An unusual intraerythrocytic parasite of Parablennius cornutus from South Africa

An intertidal horned blenny, Parablennius cornutus, captured at De Hoop Nature Reserve, South Africa, was found to harbor an unusual blood parasite and the haemogregarine Haemogregarina bigemina. In Giemsa-stained blood films, the enigmatic parasite occurred primarily as intraerythrocytic ringlike stages, with unstained centers and peripheral bands of beaded chromatin, not unlike Haemohormidium spp. Larger forms of the same organism stained pink with Giemsa, with nuclei occurring as 4-8 minute structures around the parasite body or distributed within it. These larger parasites apparently segmented into up to 8 individuals that were rounded or oval with deep-stained, comma-shaped or polar regions surrounding blue cytoplasm. Extracellular, binucleate, sporelike structures in clusters of as many as 16 individuals were also seen in blood films. Praniza larvae of the isopod Gnathia africana were seen in histological sections of gill tissue. Examination of spleen tissue by transmission electron microscopy showed intraerythrocytic organisms with ultrastructural characteristics like those of Haematractidium scombri, namely, a single boundary membrane, sometimes closely apposed nuclei with nucleoli, and profiles of dense material of variable structure. It is concluded that the parasite is probably related to Haemohormidium spp. and H. scombri, but it also shares features with some Microsporida.     

Size differences in hamuli of Kuhnia scombri (Monogenea: Polyopisthocotylea) from different geographical areas not due to differences in host size

Hamuli of Kuhnia scombri from Scomber australasicus and S. japonicus from several localities in the Pacific Ocean show marked differences in length. Hamuli from large fish are slightly longer than those from small fish caught in the same geographical area (northern New Zealand). Differences correlated with host size are much less than those associated with geographical area and may be due to infections acquired in different localities. Taxonomic studies of Monogenea must take geographical variation and, to a lesser degree and in at least some species, effects of host size into consideration.     

Gill monogeneans of the chub mackerel, Scomber japonicus from Madeiran waters of the Atlantic Ocean, Portugal

Five species of monogeneans were recovered from the gill filaments of 181 chub mackerel, Scomber japonicus, from the Madeiran waters of the Atlantic Ocean, Portugal, during 2004/2005. The monogenean Pseudokuhnia minor showed the highest prevalence (98.68%) and a mean intensity of 28.23, followed by Kuhnia scombri (prevalence of 43.71% and mean intensity of 2.69) and K. scombercolias (prevalence of 39.1% and a mean intensity of 1.81). Kuhnia sprostonae and Grubea cochlear were rare, occurring in only one and five fish hosts respectively. No correlation between fish host length and mean intensity of infection with the three most abundant monogeneans was found. However, significant differences in prevalence and abundance were found in relation to date of sampling for P. minor, and all parasites were aggregated in their distribution.     

Different populations of Scomber australasicus in New Zealand and south-eastern Australia, demonstrated by a simple method using monogenean sclerites

A simple method for distinguishing fish populations using monogenean sclerites is described. Monogeneans are left in a drop of 45% acetic acid on a microscope slide for a few minutes, and squashed under a cover slip to spread the hamuli and genital hooks in one plane. Hamuli and hooks are then measured with the aid of measuring eyepieces. Measurements of sclerites of Kuhnia and Pseudokuhnia fixed directly in 10% formalin or fixed in 10% formalin after freezing for several days gave the same results. Prerequisites for the use of the method are that monogeneans occur in sufficient numbers and show distinct geographical variation. The method is easiest to use if the size of sclerites does not change with body growth. Kuhnia scombri (Monogenea: Polyopisthocotylea) from the mackerel, Scomber australasicus , in New Zealand and New South Wales differ significantly in the length of the hamuli; it is concluded that there may be different populations of mackerel at the two localities. There also are different populations of Scomber japonicus in Japan and Ecuador, and sympatric populations of S. japonicus and S. australasicus in Japan harbour identical populations of the monogeneans Pseudokuhnia minor and Kuhnia sprostonae.     

