Growth of Crithidia at High Temperature: Crithidia hutneri sp. n. and Crithidia luciliae thermophila s. sp. n.*

SYNOPSIS. Crithidia hutneri sp. n. and Crithidia luciliae thermophila s. sp. n. are described. Both flagellates can be grown in a defined medium over a temperature range of 15–37°C. The requirements for amino acids, vitamins, purine and hemin, and pH range were similar to those established for Crithidia fasciculata, although threonine was required as a growth factor for C. luciliae thermophila at high temperatures. Adenosine could be used by the 2 Crithidia as a purine source at 28 but not at 37 C.     

First Defined Media for Leptomonas spp. from Insects*

SYNOPSIS. Defined media for Leptomonas collosoma, L. mirabilis, and L. sp. from Dysdercus are described. In addition to factors required by Crithidia spp., including a source of Crithidia factor, all 3 organisms require glycine. Leptomonas collosoma and L. sp. from Dysdercus require choline. All trypanosomatids thus far studied require calcium pantothenate but L. collosoma has an especially large requirement. Leptomonas mirabilis but not L. sp. from Dysdercus can synthesize methionine from homocysteine thiolactone during growth. Homocysteine thiolactone is toxic to L. sp. from Dysdercus. Methionine plus vitamin B₁₂ partly annul homocysteine thiolactone toxicity but only if glycine is also present.     

Enzymes of the Ornithine-Arginine Metabolism of Trypanosomatids of the Genus Crithidia*

SYNOPSIS. Twelve strains of Crithidia, which fall into 8 species, were tested for occurrence of enzymes of ornithine-arginine metabolism. The following enzymes were investigated: arginase, ornithine carbamoyltransferase, argininosuccinate lyase, citrulline hydrolase, arginine deiminase and urease. Arginase and argininosuccinate lyase were found in all species. Citrulline hydrolase was also found in all but the 2 strains carrying endosymbiotes C. deanei and C. oncopelti. On the other hand, ornithine carbamoyltransferase was found only in these 2 strains. Arginine deiminase and urease were absent in all strains. The existence of a common enzymatic pattern for species of the genus Crithidia is thus reported.     

Use of glycoconjugates for trypanosomatid taxonomy

Glycoconjugates from five trypanosomatid genera—Crithidia, Herpetomonas, Endotrypanum, Leishmania, and Trypanosoma—were extracted with Triton X-114 and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by periodic acid-Schiff staining. Most of the glycoconjugates were detected in the hydrophobic phase, indicating the presence of anchored glycoconjugates. All the trypanosomatids expressed a glycoconjugate with a low molecular weigh (below 20 kDa) in this phase. In each species, however, a characteristic and specific pattern of glycoconjugates was also observed in both phases. In the hydrophobic phase: 14–29 kDa lycoconjugates in C. guilhermei; 24–70 kDa in C. fasciculata, C. luciliae, E. schaudinni, and T. cruzi Y and G strains; 45–66 kDa in C. oncopelti and H. samuelpessoai; above 36 kDa in T. dionisii; 20–24 kDa, 36–45 kDa, and 70 kDa in L. tarentolae and T. mega. In the hydrophilic phase, typical glycoproteins were observed in some trypanosomatids: 60 kDa in T. mega and T. cruzi Y strain; 70 kDa in H. samuelpessoai; 66 kDa in C. oncopelti; 20–70 kDa in C. luciliae. These findings suggest that Triton X-114-extracted glycoconjugates could be useful markers for trypanosomatid taxonomy.     

