Pleistophora oncoperae sp.n. (Protozoa: Microsporida) from Oncopera alboguttata (Lepidoptera: Hepialidae) in Australia

Pleistophora oncoperae sp.n. is described from adults and larvae of Oncopera alboguttata and O. rufobrunnea. The main site of infection was muscle, though fat body and connective tissue were also infected. Fresh pansporoblasts measured about 25 μm in diameter and contained 16 to 32 or more spores with a mean size of 5.9 × 3.1 μm. Macrospores measuring 7.7 × 4.4 μm were also seen. The mean polar filament length was 158 μm; ultrastructural studies showed that the filament is normally arranged in 14 coils (range, 13 to 20) at an angle of 53.5° to the axis of the spore. The species was found to be distinct from all previously described Pleistophora reported from Lepidoptera.     

Effects of temperature and dosage on Vairimorpha sp. 696 spore morphometrics,spore yield,and tissue specificity in Heliothis virescens

The effects of temperature and dosage on a new microsporidian species, Vairimorpha sp. 696, were examined in H. virescens. The pathogen was evaluated for tissue specificity, spore size, cumulative percentage mortality, and spore production. All tissues examined bore infection at 32°C. Spore length was significantly longer at 19°C (5.9 μm) than at 32°C (4.7 μm). Spore widths at these two temperatures did not differ significantly. Octospores were not found at either temperature at 8 or 12 days postinoculation. One hundred percent mortality was attained in all dosages administered, but the initial rate of mortaily was more rapid in the higher dosages. Finally, spore yield was greater in larvae administered lower dosages. Maximum spore yield at 27°C was 4.87 × 10⁹ spores/larva.     

A novel milky disease organism from Australian scarabaeids: Field occurrence,isolation, and infectivity

A novel milky disease organism has been found causing disease in Aphodius tasmaniae and other scarabaeid larvae in the field in Australia. The sporangium is exceptionally long, measuring 10.5 × 1.5 μm, with a small central spore, measuring 1.0 × 0.6 μm. The vegetative cell is about half the size of the sporangium. The disease was easily transmitted by injection of spores into the hemocoel, with typically milky symptoms developing in 2–4 weeks. Spores will form in vivo at temperatures down to 12°C. For A. tasmaniae third-instar larvae, the ID₅₀ by injection was 3 × 10² spores/larva, yet no infection resulted when larvae were reared in peat containing up to 10⁸ spores/g, i.e., the disease was not successfully transmitted per os. All 10 species of scarabaeids tested were susceptible to the disease when spores were injected; however, all attempts to infect larvae per os were unsuccessful. In vitro culture was also unsuccessful.     

A Nosema species of the Egyptian cotton leafworm, Spodoptera litura (Lepidoptera): Its morphology,development, host range,and taxonomy

A microsporidian was isolated from the Egyptian cotton leafworm, Spodoptera litura, and was identified as a member of the genus Nosema since its sporont produced two spores. The size of the fresh spores was 3.3–4.5 × 1.7 – 2.2 μm. Mean length of the polar filaments was 90.7 μm. In addition to the Egyptian cotton leafworm, this microsporidian was also infectious to Pieris rapae crucivora, Hyphantria cunea, Chilo suppressalis, Agrotis ipsilon, and Adoxophyes orana. Although the parasite ultimately caused generalized infection in the Egyptian cotton leafworm larva, the main site of infection was the adipose tissue.     