Mitochondrial DNA diversity in North American and European Atlantic salmon with emphasis on the Downeast rivers of Maine

The displacement loop and NADH-1 dehydrogenase regions of mitochondrial DNA (mtDNA) were amplified by the polymerase chain reaction in 954 Atlantic salmon and digested with 40 restriction endonucleases. Variation was detected with 10 enzymes, resulting in 21 composite haplotypes which were strongly patterned geographically with a major discontinuity observed between most North American (NA) and European salmon. Significant heterogeneity of haplotype frequencies was found within and among all classification levels (continent, country, and river). Haplotype frequencies were significantly different across continents, within European samples, within NA samples, within Canadian samples, within wild Maine samples, within captive Maine strains, and between captive and wild Maine strains. Nine haplotypes occurred only in NA, seven in Maine, three only in Maine, and 11 occurred only in Europe. Some Maine rivers had only a single haplotype, suggesting that effective population sizes may be low. The second most frequent European haplotype occurred in tributaries to one Newfoundland river. Gene trees based on parsimony and genetic distance suggest that the haplotypes are monophyletic within each continent, and that the haplotype found on both continents is intermediate between those of Europe and NA, suggesting common ancestry of all haplotypes.     

Growth of Bacteroides fragilis group in agar and broth media

The rate of bacterial growth of four Bacteroides fragilis group organisms was determined in agar and broth media. Exponential bacterial growth occurred in agar media within 4 to 8 h, while such growth was delayed in broth media and occurred within 12-24 h after inoculation. This phenomenon may explain why antimicrobials which manifest an 'inoculum effect' may show increased resistance to antimicrobials when tested in agar media.     

Interaction of catenaria anguillulae with romanomermis culicivorax

The development of Catenaria anguillulae, at 27 degrees C, in live and heat-immobilized Romanomermis culicivorax was studied at the ultrastructural level. C. anguillulae zoospores infected eggs, preparasites, and postparasites. The zoospores attached predominately to the head of preparasites but in postparasites attachment was primarily on the attenuated tail region. Zoospore attachment usually occurred within 4 h but at high zoospore densities attachment occurred within 30 min. Infection hyphae penetrated the cuticle and developed sporangial swellings within the nematode tissues and pseudocoelom. Sporangia produced exit tubes within 48 h. Those regions of the J3 and J4 nematodes infected by C. anguillulae became immobile whereas uninfected regions remained mobile. The J2 infected nematodes usually died within 1-2 days. Tissues adjacent to C. anguillulae hyphae or developing sporangia appeared to be relatively unaffected and ultrastructurally normal. The lack of an apparent host response in the nematode suggested either a lack of hypersensitivity or a long evolutionary host-parasite relationship. Copyright 1999 Academic Press.     

Linker mutagenesis in the lacZ gene of Escherichia coli yields variants of active beta-galactosidase

Synthetic octameric oligonucleotides that code for a unique restriction site were cloned into a randomly linearized plasmid that carries the lacZ gene. The insertions were mapped by digestion with appropriate restriction endonucleases. 12 mutants were identified which carry an insertion within the lacZ gene and still express active beta-galactosidase. Small deletions or duplications of the wild-type sequence occurred at these positions which restore the correct reading frame. The insertions occurred in the first and the last third of the internal duplication of the lacZ gene and within the domain homologous to dihydrofolate reductase.     

Kuhnia sprostonae Price, 1961 and K. scombercolias Nasir & Fuentes Zambrano, 1983 (Monogenea: Mazocraeidae) and their microhabitats on the gills of Scomber australasicus (Teleostei: Scombridae), and the geographical distribution of seven species of gill Monogenea of Scomber spp.

As a result of an extensive survey of monogeneans on the gills of scombrid fishes of the genus Scomber, especially S. australasicus in Australia, it has been shown that Kuhnia sprostonae as previously defined by Rohde & Watson is, in fact, two distinct species, namely K. sprostonae Price, 1961 and K. scombercolias Nasir & Fuentes Zambrano, 1983. K. sprostonae inhabits the pseudobranchs of Scomber scombrus in the North Sea, S. japonicus off Madeira and S. australasicus in Australian waters, and shows considerable variability in the lengths of the hamuli and the genital hooks. K. scombercolias occurs on the main gills of S. japonicus and S. australasicus in many regions of the Atlantic and Indo-Pacific. Other species of gill monogeneans on Scomber spp. are Kuhnia scombri, Pseudokuhnia minor, Grubea cochlear and G. australis. A damaged juvenile monogenean of unknown status was recovered only once in the Mediterranean. The geographical distribution of gill monogeneans of Scomber spp. is discussed.     

Schizogony and Gametogony of Eimeria tenella in the Liver of Chick Embryos

SYNOPSIS. Schizogony and gametogony of E. tenella occurred in the liver of chick embryos inoculated intravenously with sporozoites. Gametogony occurred more freely in dexamethasonetreated embryos. Schizonts and gamonts developed within bileduct epithelial cells but many gamonts were also found within cells that appeared to be liver parenchyma type. Schizonts also developed in the chorioallantois of a corticosteroid-treated goose embryo when sporozoites were inoculated via the allantoic cavity.     