New,known and unidentified species of Eulimdana (Nematoda): additional information on biologically unusual filarioids of charadriiform birds

Eulimdana spp. were found in 13 of 32 species of charadriiform birds from North America and Iceland; 88 of 636 adult birds and seven of 72 juvenile birds harboured infections. Described are: E. andersoni n. sp. from Chlidonias niger; E. metcalforum n. sp. from Sterna hirundo; E. sonini n. sp. from Charadrius hiaticula; E. wongae n. sp. from Limosa fedoa; and E. bainae n. sp. from Numenius phaeopus. Eulimdana asperum Bartlett, Anderson, & Bush, 1989 is reported for the second time in its type-host, Phalaropus tricolor. The emerging pattern of one parasite species to one host species suggests that most Eulimdana species are strongly host specific. However, E. florencae Bartlett & Anderson, 1990 is reported for the first time in a second species of Calidris (C. alba) and may be the unidentified species found in C. bairdii, C. alpina and C. pusilla. Other unidentified species were found in Larus pipixcan, Arenaria interpres and Recurvirostra americana. A key to species-groups (three are proposed) and species of Eulimdana in charadriiforms is given. Two pieces of evidence support an earlier suggestion that infections are acquired by neo-natal charadriiforms: (1) the presence of E. wongae in a bird captured as a chick and examined after two months in captivity; and (2) a greater frequency of mixed sex infections with microfilariae in infected juvenile birds (four of seven) than in infected adult birds (five of 88). The latter also supports an earlier suggestion that post-reproductive worms are ephemeral.     

Species of Calydiscoides Young,l969 (Monogenea: Diplectanidae Bychowsky, 1957: Lamellodiscinae Oliver, 1969) from nemipterid fishes off Peninsular Malaysia

Four new species of Calydiscoides Young, 1969 are described from three species of nemipterids caught off Kemaman, Terengganu, on the eastern coast of Peninsular Malaysia: C. monogrammae n. sp. from Scolopsis monogramma; C. conus n. sp. from S. magaritifer; C. scolopsidis n. sp. from S. margaritifer and S. monogramma; and C. kemamanensis n. sp. from Pentapodus setosus. The present investigation reveals that the squamodiscs (lamellodiscs) are composed of 10-12 short, complete, interlocking and concentric tubular lamellae. The innermost lamella is attached to a pair of adductor muscles.     

Crithidia spp.: Structural comparison of polysaccharides for taxonomic significance

The chemical structures of polysaccharide components of cells of several Crithidia species have been partially elucidated. The structures have been used as criteria to evaluate evolutionary lines previously proposed for species of Crithidia and Herpetomonas and for Trypanosoma cruzi. In accord with the suggestion that Crithidia and Herpetomonas are closely interrelated, all species investigated synthesize a linear (1→2)-linked β-d-mannopyranan and a heteropolysaccharide. These differ from T. cruzi polysaccharide, which contains α-d-mannopyranosyl structures and likely indicates a separate evolutionary route for this flagellate. Crithidia fasciculata, Crithidia harmosae, and Crithidia luciliae form a closely knit group since they form arabinogalactans with related structures. The similarity is particularly close between arabinogalactans of C. fasciculata and C. harmosae whose ¹³C nuclear magnetic resonance spectra show a high degree of resemblance. An unnamed Crithidia sp. contains polysaccharides with fucose and xylose units, intermediate between those of Crithidia deanei, which gave glucose and fucose on hydrolysis, and Herpetomnas samuelpessoai, which gave glucuronic acid and xylose.     

Electrophoretic Analysis of Endonuclease-Generated Fragments of k-DNA,of Esterase Isoenzymes,and of Surface Proteins as Aids for Species Identification of Insect Trypanosomatids1

In order to verify the applicability of biochemical methods for species identification of Trypanosomatidae, 13 species of monoxenic trypanosomatids plus the heteroxenous Trypanosoma cruzi were comparatively analyzed by three different biochemical methods. Insect trypanosomatids examined were: Crithidia acanthocephali, C. fasciculata (three varieties), C. luciliae luciliae, C. luciliae thermophila, C. deanei, C. oncopelti, Herpetomonas muscarum muscarum, H. megaseliae, H. samuelpessoai, H. mariadeanei, Leptomonas seymouri, L. collosoma, L. samueli, and Blastocrithidia culicis. Also included in the survey were aposymbiotic strains of C. deanei and C. oncopelti. Methods used were: electrophoretic profiling of endonuclease-generated fragments of k-DNA, esterase isoenzymes profiling, and polyacrylamide-gel electrophoresis (SDS-PAGE) of radioiodinated cell surface proteins. Interspecific but not intraspecific differences were detected by all three methods among the 13 monoxenic species examined. Thus, it is concluded that these methods can be successfully used, in addition to classical criteria, for species identification of insect trypanosomatids.     

