The Oocysts of Eimeria vermiformis sp. n. and E. papillata sp. n. (Protozoa: Eimeriidae) from the Mouse Mus musclus

SYNOPSIS. Eimeria vermiformis sp. n. and E. papillata sp. n. are described from the mouse Mus musculus. The sporulated oocysts of E. vermiformis are 18–26 by 15–21 μ (mean 23.1 by 18.4 μ); its sporocysts are 11–14 by 6–10 μ (mean 12.8 by 7.9 p). The sporulated oocysts of E. papillata are 18–26 by 16–24 μ (mean 22.4 by 19.2 μ); its sporocysts are 10–13 by 6–9 μ (mean 11.2 by 8.0 μ). A substiedal body is present in E. papillata sporocysts. Patent infections were produced in white laboratory mice with both species. Fourteen species of Eimeria have now been described from the genus Mus.     

Automated dead-end ultrafiltration of large volume water samples to enable detection of low-level targets and reduce sample variability

Aims

A Portable Multi‐use Automated Concentration System (PMACS) concentrates micro‐organisms from large volumes of water through automated dead‐end ultrafiltration and backflushing. The ability to detect microbial targets from ground, surface and cooling tower waters collected using standard methods was compared with samples from the PMACS in this study.

Methods and Results

PMACS (100 l) and standard grab samples (100–500 ml) were collected from sites in Florida and South Carolina, USA. Samples were analysed for the presence of faecal indicator bacteria (FIB; ground and surface water) or Legionella pneumophila (Lp; cooling tower water). FIB were enumerated by growth on selective media following membrane filtration or in IDEXX defined substrate media. Lp cells were detected by direct fluorescence immunoassay using FITC‐labelled monoclonal antibodies targeting serogroups 1, 2, 4 and 6. FIB were found in PMACS samples from ground and surface waters when their concentrations were below detection limits in grab samples. The concentrations of Lp in cooling tower samples collected over 5 months were more consistent in PMACS samples than grab samples.

Conclusions

These data demonstrate that PMACS concentration is advantageous for water monitoring. FIB were detected in PMACS samples when their concentrations were below the detection limits of the standard methods used. PMACS processing provided more representative samples of cooling tower waters reducing sample variability during long‐term monitoring.

Significance and Impact of the Study

This study highlights the utility of PMACS processing for enhanced monitoring of water for low‐level microbial targets and for reducing sample variability in long‐term monitoring programmes.     

Eimeria lophortygis n. sp. and E. okanaganensis n. sp. (Sporozoa: Eimeriidae) from California Quail Lophortyx californicus in British Columbia*

SYNOPSIS. Eimeria lophortygis n. sp. and E. okanaganensis n. sp. are described from California quail Lophortyx californicus in British Columbia. Oocysts of E. lophortygis are subspherical to ovoid, 19–26 by 18–20.5 μ (mean, 22.5 by 18.7 μ); sporocysts ovoid, 13–14 by 6–8 μ (mean, 13.7 by 7.5 μ). Prepatent period 72 hours. Oocysts of E. okanaganensis are ovoid to ellipsoidal, 22.5-30 by 17-21 μ (mean, 26.3 by 19.8 μ); sporocysts ellipsoidal, 13-18 by 6-8 μ (mean, 14.0 by 6.5 μ). Prepatent period 104 hours. Cross-transmission attempts with Japanese quail Coturnix coturnix and Leghorn chicks were unsuccessful.     

A comparative study of seven grabs used for sampling benthic macroinvertebrates in rivers