Effect of Temperature on Fecundity,Life Span and Morphology of Long‐ and Short‐Spined Clones of Brachionus caudatus f. apsteini (Rotifera)

We investigated the effect of temperature (20, 25 and 30 °C) on fecundity, life span and morphology of the rotifer Brachionus caudatus f. apsteini. For each temperature, short posterior‐spined and long posterior‐spined clones of B. caudatus f. apsteini were individually cultured for up to six generations. The rotifers were fed Chlorella sp. at a density of 1 × 10⁶ cells ml^–1. Morphometric data (body size and spine length) were collected. Total number of offspring producing by a single female per life cycle at high temperature was higher than at low temperature. The duration of juvenile period, reproductive period, post‐reproductive period and life span of both clones of B. caudatus f. apsteini decreased with increasing temperature. All offspring of short posterior‐spined clone produce posterior spines at 20 and 25 °C, with an average length of 19.8 ± 6.6 and 11.9 ± 2.6 μm, respectively. In contrast, they cannot develop posterior spines at 30 °C, at which the average length of the posterior spine remnant was 6.4 ± 1.3 μm. On the other hand, all offspring of long posterior‐spined clone have long posterior spines with average lengths of 36.8 ± 6.1, 36.3 ± 5.2 and 36.6 ± 6.2 μm at 20, 25 and 30 °C, respectively. This study indicated that the production of posterior spines can be induced by low temperature and that short posterior‐spined and long posterior‐spined clones are genetically different. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Using KCl to determine size at competence for larvae of the marine gastropod Crepidula fornicata (L.)

Competent larvae of the marine gastropod Crepidula fornicata (L.) were induced to metamorphose (i.e., lose the velum) by elevating sea-water [KCl] by 5–50 mM. The response was optimal at 15–20-mM elevations, at which 50% metamorphosis was obtained in <4 h. Larvae that did not metamorphose during brief exposures (1–5 h) to elevated [KCl] generally maintained the larval form following transfer to control sea water, suggesting that competent larvae must be continuously immersed in the test solutions for metamorphosis to occur. The smallest larvae to respond to elevated [KCl] had shell lengths of ≈700–800 μm, the range of shell lengths within which larvae of this species become responsive to natural inducers. All larvae >≈1125 μm shell length metamorphosed in response to increased [KCl]. Rearing temperature may affect the size at which larvae of this species become responsive to K⁺. CaCl₂ (20-mM concentration elevations), GABA (4×10⁻⁷, 4×10⁻⁶ M), and NaCl (10–20-mM concentration elevations) generally failed to trigger metamorphosis. Twenty-mM elevations of [RbCl] and [CsCl] induced 100% metamorphosis but the juveniles were immobile and died after several days. Elevating [KCl] appears to be a reliable way to assess competence and trigger metamorphosis in larvae of C. fornicata.     

The susceptibility ofTribolium castaneum [Col.: Tenchrionidae] toFarinocystis tribolii [Protozoa: Schizogregarinida]

The LT₅₀ ofFarinocystis tribolii Weiser to larvae ofTribolium castaneum (Herbst) increased with the age of the insect indicating that older larvae were relatively more tolerant to the infection though there was 100 % mortality ultimately. The adults were less susceptible than larvae and between sexes, females were more susceptible than males. The number of spores produced increased with the stage of the larvae, but there was no variation in the size of spores in the different instars. The LC₅₀ on 20th and 40th day of inoculation were 1.4×10⁷ and 2.1×10⁶ respectively. Mortality-time due toF. tribolii was shorter at 35 °C than at 25 °C. Sporulation occurred earlier at 35 °C than at 25 °C.     

Coccidian Parasites (Apicomplexa: Eimeriidae) from Insectivores. IV. Four New Species in Talpa europaea from England1