Effect of Temperature and Osmolarity on Growth of Crithidia fasciculata,C. hutneri,C. luciliae thermophila,and Herpetomonas samuelpessoai1

Crithidia fasciculata (Anopheles, Culex, and Nöller strains), C. hutneri, C. luciliae thermophila, and Herpetomonas samuelpessoai were grown in a defined medium with different values of osmolarity at different temperatures. C. fasciculata (all strains) grew best between 300 to 500 mOsm; H. samuelpessoai, 400–500 mOsm; and C. hutneri and C. luciliae thermophila, 500–800 mOsm. At higher temperatures better growth was obtained at the upper osmolarities.     

New and known species of Dactylogyrus Diesing, 1850 (Monogenea,Dactylogyridae) from cyprinid fishes of the River Tigris,Iraq

Four new Dactylogyrus species are described and two redescribed from cyprinids of the River Tigris, Iraq. These are as follows: Dactylogyrus barbioides n. sp. from Barbus grypus; D. orbus n. sp. from Barbus lacerta; D. barbuli n. sp. from Barbus barbulus; D. macrostomi n. sp. from Cyprinion macrostomi; D. pavlovskyi Bychowsky, 1949 from Barbus grypus and Barbus sharpeyi; and D. inutilis Bychowsky, 1949 from Barbus xanthopterus. A phylogenetic and zoogeographical analysis is presented.     

Detection of Candida sp. mannan antigen by indirect ELISA-inhibition

Summary We have obtained mannans from four Candida species: C. albicans A, C. albicans B and C. tropicalis; antimannan sera against C. albicans A, C. albicans B and C. tropicalis were obtained by immunizing rabbits sub-cutaneously with the respective yeast extract. The efficacy of these sera in reacting with mannans obtained from three Candida sp. has been proven by indirect ELISA-inhibition.Any of three immune sera can be used to detect mannan antigen from the three Candida sp. tested. This confirms the existence of crossed reactivity and the possibility of detecting mannan antigen in serum from patients infected by different Candida sp., although we had only one immune serum and one Candida mannan.     

Patterns of parasite infection in bumble bees (Bombus spp.) of Northern Virginia

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Chauhanellus Bychowsky & Nagibina, 1969 (Monogenea) from ariid fishes (Siluriformes) of Peninsular Malaysia

Twelve new species of Chauhanellus Bychowsky & Nagibina, 1969 have been found on six species of ariid from Peninsular Malaysia: Chauhanellus trifidus n. sp., C. digitalis n. sp., C. malayanus n. sp., C. forcipis n. sp. and C. intermedius n. sp. from Arius sagor; C. aspinous n. sp. from Arius venosus; C. caelatus n. sp. from Arius caelatus; C. auriculatum n. sp., C. poculus n. sp. and C. pulutanus n. sp. from Arius maculatus; C. duriensis n. sp. from Arius thalassinus; and C. osteogeneiosi n. sp. from Osteogeneiosus militaris. Some of these Chauhanellus species possess characteristics that are not commonly observed in the genus. C. aspinous n. sp., C. intermedius n. sp. and C. digitalis n. sp. exhibit features found in both Chauhanellus and Hamatopeduncularia: these include absence of spines on the mainpart of the dorsal anchors in C. aspinous n. sp. and C. intermedius n. sp. and presence of haptoral digitation in C. digitalis n. sp. Other features are the five transverse rows of peduncular spines in C. duriensis n. sp., ear-like projections on the anchors in C. auriculatum n. sp., and thin sclerotised plates that partly envelope the ventral anchors in C. forcipis n. sp. Mid-dorsal appendices occur on the dorsal bars of seven of the present species.     

