Conjugal transfer of natural plasmids between Escherichia coli strains in sterile environmental water

Seven antibiotic-multiresistant Escherichia coli strains, possessing three or four plasmids, capable of transferring their resistance marker at a high frequency, were selected among a total of 300 antibiotic-resistant E. coli strains isolated from natural water—raw and treated wastewater, and brackish water (collected 1 km downstream). These strains were mated with E. coli K-12 C600 nal^r, both in sterilized natural water and LB medium at 25°C. Conjugation did occur in all the systems tested, although fewer transconjugants were recovered from raw and treated wasterwater experiments. In contrast, in brackish and seawater, the transfer frequency did not significantly decrease in spite of salt contents. In 100% of the cases, transfer of the high-molecular-weight plasmids (20 kb) was observed, but the small plasmids (2.6–7.5 kb) were only cotransferred in raw or treated wastewater and in brackish water. Moreover, genotypic variation occurred more frequently in natural water than in LB medium.     

Five new species of coccidia (Apicomplexa: Eimeriidae) from Madagascan chameleons (Sauria: Chamaeleonidae)

Coprological examination of 19 Madagascan chameleons of the genera Furcifer and Brookesia revealed the presence of five new coccidian species. Isospora brygooi n. sp. from Furcifer pardalis has spherical to subspherical oöcysts with a slightly pitted wall, 20.7 (17–24.5) × 19.3 (16–23) m and broadly ellipsoidal sporocysts, 12.2 (11.5–13) × 8.1 (8–8.5) m, with Stieda and substieda bodies. Oöcysts of Eimeria glawi n. sp. from Furcifer pardalis are cylindrical to ellipsoidal, 27.7 (26–29.5) × 18.4 (17–19) m, with ellipsoidal sporocysts, 7.3 (6.5–8) × 5.2 (5–5.5) m. E. vencesi n. sp. described from F. pardalis has spherical to subspherical oöcysts, 14.3 (13–15.5) × 13.0 (12–13) m, with small granules, one to three globular polar granules and ellipsoidal sporocysts, 7.3 (6.5–8) × 5.2 (5–5.5) m. E. worthi n. sp., described from Furcifer oustaleti has spherical oöcysts, 17.9 (17.5–19.0) × 15.0 (14.5–16.0) m without a polar granule and ellipsoidal to cylindroidal sporocysts, 8.2 (7.0–9.5) × 5.8 (5.0–6.5) m. Oöcysts of E. brookesiae n. sp. from Brookesia decaryi are cylindrical, 25.6 (23–27) × 15.0 (13–16) m with ellipsoidal sporocysts, 10.1 (9–11) × 6.9 (6–7) m. Endogenous development of E. vencesi is confined to the intestine, while that of E. glawi occurs in the gall-bladder.     

Wardium canarisi n. sp. (Cestoda: Hymenolepididae) parasite de Arenaria melanocephala (Aves: Charadrii) d'Alaska

The authors describe and illustrate Wardium canarisi n. sp. (Cestoda: Hymenolepididae), an intestinal parasite of Arenaria melanocephala (Aves: Charadrii) from Alaska, characterised by a strobila 20–40 mm long, 10 aploparaksoid hooks 19–21 m long, a short cylindrical cirrus (40 m) covered with very minute spines (0.2 m), and a short (8–18 m) and extremely narrow (1 m) copulatory vagina. These characters have no equivalent, even approximate, among the 27 species of Wardium parasitic in the Charadrii which are reviewed. The genus Debrosia Spassky, 1987 appears to be justified.     