SUMMARY. After considering the large number of grabs described in the literature, seven grabs of weight < 25 kg were chosen for manual operation from a small boat: Van-Veen grab, weighted and unweighted Ponar grabs, Friedinger version of the Petersen grab, Dietz-La Fond mud-snapper, pole-operated Birge-Ekman grab and pole-operated Allan grab. Random samples (number of sampling units n= 10) were taken in a large tank with a known number of 2-mm cylindrical plastic pellets amongst stones of uniform size. Separate experiments were performed with four sizes of stones (model ranges: 2–4 mm, 8–16 mm, 16–32 mm, 32–64 mm). Stratified random samples (n= 10) were taken in rivers and the modal particle sizes at four sites were 0.004–0.06 mm, 0.5–2 mm, 16–64 mm and 64–128 mm. All grabs usually took a representative sample of the substratum at each site with no strong bias towards a particular particle size. The general performance of the Friedinger, Dietz-La Fond and Allan grabs was poor, except on a muddy bottom, with frequent failure to operate, small samples of substratum and a mean depth of penetration < 3 cm in all substrata except mud for the Dietz-La Fond and Allan grabs. The Van-Veen and Birge-Ekman grabs sampled to a mean depth < 3 cm in mud and fine gravel (2–4 mm), but the Birge-Ekman jammed frequently in fine gravel. Both Ponar grabs operated well and sampled to a mean depth ≥ 5 cm in mud and fine gravel, > 3 cm when small stones (8–16 mm) were present and 2 cm (weighted Ponar only) when larger stones (> 16 mm) were present in a gravel bottom. The mean depth was <0.8 cm for all grabs when larger stones (>16 mm) were predominant on the bottom. In the tank experiments with pellets, the efficiencies for the total catches of the Friedinger, Dietz-La Fond and Allan grabs were low with values <45% for fine gravel (2–4 mm), < 22% for small stones (8–16 mm) and <5% for a substratum of larger stones (>16 mm). If 50% is the minimum acceptable efficiency, then the Ponar, Van-Veen and Birge-Ekman grabs were adequate for fine gravel, only the two Ponar grabs were adequate for small stones and no grabs were adequate for sampling a substratum of larger stones (>16 mm). In field trials, the relative abundances of major taxa were similar for most grabs at each site; Friedinger and Dietz-La Fond grabs were the major exceptions. In terms of both mean number of taxa and mean number of invertebrates m^?2 the Ponar, Birge-Ekman and Allan grabs performed well on the predominantly muddy substratum at site 1, but only the weighted Ponar grab performed adequately on the predominantly gravel bottom with some large stones (>16 mm) at site 2. All grabs performed badly when larger stones (>16 mm) were predominant on the bottom (sites 3, 4). The relationship between the variances and means of the samples taken with each grab followed a power law for the catches of pellets in tank experiments, and for major taxa and total numbers at each site in field trials. Values of exponents in the power law lay within the range 1.14–2.34. The coefficient of variation was also frequently related to the sample mean and was an unreliable statistic for comparing the precision of grabs.     

Development of Haemogregarina boueti in the toad Bufo regularis*

SYNOPSIS. Haemogregarina boueti França, 1910, was found to be the commonest blood parasite in the common toad, Bufo regularis Reuss, in Egypt. The rate of infection was about 30% (of 689 toads examined). In properly fixed blood films, the parasites were almost exclusively intraerythrocytic. Most characteristic was the encapsulated “elongate” form averaging 22.3 by 6 μ with a more-or-less central nucleus and a pointed, slightly bent, posterior end. Infected red cells were conspicuously hypertrophied and their nuclei were markedly displaced and frequently broken into 2-4 parts. Young and growing blood forms as well as two types of hepatic schizonts are described for the first time. Schizonts of the first type develop in hepatic cells, are 28–30 μ in diameter and produce numerous elongate oval merozoites about 8 × 2.2 μ radially arranged around a residual body about 10 μ in diameter. Schizonts of the second type start their growth in erythrocytes but later complete their development as free bodies in the liver sinusoids. When mature, they are 32–35 μ in diameter and produce a larger number of thin merozoites about 8 × 1.5 μ, surrounding a larger residual body about 19 μ in diameter.     

Eimeria montanaensis n. sp. and E. ernesti n. sp. (Protozoa,Eimeriidae) from the Rocky Mountain Goat Oreamnos americanus*

SYNOPSIS. Oocysts of Eimeria oreamni, E. montanaensis n. sp. and E. ernsti n. sp. were present in a fecal sample collected from a male Rocky Mountain goat Oreamnos americanus in Ravalli County, Montana. Oocysts of E. oreamni were nearly identical to those originally described from this host. Oocysts of E. montanaensis were 15-23 by 13-19 μ (mean 18.9 by 15.2 μ), with sporocysts 8-12 by 4–7 μ(mean 9.9 by 5.2 μ). Oocysts of E. ernsti were 28-37 by 19-26 μ (mean 32.9 by 23.0 μ), with sporocysts 14-20 by 6-9 μ (mean 16.7 by 7.3 μ). A distinct micropyle and micropylar cap were present in E. ernsti.     

Eimeria macusaniensis n. sp. (Protozoa: Eimeriidae) of the Alpaca Lama pacos

SYNOPSIS. A new species of coccidium, Eimeria macusaniensis n. sp., is described from the alpaca Lama pacos in Macusani, Peru. The oocysts are ovoid to piriform, 81–107 by 61–80 μ (mean 93.6 by 67.4 μ) with a micropylar cap and elongate ovoid sporocysts 33–40 by 16–20 μ (mean 36.3 by 18.3 μ).     