Lepidapedon spp. (Digenea: Lepocreadiidae) from deep-sea gadiform fishes of the NW Atlantic Ocean,including four new species

Five species are described from fishes in deep waters of the northwestern Atlantic Ocean: Lepidapedon gaevskayae n. sp. from Coryphaenoides (Nematonurus) armatus; L. merretti n. sp. from Phycis chesteri; L. zubchenkoi n. sp. from Coryphaenoides (Chalinura) leptolepis; L. beveridgei n. sp. from C. (N.) armatus; and Lepidapedon sp. innom. from Antimora rostrata. The genus Lepidapedon is discussed and divided into groups of species distinguished by the vitelline distribution and the extent of excretory vesicle. The new species are placed in their appropriate groups and differentiated from the other species in the group.     

Neotropical Monogenoidea. 50. Oviparous gyrodactylids from loricariid and pimelodid catfishes in Brazil, with the proposal of Phanerothecioides n. g., Onychogyrodactylus n. g. and Aglaiogyrodactylus n. g. (Polyonchoinea: Gyrodactylidea)

The diagnosis of Phanerothecium Kritsky & Thatcher, 1977 is amended, and Phanerothecioides n. g., Onychogyrodactylus n. g. and Aglaiogyrodactylus n. g., all comprised of oviparous gyrodactylids (Monogenoidea: Polyonchoinea), are proposed to accommodate 11 of the 15 species (14 new to science) herein described and/or reported from loricariid and pimelodid catfishes (Teleostei: Siluriformes) in Brazil: Phanerothecium harrisi Kritsky & Boeger, 1991, P. spinatoides n. sp. and P. deiropedeum n. sp. all from Hypostomus spp.; P. spinulatum n. sp. from Hypostomus cf. regani; Phanerothecioides agostinhoi n. g., n. sp. (type-species) from Hypostomus spp. and Pseudoplatystoma fasciatum; Onychogyrodactylus sudis n. g., n. sp. (type-species) and O. hydaticus n. sp. both from Ancistrus multispinus; Aglaiogyrodactylus forficulatus n. g., n. sp. (type-species) from Kronichthys lacerta; A. pedunculatus n. sp. from Hisonotus sp.; A. guttus n. sp. from Pseudotothyris obtusa; A. salebrosus n. sp., A. conei n. sp. and A. ctenistus n. sp. all from Pareiorhaphis parmula; and A. calamus n. sp. and A. forficuloides n. sp. both from Schizolecis guntheri. Phanerothecioides n. g. is characterised by oviparous forms lacking superficial and deep haptoral bars and pregermarial vitelline follicles, and by having a conspicuous testis in adult specimens, a syncytial prostatic gland, a reduced copulatory sac and vitelline ducts in the form of an inverted ‘U’. Onychogyrodactylus n. g. is differentiated from all other oviparous gyrodactylid genera by its members having a spine-like accessory sclerite enclosed in a separate pouch associated with the terminal male genitalia. Species of Aglaiogyrodactylus n. g. possess H-shaped vitelline ducts and a complex accessory piece and sclerotised or non-sclerotised male copulatory organ enclosed within the copulatory sac.     

Species of Caligus Müller, 1785 (Copepoda: Caligidae) parasitic on marine fishes of Taiwan

Six species of copepods of the genus Caligus (Caligidae, Siphonostomatoida) parasitic on marine fishes of Taiwan are reported. They are: C. absens n. sp. from Priacanthus blochii and P. macracanthus; C. epinepheli Yamaguti, 1936 from Scolopsis vosmeri; C. kanagurta Pillai, 1961 from Decapterus kurroides; C. laticaudus Shiino, 1960 from Lutjanus vitta and Parapristiopoma trilineatum; C. nengai Rangnekar, Rangnekar & Murti, 1953 from Triacanthus biaculeatus; and C. rotundigenitalis Yü, 1933 from Drepane punctata, Liza macrolepis and Terapon jarbua. C. distortus Pillai & Natarajan, 1977 is relegated to a synonym of C. nengai and C. multispinosus Shen, 1957 reported by Lin et al. (1994) from the cultured sea bream Acanthopagrus schlegeli is a misidentification for C. rotundigenitalis Yü, 1933.     

