimeria nieschulzi in vitro under aerobic and reducing conditions

Sporozoites of the rat coccidian, Eimeria nieschulzi Dieben, 1924 (Apicomplexa: Eimeriidae), were inoculated onto monolayers of normal rat kidney (NRK) fibroblasts and cultured either under aerobic (5% CO2/95% air) or reducing (desiccator jars modified into candle jars) conditions in RPMI-1640 supplemented with 5% fetal bovine serum, sodium bicarbonate, and antibiotics. Under aerobic conditions, first-generation meronts were observed at 2 days postinoculation (DPI) and, except for individual third-generation meronts that were seen at 5 and 6 DPI, no further development was noted. Under reducing conditions, however, first-generation meronts observed at 2-5 DPI underwent additional development to form second-generation meronts (3-5 DPI), third-generation meronts (3-7 DPI), and a small number of fourth-generation meronts (5-8 DPI). Both second- and third-generation meronts were abnormal, exhibiting gigantism although the merozoites produced appeared normal. The gradual degeneration of cell monolayers under reducing conditions prevented further observations beyond 8 DPI. These results suggest that atmospheric conditions play an important role in the development of E. nieschulzi and maintenance of reducing conditions may be one key to achieving enhanced development of some species of coccidia in vitro.     

Sporocysts isolated from the southern copperhead (Agkistrodon contortrix contortrix) produce Sarcocystis montanaensis-like sarcocysts in prairie voles (Microtus ochrogaster)

Sporulated oocysts and free sporocysts of a Sarcocystis sp. were isolated from the feces of a southern copperhead (Agkistrodon contortrix contortrix) collected in Arkansas (USA). Twenty sporocysts measured 11.2 by 8.5 microns, lacked a Stieda body, and had four sporozoites and a granular sporocyst residuum. Sarcocysts similar to those of Sarcocystis montanaensis were present in the tongues of prairie voles (Microtus ochrogaster) inoculated orally with 800 sporocysts 128 days previously. Sarcocysts were thin-walled, divided into compartments by septa, and had electron dense projections (0.14 microns) on the primary cyst wall. Infection was not pathogenic for prairie voles under the conditions of this study. No infections were observed in ICR strain laboratory mice (Mus musculus) or white-footed mice (Peromyscus leucopus) following oral inoculation of 800 sporocysts.     

Description of the oöcysts of Eimeria arnyi n. sp. (Apicomplexa: Eimeriidae) from the eastern ringneck snake Diadophis punctatus arnyi (Serpentes: Colubridae)

Coccidian oöcysts recovered from the faeces of eastern ringneck snakes, Diadophis punctatus arnyi, from Kansas, USA were found to represent a previously unreported eimerian. Oöcysts of Eimeria arnyi n. sp. are subspherical, 16.9×15.1 (15–18.5×13.5–16) m, with a thin, single-layered wall and a shape-index (length/width) of 1.1 (1.1–1.3). A micropyle and öocyst residuum are absent but a large polar granule is present. The sporocysts are elongate, 13.2×6.9 (12–14.5×6.5–7) m, with Stieda and substieda bodies and a shape-index of 1.9 (1.7–2.3). Each sporozoite contains one to two anterior and a single posterior refractile bodies. Sporulation was exogenous and complete within four days at 23°C.     

Identification and nucleotide sequence of the thymidine kinase gene of Shope fibroma virus.

The thymidine kinase (TK) gene of Shope fibroma virus (SFV), a tumorigenic leporipoxvirus, was localized within the viral genome with degenerate oligonucleotide probes. These probes were constructed to two regions of high sequence conservation between the vaccinia virus TK gene and those of several known eucaryotic cellular TK genes, including human, mouse, hamster, and chicken TK genes. The oligonucleotide probes initially localized the SFV TK gene 50 kilobases (kb) from the right terminus of the 160-kb SFV genome within the 9.5-kb BamHI-HindIII fragment E. Fine-mapping analysis indicated that the TK gene was within a 1.2-kb AvaI-HaeIII fragment, and DNA sequencing of this region revealed an open reading frame capable of encoding a polypeptide of 176 amino acids possessing considerable homology to the TK genes of the vaccinia, variola, and monkeypox orthopoxviruses and also to a variety of cellular TK genes. Homology matrix analysis and homology scores suggest that the SFV TK gene has diverged significantly from its counterpart members in the orthopoxvirus genus. Nevertheless, the presence of conserved upstream open reading frames on the 5' side of all of the poxvirus TK genes indicates a similarity of functional organization between the orthopoxviruses and leporipoxviruses. These data suggest a common ancestral origin for at least some of the unique internal regions of the leporipoxviruses and orthopoxviruses as exemplified by SFV and vaccinia virus, respectively.     

Effect of estradiol on splenic repopulation by endogenous and exogenous hemopoietic cells in irradiated mice

Estradiol treatment of irradiated mice during repopulation of their spleens by endogenous hemopoietic cells reduced the number of myelocytic colonies and increased the numbers of erythropoietic and undifferentiated colonies. The inhibitory effects of the hormone on myelopoiesis were not dependent on stimulation of erythropoiesis, since they occurred in the absence of erythropoiesis in mice made polycythemic by hypertransfusion. Treatment of bone marrow donors with estradiol reduced the ability of their marrow cells to form spleen colonies, particularly reducing the proportion of myelopoietic colonies relative to the total number of colonies of all types. Conversely erythropoietic colonies, though reduced in absolute number, were increased in relative number. Such treatment also decreased the volume and cell content of the marrow cavity through stimulation of endosteal bone formation. Estradiol treatment of lethally irradiated recipient mice did not detectably alter the total numbers or types of hemopoietic spleen colonies formed in these animals from transplanted marrow cells; however, without estradiol treatment, myelopoietic colonies were so few and erythropoietic colonies so numerous that the effects of the hormones may have been undetectable by the methods employed. The sex of the donor or recipient mouse did not affect the numbers or types of colonies formed, suggesting that endogenous levels of estradiol were too low to exert effects dectectable by the methods used. However, since our mice were only 8 weeks old, the data do not exclude the possibility that older female mice, with higher levels of estradiol, would have differed from males in relative numbers of myelopoietic as compared with erythropoietic colonies.     

Production and characterization of a compound inhibitory to Vibrio parahaemolyticus from Proteus vulgaris

The production and characteristics of a compound in Proteus vulgaris G cultures which was capable of inhibiting Vibrio parahaemolyticus and other food-borne pathogens was investigated. Production was influenced by medium composition, pH and temperature but not by the extent of aeration. The compound was most inhibitory at the optimum temperature for growth of V. parahaemolyticus. The inhibitor was most stable at pH 7·0 and inhibition occurred even after heating at 70°C for 30 min and after autoclaving. Ultrafiltration showed that the inhibitor had a molecular weight less than 1000. Thin layer chromatography of filtrates and subsequent peptidase digestion indicated that it was at least in part a peptide. The inhibitor purified by Sephadex G-15 gel filtration had a calculated molecular weight of 731 and contained only six amino acids.