The structure of the hemocytes of Galleria mellonella (Lepidoptera)

Four hemocyte types have been identified in the late last larval instar of Galleria mellonella. Plasmatocytoids are round to spindle shaped cells, 10–20 μ long and 5–10 μ wide. The cytoplasm contains no distinguishing inclusions. Golgi complexes, rough endoplasmic reticulum and free ribosomes are abundant. Granular hemocytes are oval shaped cells, 10–20 μ long and 5–10 μ wide. The granules, their most characteristic feature, have a diameter of 0.2 μ, a microtubular sub-structure, and are made up of acidic mucosubstances. Lipid droplets may be present in these cells at some stage of development. These cells appear to be phagocytic. Spherule cells are oval shaped, 15–20 μ long and 5–10 μ wide. The spherules, approximately 2 μ in diameter, have a highly ordered substructure and are made up of acidic mucosubstances. Oenocytoids are the largest cells, 20 by 40 μ. The cytoplasm contains mostly free ribosomes and microtubules.     

Eimeria kinsellai sp. n. (Protozoa: Eimeriidae) in a Marsh Rice Rat Oryzomys palustris from Florida*

SYNOPSIS. Eimeria kinsellai sp. n. is described from the marsh rice rat Oryzomys palustris from Florida. Its ellipsoidal oocysts were 23–31 by 15–19 μ (mean 26.7 by 16.9 μ). One to four polar granules were present but an oocyst residuum was absent. The sporocysts were 12–15 by 7–9 μ (mean 13.9 by 8.1 μ), and sporocyst residua were present.     

Isospora Species of the Cat and Attempted Transmission of I. felis Wenyon, 1923 from the Cat to the Dog*

SYNOPSIS Fecal samples from 130 domestic cats from Illinois were examined for coccidia. Three species of Isospora were found: (1) I. felis Wenyon, 1923, with oocysts 38-51 by 27-39 μ with a mean of 41.6 by 30.5 μ and sporocysts 20-26 by 17-22 μ with a mean of 22.6 by 18.4 μ; it was found in 13% of the cats; (2) I. rivolta (Grassi, 1879) Wenyon, 1923, with oocysts 21-28 by 18-23 μ with a mean of 25.0 by 21.1 μ, and sporocysts 14-16 by 10-13 μ with a mean of 15.2 by 11.6 μ; it was found in 3% of the cats; and (3) I. bigemina (Stiles, 1891) Lühe, 1906, with oocysts 12-15 by 10-13 μ with a mean of 13.2 by 11.8 μ. and sporocysts 8-10 by 6-8 μ with a mean of 8.8 by 6.5 μ it was found in 1.5% of the cats. Four coccidia-free puppies 1.5 months old were inoculated with 100,000 oocysts each of I. felis from the cat, but patent infections did not occur. Partial development of I. felis was not seen in tissue sections of the small intestine of a 5th pup killed 96 hours after inoculation with 150,000 I. felis oocysts. This coccidium is therefore presumably host-specific.     

LIFE CYCLE OF A STEPHANODISCUS SP. (BACILLARIOPHYTA)1

Studies of the life cycle of a centric diatom, tentatively identified as Stephanodiscus neoastraea Håkansson & Hickel, showed that sexual reproduction occurred every year in a freshwater lake (Lough Neagh, Northern Ireland). Male and female gametes were produced in cells below 55% of the maximum diameter during a 3–4-week period in late summer, following the return of nitrate concentrations above 10 μM NO_3-N. The frequency of sexual reproduction was linked to the cycle of diameter size reduction and regeneration. The times of largest decreases in cell diameter were during nutrient stress in summer and low light conditions in late autumn, rather than during the main spring growth period. So, environmental conditions (combined with the limited life-spans of individual cells) affected the rate of diameter reduction and, therefore, the length of the life cycle (3–4 years).     

Lankesteria barretti n. sp. (Eugregarinida,Diplocystidae), a Parasite of the Mosquito Aedes triseriatus (Say) and a Review of the Genus Lankesteria Mingazzini*