ABSTRACT. Moles from England were examined for coccidian oocysts and all 64 Talpa europaea were infected; of 64 infected hosts, 56 (88%) had multiple infections representing two to six coccidian species when examined. Oocysts in 31 of the 64 samples remained unsporulated. Three eimerians and one isosporan were studied from the 33 fecal samples that had sporulated oocysts and these are described as new species; Cyclospora talpae Pellérdy & Tanyi, 1968, and Isospora sofiae (Golemansky, 1978) Levine & Ivens, 1979, are redescribed; and Cyclospora sp., similar to C. talpae, is discussed. Sporulated oocysts of C. talpae are ellipsoidal, 14.3 × 9.6 (12–19 × 6–13) μm with sporocysts ovoid, 9.4 × 5.7 (6–13 × 4–8) μm; it was found in 21 of the 33 (63.6%) sporulated samples. Sporulated oocysts of Cyclospora sp. are subspheroidal to ellipsoidal, 12.5 × 8.9 (10–14 × 6–12) μm with sporocysts ovoid, 8.6 × 5.3 (6–10 × 4–6) μm; it was found in 21 of the 33 (63.6%) sporulated samples. Sporulated oocysts of Eimeria avonensis n. sp. are elongate-ellipsoidal, 15.0 × 9.6 (13–20 × 7–12) μm with sporocysts ovoid, 6.6 × 3.6 (5–9 × 3–7) μm; it was found in 15 of the 33 (45.5%) sporulated samples. Sporulated oocysts of Eimeria berea n. sp. are subspheroidal, 12.1 × 10.5 (10–15 × 8–14) μm with sporocysts ovoid, 6.3 × 3.9 (5–10 × 2–5) μm; it was found in 8 of the 33 (24.2%) sporulated samples. Sporulated oocysts of Eimeria globula n. sp. are spheroidal, 20.9 × 19.9 (19–24 × 17–21) μm with sporocysts elongate-ovoid, 11.5 × 6.9 (9–16 × 6–9) μm; it was found in 3 of the 33 (9.1%) sporulated samples. Sporulated oocysts of Isospora sporopointaea n. sp. are subellipsoidal to ellipsoidal, 17.1 × 11.4 (13–21 × 8–14) μm with sporocysts ellipsoidal with both ends pointed, 11.9 × 5.9 (9–16 × 4–8) μm; it was found in 27 of the 33 (81.8%) sporulated samples. Sporulated oocysts of I. sofiae are spheroidal to subspheroidal, 12.2 × 11.0 (9–16 × 8–15) μm with sporocysts ovoid, 9.1 × 5.2 (6–13 × 3–8) μm; it was found in 25 of the 33 (75.8%) sporulated samples. To date, the coccidian parasites of talpids include two cyclosporans, 12 eimerians, and six isosporans, exclusive of the four new species described here.     

The morphology and development of Nosema carpocapsae,a microsporidian pathogen of the codling moth,Cydia pomonella (Lepidoptera: Tortricidae) in New Zealand

The morphology of Nosema carpocapsae and its development in experimentally infected codling moth larvae are described. Spherical uninucleate meronts were the first stages. Nuclear division produced binucleate meronts which were the most abundant vegetative stage, although additional uninucleate and a few tetranucleate meronts were also observed at this time. All meronts were spherical and ranged from 2.8 to 5.8 μm in diameter. Uninucleate and binucleate fusiform sporonts then appeared followed by some tetranucleate and dividing forms. Oval sporoblasts developed after these and did not divide before maturing into spores. Sporonts were approximately 5.0 to 7.9 × 2.4 to 3.0 μm. Spores developed in all host tissues except the nervous tissue. The binucleate spores showed considerable variation in spore size, 2.4 to 3.9 × 1.3 to 3.1 μm (alcohol fixed, Giemsa stained). The polar filament was usually coiled 11 times (range 9 to 13) at an angle of 53° to the long axis of the spore. Its maximum observed length was 75 μm.     

Physiological and biochemical energetics of larvae of Teredo navalis L. and Bankia gouldi (Bartsch) (Bivalvia : Teredinidae)