Taxonomic study of Ascocotyle (Phagicola) longa Ransom, 1920 (Digenea: Heterophyidae) and related taxa

Species of Ascocotyle Looss, 1899 (subgenus Phagicola Faust, 1920) (Digenea: Heterophyidae) related to A. (P.) longa Ransom, 1920, i.e. those with a bipartite gonotyl and a single crown of 16 circumoral spines, were revised on the basis of examination of type and voucher specimens. The species A. (P.) arnaldoi Travassos, 1929, A. (P.) byrdi (Robinson, 1956) and A. (P.) longicollis (Kuntz & Chandler, 1956), all possessing 16 circumoral spines, are synonymised with A. (P.) longa. A. (P.) ascolonga Witenberg, 1929, the types of which were not available, is provisionally retained as a valid species despite its close similarity to A. (P.) longa (identical number of circumoral spines and morphology of the ventrogenital sac). It differs from A. (P.) longa in the presence of uterine loops posterior to the testes and intestinal caeca ending anterior to the ovary (versus the pretesticular uterus and intestinal caeca reaching to the posterior margin of the ovary in A. (P.) longa). A. (P.) inglei (Hutton & Sogandares-Bernal, 1959), found only once in a single specimen, is also tentatively considered a valid species because it differs in number of circumoral spines (19), slightly larger eggs (23–26×13–16 m) and the distribution of refractile bodies of the gonotyl arranged in four separate groups from A. (P.) longa and A. (P.) ascolonga (16 circumoral spines; eggs 15–20×–12 m; refractile bodies in two groups).     

Expression of Ly-6A/E alloantigens in thymocyte and T-lymphocyte subsets: variability related to the Ly-6 a and Ly-6 b haplotypes

We have studied the cellular basis for differential expression of the Ly-6A/E alloantigen on T cells obtained from mice of the Ly-6 ^a (10–20% Ly-6A/E ⁺) and Ly-6 ^b (50–60% Ly-6A/E ⁺) haplotypes. During T-cell ontogeny only a small fraction (< 12 %) of thymocytes expressed Ly-6A/E. By 4 weeks of age adult levels of Ly-6A/E bearing lymphocytes were seen in peripheral lymphoid tissue. Immunohistochemical studies of the thymus revealed that Ly-6A/E⁺ cells were located predominantly in the medulla with small clusters of Ly-6A/E⁺ cells throughout the cortex. Consistent with this result, phenotypic studies showed that in the adult thymus the majority of Ly-6A/E expression was on mature CD4⁺ CD8^– and CD4^– CD8⁺ cortisone-resistant and precursor CD4^– CD8^– thymocytes. However, a much higher percentage of CD4⁺ CD8^– and CD4^– CD8^– thymocytes as well as CD4⁺ CD8^– peripheral T cells expressed Ly-6A/E from Ly-6 ^b mice. Furthermore, although gamma interferon induced increased Ly-6A/E expression in certain thymocyte and T-cell subsets, this induction functioned preferentially for cells obtained from Ly-6 ^b mice. Studies using F₁ hybrid mice (Ly-6 ^a × Ly-6 ^b) indicated that the basal level of Ly-6A/E expression on these subsets appeared to be under codominant genetic control, whereas gamma interferon-induced regulation of Ly-6A/E expression appeared to be under dominant genetic control. Collectively, these results suggest that the expression of Ly-6A/E on a particular T-cell subset is established in the thymus and is a stable characteristic of each haplotype. In addition, the low levels of Ly-6A/E expression for the Ly-6 ^a haplotype appear to be partially due to the inability of the majority of resting CD4⁺ T cells to express Ly-6A/E and to the relatively poor induction of this protein by gamma interferon.     

A daily periodicity in pituitary gonadotropin in White-crowned Sparrows

Summary Values of pituitary gonadotropic hormone (GTH) were determined throughout a 24 hour period in adult and first-year male White-crowned Sparrows (Zonotrichia leucophrys gambelii), maintained on a photoperiod of 20 hours light, 4 hours darkness. Samples taken every two hours revealed four peaks (Fig. 1) in pituitary GTH (06:00, 12:00–14:00, 18:00, 22:00). During the remaining time, GTH values were at lower levels (the minima approximately 9.0 g/gland in first-year males and 11.8 g/gland in adult males). Maximal values in adult males were from 50–125% above minimal levels, and maxima in first-year males reached 50–200% above minimal levels.Supported by NIH Predoctoral Fellowships GM-33,458 (to M.H.S.) and GM-36,159 (to J.E.E.) and by NIH Research Grant NB-06187 to Donald S. Farner. The hormones used in this investigation were generously supplied by the Endocrinology Study Section of the National Institutes of Health.     