SYNOPSIS. Lankesteria barretti n. sp. (Eugregarinida, Diplocystidae) is named from the mosquito Aedes triseriatus in Texas. The young cephalins occur within the midgut epithelial cells. When they reach a length of about 150–200 μ they rupture the host cell and are released into the space between the epithelium and the peritrophic membrane, becoming gamonts. These grow to about 140–310 by 40 μ at the anterior end and 12 μ at the posterior end. When the host pupates they enter the lumen of the Malpighian tubules; pairs join in syzygy by their anterior ends and later more laterally. Each pair forms a spherical gametocyst about 60–100 μ (exceptionally 250 μ) in diameter. A large number of oocysts develop in each gametocyst. The mature oocysts are spindle-shaped, 11 by 5.4–5.7 μ, and contain 8 elongate sporozoites and a refractile residuum. The gametocyst wall breaks down, releasing oocysts in the Malpighian tubules of the host when it is adult. The oocysts pass out in the feces and presumably infect new larvae by ingestion. The cephalins and gamonts of L. barretti differ from those of L. culicis (of Aedes aegypti) in having a relatively anterior instead of a central nucleus and in lacking a noticeable mucron; its longitudinal folds are not as well-developed as in L. culicis, and its paraglycogen granules are larger. The fine structure of L. culicis and L. barretti is described in detail. Their gamonts have a polar ring but no definite conoid. The taxonomy of the genus is reviewed, but its species have been so poorly described that it is impossible to be sure whether they are all really Lankesteria. About 19 species have been described (5 from turbellaria, 8 from tunicates, perhaps 1 from Amphioxus, 1 from the chaetognath Sagitta sp., 1 from Phlebotomus and 3 from mosquitoes).     

Coccidia (Protozoa: Eimeriidae) of the Alpaca Lama pacos*

SYNOPSIS. Three new species of coccidia are described from the alpaca Lama pacos from Peru. These are the first species of coccidia to be named from this host. The oocysts of Eimeria lamae n. sp. are ellipsoidal, occasionally ovoid, 30–40 by 21–30 μ (mean 35.6 by 24.5 μ) with elongate ovoid sporocysts 13–16 by 8–10 μ (mean 15.3 by 8.5 μ). The oocysts of Eimeria alpacae n. sp. are ellipsoidal, rarely ovoid, 22–26 by 18–21 μ (mean 24.1 by 19.6 μ), with ovoid sporocysts 10–13 by 7–8 μ (mean 11.0 by 6.8 μ). The oocysts of Eimeria punoensis n. sp. are ellipsoidal, occasionally ovoid, 17–22 by 14–18 μ (mean 19.9 by 16.4 μ), with ovoid sporocysts 8–11 by 5–7 μ (mean 9.2 by 6.1 μ).     

Characterization of 2,4-Dinitrotoluene Dioxygenase from Recombinant Esherichia coli Strain PFJS39: Its Direct Interaction with Vitreoscilla Hemoglobin

Burkholderia dinitrotoluene (DNT) dioxygenase in this study (from recombinant Esherichia coli strain PFJS39) is probably a multicomponent enzyme system that oxidizes 2,4-dinitrotoluene (DNT) to 4-methyl-5-nitrocatechol (MNC). DNT dioxygenase was purified by a four-step procedure that utilized consecutive gel filtration chromatography and a nondenaturing gel system. The purified enzyme oxidized DNT only in the presence of NADH and its yield increased by lipase pretreatment of crude cytosol. An estimated molecular weight of 100,000 was obtained by gel filtration. Polyacrylamide gel electrophoresis (PAGE) in the presence of sodium dodecyl sulfate (SDS) revealed the presence of three subunits for the samples from consecutive gel filtration chromatography and nondenaturing PAGE. Their molecular weights were 52,000–71,000, 23,000–25,500, and 12,000–16,500. These results suggest that DNT dioxygenase exists as a heterotrimer. The K _M of DNT dioxygenase for O₂ is 50 μ M, consistent with inhibition results of DNT dioxygenase by Vitreoscilla hemoglobin (its K _M for O₂ is 7 μ M). The K _M for DNT is 180 μ M. The purified enzyme is relatively stable below 40°C, retains activity over a broad pH range, and is stimulated by several cofactors in addition to NADH.     

Haemogregarina faiyumensis n. sp. in the Toad Bufo regularis in Egypt*

SYNOPSIS. Haemogregarina faiyumensis n. sp., a parasite of toads, Bufo regularis, in Faiyuni Province, Egypt, U.A.R., is described. In a survey of 689 toads from various localities in Cairo, Giza and Faiyurn provinces, only 3 out of 13 toads from Kom O Shim near Faiyum were infected. This species, known only by blood forms (most probably gametocytes) of two different staining reactions, is 13-17 μ long and 4-5 μ wide, with an average of 15.5 × 4.5 μ. The nucleus is typically subcircular and 3-5 × 3–5 μ, with an average of 4.5 × 3.9 μ.     