The biochemical composition of larvae of Teredo navalis L. and Bankia gouldi (Bartsch) (Bivalvia: Teredinidae) was examined throughout larval development at 23°C and 30–32%. salinity in the presence of the phytoplankton food Isochrysis aff. galbana (clone T-ISO), during a delay of metamorphosis in the presence of food but absence of a wood substratum and during periods of enforced starvation. Newly released Teredo navalis larvae had a mean length (L) and height (H) of 89.3 and 76.1 μm respectively. The first appearance of pediveliger larvae at 212.1 μm L and 230.0 μm H occurred 27 days after release. Larval dry weight increased from 0.29 μg to 1.96 μg during this period. Newly formed straight hinge larvae of Bankiagouldi had dimensions of 62.8 μm L and 49.8 μm H. Metamorphically competent B. gouldi larvae had dimensions of 230.0 μm L and 282.9 μm H and were first observed 20 days after fertilization. Larval dry weight increased from 0.06 μg to 2.20 μg during this period. During enforced delay of metamorphosis the ash-free dry weight of Teredo navalis larvae decreased whereas the ash free dry weight of Bankia gouldi larvae increased. During the early period of shelled larval development both species showed similar decreases in lipid, protein and carbohydrate levels (μg·mg dry weight^?1); however, this was reflected in a decrease in biochemical content (μg·larva^?1) only in Teredo navalis. During enforced starvation the major proportion of both the weight and caloric losses were due to protein. Lipid also contributes significantly to these losses whereas the contribution of carbohydrate was small. Larval oxygen consumption rates were determined directly by manometry and indirectly by estimates of decrease in caloric content during periods of enforced starvation. Direct and indirect determinations for T. navalis are described by the relationships R = 1.16 W^1.05 and R = 0.98 W^1.24 respectively where R is the respiration rate in nl O₂ · larva^?1 · h^?1 and W is dry weight inclusive of shell in μg. Direct and indirect determinations for Bankia gouldi are described by the relationships R = 1.37 W^1.25 and R = 1.81 W^1.25 respectively. When data for both assay procedures are combined for each species the relationships R = 1.10 W^1.07 and R = 1.44 W^1.18 are obtained for Teredonavalis and Bankia gouldi respectively.     

Coccidian Parasites (Apicomplexa: Eimeriidae) from Insectivores. VI. Six New Species from the Eastern Mole,Scalopus aquaticus1

ABSTRACT Thirteen eastern moles, Scalopus aquaticus, collected in West Texas were examined for coccidian oocysts; 11 (85%) were infected and eight (73%) of these had multiple infections representing two or more species. One cyclosporan, three eimerians, and two isosporans were studied and all are described as new species. Sporulated oocysts of Cyclospora megacephali n. sp. were subspheroidal, 18.5 × 15.7 (14–21 × 12–18) μm; they had sporocysts pointed at one end with Stieda bodies nearly as wide as the sporocysts themselves, and were 15.0 × 7.2 (11–17 × 6–9) μm; C. megacephali was found in four (31%) hosts. Sporulated oocysts of Eimeria scalopi n. sp. were spheroidal to subspheroidal, 13.6 × 12.6 (11–17 × 11–15) μm with sporocysts lemon-shaped, 8.7 × 5.5 (7–10 × 4–7) μm; it was found in six (46%) hosts. Sporulated oocysts of Eimeria aquatici n. sp. were asymmetrically ellipsoidal, 17.0 × 10.6 (14–20 × 9–14) μm, with sporocysts elongately ovoidal, 9.0 × 5.2 (8–11 × 4–6) μm; it was found in two (15%) hosts. Sporulated oocysts of Eimeria motleiensis n. sp. were subspheroidal, 17.0 × 15.3 (15–20 × 13–18) μm with sporocysts ovoidal, 10.7 × 6.8 (10–13 × 6–8) μm; it was found in seven (54%) hosts. Sporulated oocysts of Isospora motleiensis n. sp. were spheroidal to subspheroidal, 13.6 × 12.0 (10–17 × 8–15) μm with sporocysts broadly ovoidal, 9.5 × 6.7 (7–11 × 4–8) μm; it was found in nine (69%) hosts. Sporulated oocysts of Isospora aquatici n. sp. were subspheroidal, 20.9 × 18.4 (15–24 × 13–21) μm with sporocysts ellipsoidal, 11.8 × 9.0 (9–14 × 7–11) μm; it was found in two (15%) hosts.     