Infection of Onion by the White Rot Pathogen, Sclerotium cepivorum

Infection of onion tissue by Sclerotium cepivorum occurred from germ tubes penetrating between adjacent epidermal cell walls or directly, via penetration pegs produced from slightly swollen hyphal tips or from beneath dome shaped infection cushions. After passing through the cuticle, the infection peg enlarged to form an infection hypha within the primary cell wall. Extensive degradation of the epidermal cell wall occurred, often at a distance of 2–3 cells from the advancing hyphae. As infection advanced, hyphae spread rapidly from the epidermis to the cortex growing between and within dead/dying host cells. Extensive host cell death resulted in localized collapse of the tissue around infection points. Complete colonization of the internal tissues of the root and stem base occurred within 5–7 days of inoculation.     

Healing of skin wounds in the African catfish Clarias gariepinus

The African catfish Clarias gariepinus was used as a model for wound healing and tissue regeneration in a scale-less fish. A temporal framework of histological and cell proliferation markers was established after wound induction in the dorsolateral cranial region, by removing the epidermal and dermal layers, including stratum adiposum (SA). Wound closure and epidermis formation was initiated within 3 h post-procedure (hpp) with migration and concomitant proliferation of epidermal cells from the wound borders. The wound was covered by this primary epidermal front 12 hpp and fusion of the opposing epidermal fronts occurred within 24 hpp. Attachment of the newly formed epidermal layer to the underlying dermis was observed 48 hpp concomitant with a second wave of cell proliferation at the wound edge. Normal epidermal thickness within the wound was achieved 72 hpp. Formation of a basement membrane occurred by 120 hpp with concomitant emergence of the SA from the wound borders. Wound healing in C. gariepinus skin involved closure of the wound and re-epithelization through cell migration with a single wave of early cell proliferation not documented in other species. Furthermore, covering of the wound by epithelium as well as the reappearance of the basement membrane and SA occurred sooner than in other fish species.     

Genetic and physical characterization of recombinant plasmids associated with cell aggregation and high-frequency conjugal transfer in Streptococcus lactis ML3.

Restriction mapping was employed to characterize the 104-kilobase (kb) cointegrate lactose plasmids from 15 independent transconjugants derived from Streptococcus lactis ML3 as well as the 55-kb lactose plasmid ( pSK08 ) and a previously uncharacterized 48.4-kb plasmid ( pRS01 ) from S. lactis ML3. The data revealed that the 104-kb plasmids were cointegrates of pSK08 and pRS01 and were structurally distinct. The replicon fusion event occurred within adjacent 13.8- or 7.3-kb PvuII fragments of pSK08 and interrupted apparently random regions of pRS01 . Correlation of the transconjugants' clumping and conjugal transfer capabilities with the interrupted region of pRS01 identified pRS01 regions coding for these properties. In the 104-kb plasmids, the pRS01 region was present in both orientations with respect to the pSK08 region. The replicon fusion occurred in recombination-deficient (Rec-) strains and appeared to introduce a 0.8 to 1.0-kb segment of DNA within the junction fragments. The degeneration of the cointegrate plasmids was monitored by examining the lactose plasmids from nonclumping derivatives of clumping transconjugants. These plasmids displayed either precise or imprecise excision of pRS01 sequences or had dramatically reduced copy numbers. Both alterations occurred by rec-independent mechanisms. Alterations of a transconjugant 's clumping phenotype also occurred by rec-independent inversion of a 4.3-kb KpnI-PvuII fragment within the pRS01 sequences of the cointegrate plasmid.     

Fine structure of in situ microbial iron deposits

The fine structure of iron deposits produced by Leptothrix spp. from two caverns and a surface spring were observed. In the case of Leptothrix pseudovacuolata, the iron occurred as an amorphous deposit within the cell sheath and the cells contained gas vesicles. In the case of the other Leptothrix spp., the iron was deposited within the cell sheath either as a hexagonal or fibrillar matrix. In the case of fibrillar iron deposits, fibrils resembling those of the sheath were found within the cytoplasm of the cells. This suggests that the deposition of iron could have occurred within the cells as well as at the cell surface. In some cases, the fine structure of microbial iron deposits could provide a means of distinguishing biological from abiological iron deposition.     