Four New Species of Eimeria (Protozoa: Eimeriidae) from the Pika Ochotona princeps from Alberta and O. pallasi from Kazakhstan1

SYNOPSIS. Eimeria worleyi n. sp. and Eimeria barretti n. sp. from the pika Ochotona princeps from Alberta and Eimeria pallasi n. sp. and Eimeria shubini n. sp. from the pika Ochotona pallasi from Central Kazakhstan are described. Oocysts of E. worleyi were 12–16 by 10–15 μ (mean 13.5 by 12.5 μ) and spherical to subspherical. E. barretti oocysts were ellipsoidal to slightly ovoid and 27–36 by 21–27 μ (mean 32.9 by 23.8 μ). Oocysts of E. pallasi were ellipsoidal or ovoid and 19–34 by 17–26 μ (mean 26.3 by 21.3 μ). E. shubini oocysts were spherical and 22 μ in diameter. E. kriygsmanni Yakimoff and Gousseff, 1938 of Svanbaev (1958) in O. pallasi, [non] E. kriigsmanni Yakimoff and Gousseff, 1938 in Mus musculus, is a synonym of E. pallasi. E. musculi Yakimoff and Gousseff, 1938 of Svanbaev (1958) in O. pallasi, [non] E. musculi Yakimoff and Gousseff, 1938 in Mus musculus, is a synonym of E. shubini.     

UNEXPECTED RESPONSE TO VITAMIN B12 OF DOMINANT CENTRIC DIATOMS FROM THE SPRING BLOOM IN THE GULF OF MAINE (NORTHEAST ATLANTIC OCEAN)1,2

Twelve clones (seven species) representative of centric diatoms dominant in the spring phytoplankton bloom in the Gulf of Maine were isolated and rendered axenic. Genera included were Thalassiosira, Porosira and Chaetoceros. Unlike most centric diatoms studied previously, none of these has an absolute requirement for vitamin B_12. However, B₁₂ (5 ng.l^-1) stimulated growth of most clones by eliminating or reducing the lag phase and increasing the growth rate. Bloom population densities developed 4–54 days earlier with B₁₂ present. Several clones grown with B₁₂ removed more than 80% of the vitamin from the medium. When grown in vitamin-free medium the cells put 0.01–0.7 ng.l^-1 B₁₂ into the medium. We conclude that vitamin B₁₂ is of ecological significance even though the requirement for it is not absolute.     

A comparative study of three air-lift samplers used for sampling benthic macro-invertebrates in rivers

SUMMARY. From 16 air-lift samplers described in the literature, three were selected for operation from a small boat: Mackey, Pearson et al. and Verollet & Tachet samplers. Random samples (number of sampling units n= 10) were taken in a large tank with a known number of dried peas, representing invertebrates, amongst stones of uniform size. Separate experiments were performed with three sizes of stones (modal sizes 2–4, 16–20 and 32–36 mm). Stratified-random samples (usually n= 10) were taken in rivers at lour sites with modal particle sizes of < 0.1 2–4. 32–128 and 64–128 mm. The samplers were compared with a Ponar grab in the tank experiments and the field, and with a Naturalist's dredge in the field. Rates of pumping at different air-flows were established for each sampler. Water-How increased with increasing air-How and increasing submergence of the riser-pipe. The Mackey sampler achieved the highest water-flow and lifted more substratum and larger stones (64–128 mm) than the other samplers. The Verollet & Tachet sampler had the lowest pumping rate and failed to lift stones > 8 mm long, whilst the Pearson et al. sampler, with an intermediate performance, lifted mud and stones < 32 mm long. In the tank experiments, the area from which the Mackey and Pearson et al. samplers collected peas varied with air-flow, operating time and type of substratum. The Maekey sampler over-sampled and the Pearson et al. sampler usually under-sampled their respective sampling areas, except the latter sampler at high air-flows gave approximately quantitative results comparable to those of the Ponar grab for peas on the surface of stones 16–20 mm. No sampler collected much at a depth of 3 cm on larger stones (32–36 mm), and the Verollet & Tachet sampler failed completely on all substrata. In field trials, the Mackey and Pearson et al. samplers provided good qualitative samples except on large stones (32–128mm). The Verollet & Tachet sampler lifted little material. The relative abundance of taxa often differed between samplers. Estimates of the number of invertebrates per m² differed widely between samplers except for the Pearson et al. and Ponar grab which gave similar quantitative results at three sites. Values were very low for the Verollet & Tachet sampler and Naturalist's dredge and always very high for the Mackey sampler which often grossly over-sampled its sampling area. Therefore, these three samplers cannot be considered as quantitative. The relationship between the variances and the means of samples of peas in the tank and most invertebrates in the field followed a power law with values of the exponent b in the range 1.13–2.51. The present study completes the evaluation of 14 samplers used to sample benthic macro-invertebrates in deep rivers, and the more important conclusions from the comparative studies of seven grabs, four dredges and three air-lift samplers are summarized in a table.