Schistosoma mansoni: Post-transformational surface changes in schistosomula grown in vitro and in mice

The development of schistosomula during the first 4 days after transformation from cercariae has been examined in parasites isolated from the lungs of mice and in organisms cultured in lactalbumin and rabbit serum or in the defined serum-free medium, RPMI 1640. The development of organisms grown under all three conditions was the same. Schistosomula increased in length from 67 to 110 μm and decreased in width from 24 to 18 μm, so that the volume remained constant at approximately 2.7 × 10⁴ μm³. The increase in length occurred mainly in the torso or posterior three-quarters of the worm which increased from 49 to 88 μm or 80%, whereas the head increased from 18 to 22 μm or 22%. The spines were lost from the surface that was most rapidly lengthening by gradual resorption into the tegument and were replaced by pits mainly during the first 3 days. These changes resulted in a 325% increase in the surface area of the schistosomula, from 1.2 × 10⁴ to 3.9 × 10⁴ μm². In addition, the openings of the acetabular ducts, the ventral sucker, and the tail socket all became smaller and flatter over the four-day period. Internally, the major changes were the loss of the acetabular ducts in the pre- and post-acetabular glands and an increase in size of the caecum. In summary, these experiments show that the surface of the schistosomulum is extensively remodeled before intravascular migration occurs and demonstrate the efficacy of RPMI 1640 as a culture medium for schistosomula in the first 4 days after transformation.     

<Emphasis Type="Italic">Myxobolus imparfinis</Emphasis> n. sp. (Myxozoa: Myxosporea), a new gill parasite of <Emphasis Type="Italic">Imparfinis mirini</Emphasis> Haseman (Siluriformes: Heptapteridae) in Brazil

A new species of myxozoan, Myxobolus imparfinis n. sp. is described based on material from the gills of Imparfinis mirini (Haseman) (Heptapteridae). Mature myxospores are round, measuring 7.1–8.4 (7.9 ± 0.3) μm in length, 4.5–6.2 (5.5 ± 0.5) μm in width and 3.1–4.2 (3.7 ± 0.3) μm in thickness. The polar capsules are of unequal size, the larger polar capsule measuring 3.4–4.5 (3.9 ± 0.3) μm in length and 1.4–2.0 (1.7 ± 0.1) μm in width and the smaller capsule measuring 3.1–3.8 (3.4 ± 0.2) μm in length and 1.2–1.8 (1.5 ± 0.2) μm in width. The polar filament presents 6–7 coils. Spores had a prevalence of infection of 75% (6/8). In histological analyses we detected the development site of spores in primary filaments, in afferent branchial artery, thus classifying the type of infection to the filamental type and vascular subtype. The phylogenetic analyses of a dataset including species Myxobolus Bütschli, 1882 and Henneguya Thélohan, 1892 from South America recovered M. imparfinis n. sp. as a sister species of Myxobolus flavus Carriero, Adriano, Silva, Ceccarelli & Maia, 2013. To our knowledge, this is the first record of a myxozoan species parasitising I. mirini.     

Coccidian Parasites (Apicomplexa: Eimeriidae) from Insectivores. II. Six New Species from Japanese Shrew Moles (Talpidae)1