Effects of short‐term exposure to naturally occurring thymol concentrations on transmission of a bumble bee parasite

1. Plants produce antimicrobial phytochemicals that can reduce growth and infectivity of parasites in animals. Pollinator parasites are transmitted between hosts that forage on shared flowers. Floral transmission directly exposes parasites to phytochemicals on floral surfaces and in nectar, both at flowers and, post‐ingestion, in the crop. This exposure could directly affect parasite transmission to new hosts. 2. Nectar chemical analyses were combined with field and cell culture experiments to test the effects of the floral phytochemical thymol on the transmission potential of the trypanosomatid gut parasite Crithidia in Bombus impatiens. First, thymol concentrations in Thymus vulgaris nectar were measured. Second, the effect of adding thymol to floral nectaries on parasite transmission to foraging bees was tested. Third, cell cultures were used to determine direct, dose‐dependent effects of short‐term thymol exposure on subsequent in vitro parasite growth. 3. A total of 26.1 ppm thymol was found in T. vulgaris nectar, five‐fold higher than previously documented in this species. However, addition of thymol to flowers of parasite‐inoculated inflorescences of four plant species did not affect acquisition of Crithidia infection during a foraging bout. Cell culture experiments showed that the thymol concentrations needed to reduce subsequent Crithidia growth by 50% (120 ppm) were 4.6‐fold higher than the highest detected nectar concentration. 4. Although thymol exposure can influence Crithidia viability, Crithidia are robust to the duration and magnitude of exposure encountered during floral foraging under natural conditions. These experiments suggest that any effects of thymol alone on Crithidia–host infection dynamics probably reflect indirect, possibly host‐mediated, effects of chronic thymol ingestion.     

The Lepocreadiidae (Digenea) of pomacentrid fishes (Perciformes) from Heron Island,Queensland, Australia

The following lepocreadiid species are described from pomacentrid fishes from the Southern Great Barrier Reef at Heron Island, Queensland: Lepocreadium adlardi n. sp. from Abudefduf bengalensis; L. clavatum from Acanthochromis polyacanthus and Parma polylepis; Lepocreadium sp. from Amblyglyphidodon curacao; Lepocreadium sp. from Pomacentrus cf. wardi; Preptetos xesuri (new synonyms: P. caballeroi, P. pritchardae Toman, 1989 nec Ahmad, 1984) from Parma polylepis plus the acanthurid Naso annulatus; and P. cannoni from Pomacentrus bankanensis.     

Description of four novel species of Xenorhabdus, family Enterobacteriaceae: Xenorhabdus budapestensis sp. nov., Xenorhabdus ehlersii sp. nov., Xenorhabdus innexi sp. nov., and Xenorhabdus szentirmaii sp. nov

The taxonomic affiliation was determined for four Xenorhabdus strains isolated from four Steinernema hosts from different countries. As compared to the five validly described Xenorhabdus species, i.e., X. nematophila, X. japonica, X. beddingii, X. bovienii and X. poinarii, these isolates represented novel species on the basis of 16S rRNA gene sequences and riboprint patterns, as well as by physiological and metabolic properties. They were named Xenorhabdus budapestensis sp. nov., type strain DSM 16342^T, isolated from Steinernema bicornutum; Xenorhabdus ehlersii sp. nov., type strain DSM 16337^T, isolated from Steinernema serratum; Xenorhabdus innexi sp. nov., type strain DSM 16336^T isolated from Steinernema scapterisci; and Xenorhabdus szentirmaii sp. nov., type strain DSM 16338^T, isolated from Steinernema rarum.