Studies on encystment of Echinostoma revolutum cercariae.

Cercariae of Echinostoma revolutum encysted in the kidney of the snail Physa heterostropha within 1 hr and on mucus trails from Helisoma trivolvis, P. heterostropha and Lymnaea sp. within 2 hr. Significantly, more normal cysts were formed in mucus of Helisoma than in mucus of Physa or Lymnaea. Optimal, in vitro encystment occurred within 24 h in either Locke's 1 : 1 or Locke's 1 : 1 + 1% glucose. Significantly more normal cysts occurred in the Locke's 1 : 1 medium. Both normal and abnormal cysts from Lock's media and snail mucus excysted in an alkaline bile trypsin medium. Cercariae did not encyst in Lock'e media supplemented with casein hydrolysate or in agar cultures containing various chemicals.     

Trypanosoma (Schizotrypanum) dionisii: effect of various agents on attachment and entry to macrophages in vitro and on morphogenesis

The attachment and entry of Trypanosoma dionisii to mouse peritoneal macrophages in vitro were studied. Both occurred to a similar extent whether parasites were alive or heat-killed, and whether macrophages were obtained from normal or immunized mice. Attachment occurred equally at 4 and 37 degrees C, but entry only occurred at the higher temperature. Neither was affected by pretreatment of parasites with active or inactivated complement. Entry, but not attachment, was inhibited by cytochalasin B; both were inhibited by trypsin. Immune mouse plasma (if inactivated) stimulated attachment but not entry (within 24 h). It also stimulated intracellular replication of T. dionisii by multiple fission and subsequent differentiation (probably within macrophages) to small extracellular trypomastigotes. No extracellular parasite and only scanty intracellular forms survived 120 h in cultures containing non-inactivated immune mouse plasma. It was concluded that attachment (in the absence of antibody) occurred to non-specific receptors in the macrophage membrane and was followed by phagocytosis of the parasites rather than their active penetration of the cell.     

Emergence patterns of Plecoptera in a stream in Oregon, USA

Forty-three species of Plecoptera were collected from emergence traps in an Oregon woodland stream. Adults occurred during every month of the year, but the largest numbers and most species emerged from April to June. Temporal separation of con-generic species occurred within the suborder of Filipalpia, but not in Setipalpia. Some species of Nemoura had split emergence periods. The onset of emergence for a species from year to year generally varied by less than 2 weeks. Peak emergence occurred at least 1-2 weeks after the start ofthe emergence period.     

Operator-promoter functions in the threonine operon of Escherichia coli.

The prophage curing properties of secondary-site lysogens of coliphage lambda have been studied. The site of integration in the original lysogen (L79) is within the ooerator-promoter region of the thr operon. As a result, expression of the thr enzymes is reduced, and the strain is a leaky threonine auxotroph. Heat pulse curing of strain L79 and a thr+ lysogenic revertant (L79-20) showed that heat pulse curing of both lysogens was int and xis dependent and occurred by correct excisions of the prophage. The heat pulse curing restored strain L79 to prototrophy whereas strain L79-20 synthesized the thr enzymes constitutively and at high levels. This indicates that the reversion mutation in strain L79-20 occurred outside of the prophage and within the operator-promoter region of the thr operon. In contrast, spontaneous curing of both lysogens occurred by both correct and incorrect excisions. Spontaneously cured derivatives of strain L79-20 gave rise to three classes of regulatory mutants affecting operator and promoter functions to the thr operon.     

Vacuolar sequential exocytosis of large dense-core vesicles in adrenal medulla

Individual exocytic events in intact adrenal medulla were visualized by two-photon extracellular polar-tracer imaging. Exocytosis of chromaffin vesicles often occurred in a sequential manner, involving first vesicles located at the cell periphery and then those present deeper within the cytoplasm. Sequential exocytosis occurred preferentially at regions of the plasma membrane facing the intercellular space. The compound vesicles swelled to more than five times their original volume and formed vacuolar exocytic lumens as a result of expansion of intravesicular gels and their confinement within the lumen by the fusion pore and the narrow intercellular space. Such luminal swelling greatly promoted sequential exocytosis. The SNARE protein SNAP25 rapidly migrated from the plasma membrane to the membrane of fused vesicles. These data indicate that vesicles present deeper within the cytoplasm can be fusion ready like those at the cell periphery, and that swelling of exocytic lumens promotes assembly of the fusion machinery. We suggest the existence of two molecular configurations for fusion-ready states in Ca2+ -dependent exocytosis.