ABSTRACT. Thirty-eight of 51 (74.5%) shrew moles collected in Japan were infected with from one to four species of Eimeria and/or Isospora including six of six Dymecodon pilirostris and 32 of 45 (71.1%) Urotrichus talpoides. Four eimerians and two isosporans were identified and all are described as new species. Sporulated oocysts of Eimeria amorphospora n. sp. were subspheroid/ellipsoid, 21.1 × 17.9(18-25 × 16-21) μm. Sporocysts were amorphous, gelatinoid envelopes 20.3 × 7.5 (17–24 × 7–9) μm. Sporozoites were enclosed together within a membrane in each sporocyst. This species was found in 9 of 45 (20%) U. talpoides. Sporulated oocysts of Eimeria gonocilia n. sp. were subspheroid/ellipsoid, 28.8 × 24.4 (25–30 × 21–28) μm; a highly ornate outer oocyst wall gave the appearance of a ciliated ball. Sporocysts ovoid, pointed at both ends, were 17.0 × 9.0 (15–19 × 7–11) μm; this species was found in 4 of 45 (8.9%) U. talpoides. Sporulated oocysts of Eimeria talpoidei n. sp. were asymmetrical ovoid, 20.6 × 13.3 (18–23 × 12–15) μm, with sporocysts lacrimiform, 12.0 × 5.8 (10–14 × 5–7) μm. This species was found in 7 of 45 (15.6%) U. talpoides. Sporulated oocysts of Eimeria honshuensis n. sp. were ellipsoid, 15.5 × 11.4 (13–18 × 10–13) μm, with sporocysts ovoid, 9.1 × 5.2 (8–10 × 4–6) μm. This species was found in 10 of 45 (22.2%) U. talpoides and in 5 of 6 (83.3%) D. pilirostris. Sporulated oocysts of Isospora dymecodi n. sp. were subspheroid/ellipsoid, 15.8 × 12.6 (13–17 × 11–13) μm, with sporocysts ellipsoid, 10.9 × 6.9 (10–13 × 6–8). This species was found in six of six D. pilirostris. Sporulated oocysts of Isospora urotrichi n. sp. were spheroid/subspheroid, 13.4 × 12.4 (11–16 × 9–14) μm, with sporocysts ovoid, 9.2 × 6.3 (8–11 × 5–7) μm. This species was found in 27 of 45 (60%) U. talpoides. Only 14 of 38 (36.8%) infected hosts (one D. pilirostris, 13 U. talpoides) were seen to be naturally infected with only one coccidian species when sampled.     

Coccidia of Brazilian Mammals: Eimeria corticulata N. Sp. (Apicomplexa: Eimeriidae) from the Anteater Tamandua tetradactyla (Xenarthra: Myrmecophagidae) and Eimeria zygodontomyis N. Sp. from the Cane Mouse Zygodontomys lasiurus (Rodentia: Cricetidae)

Feces from a specimen of Tamandua tetradactyla (Linn.) from Portel, Para State, north Brazil, contained two different coccidial oocysts; one identified as Eimeria tamanduae Lainson 1968, and the other as a new species, described here as Eimeria corticulata n. sp. Oocysts of E. corticulata are ellipsoidal, 37.4 × 30.4 (31.2–43.7 × 23.7–35.0) μm, shape index (length/width) 1.2 (1.0–1.5). Oocyst wall 2.5–3.7 μm thick and composed of two layers; an outer thick, brown-yellow one with radial striations, and a thin inner smooth one: no visible micropyle. Oocyst residuum a large globule of about 10.7 × 10.3 μm, usually accompanied by a number of smaller attached globules. Sporocysts ellipsoidal, 21.0 × 11.0 (20.0–22.5 × 10.0–12.5) μm, with a conspicuous Stieda body: shape index 1.9 (1.6–2.2). Sporocyst residuum a small number of scattered granules: sporozoites 18.7 × 5.0 μm, with a large posterior refractile body. Eimeria zygodontomyis n. sp. is described in feces from Zygodontomys lasiurus (Lund) from the Serra dos Carajas, Para. Oocysts ellipsoidal to cylindrical, 16.5 × 12.0 (13.7–18.7 × 11.2–12.3) μm, shape index 1.4 (1.2–1.5). Wall colorless, smooth, single-layered and about 0.6 μm thick: no micropyle. No oocyst residuum, but a polar granule of about 1.8 × 1.0 μm is sometimes present. Sporocysts ellipsoidal, 8.4 × 5.5(7.5–8.7 × 5.0–6.2) μm, shape index 1.5 (1.4–1.7), with a thin colorless wall and a delicate Stieda body. Sporozoites enclose a compact residuum of about 2.5 × 3.7 μm.     