A Comparative Study of Eubothrium Salvelini and E. crassum (Cestoda: Pseudophyllidea) Parasites of Arctic Charr and Brown Trout in Alpine Lakes

We compared the morphology, morphometry, and some aspects of the biology and ecology of Eubothrium crassum and E. salvelini, two pseudophyllidean tapeworms in brown trout, Salmo trutta m. lacustris, and Arctic charr, Salvelinus alpinus, in lakes of the western part of the Alps. The taxonomic importance of the apical disc for the discrimination of both taxa was confirmed. The apical disc of E. crassum is rectangular, possessing two deep grooves above the bothriae and 2–16 additional indentations. In E. salvelini, the apical disc is bilobed and bilaterally symmetric. We found new and significant interspecific differences in the length and width of the scolex, the diameter of the apical disc, and the width of the neck, with all measurements being larger in E. crassum. The two species differ in the number and size of the testes (fewer and larger in E. salvelini), the length of the cirrus sac (longer in E. crassum), and the size of vitelline follicles (larger in E. salvelini). Vitelline follicles of E. crassum are always cortical and sometimes enter into the outermost muscle fibres, whereas follicles of E. salvelini are situated largely medullary with few follicles entering between the innermost bundles of muscles. The eggs and the oncospheres of E. salvelini are larger. The embryonic hooks of E. crassum measure 14–18 m whilst those of E. salvelini are 18–26 m. For both taxa, the intraspecific variability in morphometric characters was fairly low. The prevalence and intensity of infection of E. crassum and E. salvelini in their respective fish hosts were very high (prevalence 90–94% mean intensity 36.3 and 6.6, maximums of 172 and 63 tapeworms per fish, respectively). E. salvelini eggs were spontaneously released throughout the year except for winter months; E. crassum laid eggs only in summer (June–August). Natural infection of copepods with Eubothrium procercoids was very low (prevalence 0.002%). The morphogenesis of Eubothrium procercoids was studied in an intermediate host, the copepod Cyclops prealpinus, that had been simultaneously infected with both species.     

Effects of varying sex pheromone component ratios on the zigzagging flight movements of the oriental fruit moth,Grapholita molesta

As the ratio of (E)-8-dodecenyl acetate (E8–12Ac) to (Z)-8-dodecenyl acetate (Z8–12Ac) increased past optimal low levels in the pheromone blend, fewer males were able to fly 2.5 m upwind to the source. The tracks of males that flew in plumes of such high-(E)off-blends were slower and narrower than those of males flying to lower-(E)blends. The tracks were narrower, first of all, because as the proportion of E8–12Ac increased, the males steered more into the wind. More of their thrust was directed upwind and therefore their groundspeed to either side of the windline was reduced. In addition, males also reduced their airspeeds to high-(E)blends, which contributed to the decreased groundspeeds and narrower tracks. No significant changes in the frequency of counterturning were found in response to increasing proportions of E8–12Ac. The inability to continue upwind flight in a plume of an off ratio was indicated by in-flight arrestment in the plume. Arrestment resulted from changes in the course angles steered by the males and the airspeeds flown.     

A Eucheuma (Solieriaceae,rhodophyta) cultivation test on the south-west coast of Madagascar

Well established populations of Eucheuma denticulatum and E. striatum exist on the coral reef of the south west coast of Madagascar but natural beds are not dense enough to support high harvest pressure. A cultivation test was conducted on a 250 m² module using the monoline method with 1560 seedlings of both species for over one year (weighings at 15 day intervals). E. denticulatum and E. striatum showed similar maximum growth rates (2.8 % d^–1) occurring respectively in fall and in summer. For both species, minimum growth was observed during the warmest months (February–March). Mean growth rates, although relatively low, showed a high standard deviation, suggesting wide possibilities of strain selection. The preliminary selection experiment led to strains more resistant to breakage and having a growth rate up to 3.3 % d^–1. The ice-ice disease is also present in this area and affects the algae mainly from January to March. Mean carrageenan yield for E. denticulatum was 42.5% ± 2.3 and rheological properties were above industrial standards, with a highly elastic gel and were thus suitable for commercial applications.     