Four new coccidia (Apicomplexa: Eimeriidae) from the Plateau zokor,Myospalax baileyi Thomas (Rodentia: Myospalacinae), a subterranean rodent from Haibei area,Qinghai Province,China

Thirty-eight faecal samples from the Plateau zokor, Myospalax baileyi Thomas, collected in the Haibei Area, Qinghai Province, China, were examined for the presence of coccidia (Apicomplexa: Eimeriidae). Seventeen of 38 faecal samples (44.7%) were found to contain coccidian oöcysts representing four new species of Eimeria Schneider, 1875, and four of 17 (23.5%) infected zokors were concurrently infected with two or three of these eimerian species. The sporulated oöcysts of Eimeria myospalacensis n. sp. are ovoidal, 9.5–17.0 × 8.0–13.0 (mean 13.0 × 10.4) μm; a polar granule is present, oöcyst residuum is absent; sporocysts are ovoidal, 4.5–7.5 × 3.0–5.0 (mean 6.3 × 4.2) μm and have both a Stieda body and residuum. Oöcysts of Eimeria fani n. sp. are ellipsoidal to cylindroidal, 12.5–16.0 × 8.0–11.0 (mean 14.6 × 9.9) μm; a polar granule is present, but micropyle and residuum are lacking; sporocysts are ovoidal, 4.5–7.5 × 3.0–5.3 (mean 6.7 × 4.4) μm; a residuum and a Steida body are present. Oöcysts of Eimeria baileyii n. sp. are ellipsoidal, 15.0–23.0 × 12.0–18.0 (mean 18.2 × 13.7) μm; a polar granule is present but oöcyst residuum is absent; sporocysts are ovoidal, 8.0–11.0 × 5.0–7.0 (mean 9.5 × 5.9) μm and have both a Stieda body and residuum. Oöcysts of Eimeria menyuanensis n. sp. are ovoidal, 12.5–21.0 × 11.0–18.0 (mean 17.1 × 14.6) μm, with a distinct micropyle c.2.5 μm wide; a polar granule is present but a residuum is absent; sporocysts are ovoidal, 8.0–12.0 × 5.0–7.0 (mean 10.2 × 6.4) μm, and have both a Stieda body and residuum.     

Rugopharynx rosemariae new species (nematoda: Pharyngostrongylidae) from grey kangaroos (Macropus gig anteus and M. fuliginosus) with life cycle stages and associated pathology

Beverdge I. and Ppresidente P. J. A. 1978. Rugopharynx rosemarie sp. nov. (Nematoda: Pharyngostrongylidae) from grey kangaroos (Macropus giganteus and M. fuliginosus) with life cycle stages and associated pathology. International Journal for Parasitology8: 379–387. Rugopharynx rosemariae new species is described from the stomachs of grey kangaroos, Macropus giganteus Shaw, 1790 and Macropus fuliginosus (Desmarest, 1817) from south-eastern Australia. The new species differs from other species of the genus in spicule length and in the length and shape of the oesophagus. Parasitic life-cycle stages are described from natural infections; free-living life-cycle stages were obtained by culturing eggs from gravid females. Third stage larvae burrow into the gastric mucosa producing small elevated nodules resulting from a localized fibroplastic and inflammatory reaction in the lamina propria and submucosa. Fourth stage and adult worms occur in the stomach lumen. Rugopharynx brevis (Canavan, 1931) is made a synonym of Rugopharynx australis (Mönnig, 1926).     

A new species of Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) from the dusky rice rat Melanomys caliginosus (Tomes) (Mammalia: Rodentia) in Costa Rica

We collected faecal samples from 24 dusky rice rats, Melanomys caliginosus (Tomes) (Rodentia: Cricetidae: Sigmodontinae), in a Biological Reserve in Costa Rica, and found three (12.5%) to be infected with a species of Eimeria Schneider, 1875, which we describe here as new. Sporulated oöcysts of Eimeria caliginosa n. sp. are almost spheroidal and measure 16–21 × 17–20 (mean 19.6 × 18.2) μm; micropyle, oöcyst residuum and polar granule are absent. Sporocysts are ovoidal, 9–13 × 6–8 (mean 11.2 × 6.7) μm, with small Stieda and sub-Stieda bodies present, but a para-Stieda body is absent; the sporocyst residuum is a compact mass of c.11–15 granules, c.5 μm wide. Sporozoites are crescent-shaped, 5–8 × 2–3 (mean 6.8 × 2.4) μm. This is the third species of Eimeria described from the genus Melanomys Thomas.     

Ligation of EcoRI endonuclease-generated DNA fragments into linear and circular structures.

Double-stranded DNA fragments terminated at their 5′-ends by the singlestranded sequence pA-A-T-T-, generated by digestion of DNA with EcoRI restriction endonuclease, were ligated with Escherichia coli polynucleotide ligase under various conditions of temperature, concentration and time. The linear and circular products of ligation were separated by electrophoresis in agarose gel and quantitated by densitometry. The rate of ligation of (EcoRI-cleaved) simian virus (SV40) DNA at a concentration of 100 μg/ml increased from 0 °C to 5 °C to 10 °C (6-fold increase overall); raising the temperature to 15 °C did not further increase the rate of ligation. At the appropriate DNA concentrations, the predominant products of ligation are either linear concatemers that are integral multimers of the starting DNA fragment, or covalently closed circular structures of the monomeric DNA fragment. Ligating a mixture of two different length DNA fragments gives rise to all of the possible expected recombinant molecules.Linear or circular products of ligation were predicted by consideration of the total concentration of DNA termini, i, and the local concentration of one terminus in the neighborhood of the other on the same DNA molecule, j. The parameter j is a function of the length of a DNA molecule, providing this length is greater than the random coil segment of DNA. Experimentally it was found that circular structures are formed in significant amounts only under conditions when the value of j is several times greater than that of i. When j = i, equal amounts of linear and circular products would be expected, but most of the molecules were ligated into linear concatemers. No circular structure of a DNA fragment whose contour length l (6 × 10⁻² μm) is smaller than the random coil segment value b (7·17 × 10⁻² μm) was observed, while circular structures of the dimer of the same molecule (12 × 10⁻² μm) were detected.     

Cryptosporidium proventriculi sp. n. (Apicomplexa: Cryptosporidiidae) in Psittaciformes birds

Cryptosporidiosis is a common parasitic infection in birds that is caused by more than 25 Cryptosporidium species and genotypes. Many of the genotypes that cause avian cryptosporidiosis are poorly characterized. The genetic and biological characteristics of avian genotype III are described here and these data support the establishment of a new species, Cryptosporidium proventriculi. Faecal samples from the orders Passeriformes and Psittaciformes were screened for the presence of Cryptosporidium by microscopy and sequencing, and infections were detected in 10 of 98 Passeriformes and in 27 of 402 Psittaciformes. Cryptosporidium baileyi was detected in both orders. Cryptosporidium galli and avian genotype I were found in Passeriformes, and C. avium and C. proventriculi were found in Psittaciformes. Cryptosporidium proventriculi was infectious for cockatiels under experimental conditions, with a prepatent period of six days post-infection (DPI), but not for budgerigars, chickens or SCID mice. Experimentally infected cockatiels shed oocysts more than 30 DPI, with an infection intensity ranging from 4,000 to 60,000 oocysts per gram (OPG). Naturally infected cockatiels shed oocysts with an infection intensity ranging from 2,000 to 30,000 OPG. Cryptosporidium proventriculi infects the proventriculus and ventriculus, and oocysts measure 7.4 × 5.8 μm. None of the birds infected C. proventriculi developed clinical signs.