Polymorphisms of the apolipoprotein B and E genes and their relationship to plasma lipid variables in healthy Chinese men

In this study we have analysed the apolipoprotein (Apo) E polymorphism and polymorphisms of the ApoB gene, including the ApoB/Xba I and ApoB/4311 diallelic polymorphisms and a hypervariable region (HVR) situated in the 3 region of the gene (ApoB/3HVR), in a sample of healthy male subjects from Taiyuan (northern People's Republic of China). In comparison to Caucasian populations, in the Chinese sample, the Xba I2 allele (presence of cutting site; frequency 6.1%; and 95% confidence interval, 3.3–8.9) and the long HVR alleles (9.4%; 6.0–12.8) were rare, whereas the ApoB/4311 (Ser) allele (70.8%; 65.4–76.2) and the 34-repeat allele of the HVR (HVR34; 62.4%; 56.8–68.0) were frequent. In subjects having none, one, or two HVR34 alleles, the mean levels of plasma triglycerides were 2.32±1.44 (SD), 1.45+0.74, and 1.75±1.07 g/l, respectively (P < 0.007). Similar trends were observed for very low density lipoprotein (VLDL) cholesterol, LpE:B, and LpCIII:B. The frequencies of the ApoE alleles were similar to those reported in other populations of Asian origin; E2 (7.4%; 4.2–10.6), E3 (84.4%; 80.2–88.6), and E4 (8.2%; 5.0–11.4). Individuals carrying the E2 allele had a lower mean level of ApoB than E33 individuals: 0.87±0.16 and 1.00±0.22 g/l, respectively (P < 0.007). Individuals carrying the E4 allele had higher levels of ApoE than E33 individuals: 0.140±0.084 and 0.094±0.052 g/l, respectively (P < 0.004); similar trends were observed for VLDL cholesterol, triglycerides, LpE:B, and LpCIII:B. The ApoB/ HVR34 and ApoE/E4 polymorphisms accounted for 10% to 15% of the variability of the plasma levels of VLDL cholesterol, ApoE, triglycerides, LpE:B, and LpCIII:B. Several lipid variables appeared to be favourably affected by specific forms of ApoB and ApoE that are particularly frequent in this Chinese population.     

Decolorization kinetics of a recombinant Escherichia coli strain harboring azo-dye-decolorizing determinants from Rhodococcus sp.

A 6.3 kb DNA fragment containing genes responsible for azo-dye decolorization was cloned and expressed in Escherichia coli. The resulting recombinant strain E. coli CY1 decolorized 200 mg azo dye (C.I. Reactive Red 22) l^–1 at 28 °C at 8.2 mg g cell^–1 h^–1, while the host (E. coli DH5) had no color-removal activity. Addition of 0.5 mM isopropyl--d-thiogalacto-pyranoside (IPTG) increased the decolorization rate 3.4-fold. The dependence of the decolorization rate on initial dye concentration essentially followed Monod-type kinetics and the maximal rate occurred with the dye at 600 mg l^–1. The decolorization rate of E. coli CY1 was optimal at 40 °C and pH 11. Aeration (increased dissolved O₂ level) strongly inhibited the decolorization, but decolorization occurred effectively under static incubation conditions (no agitation was employed). The CY1 strain also exhibited excellent stability during repeated-batch operations.     

Effects of selected insecticides on Cotesia plutellae, endoparasitoid of Plutella xylostella

Effects of field dosages ofselected insecticides to Cotesiaplutellae (Kurdjumov) (Hymenoptera: Braconidae), a larval endoparasitoidof Plutella xylostella L. (Lepidoptera: Plutellidae), wereinvestigated under laboratory conditions.Emergence of adult C. plutellae frominsecticide-treated pupae was not significantlydifferent from the control treatment. Contacttoxicity to C. plutellae adults variedgreatly among the insecticides in a paperresidue contact bioassay. Threeazadirachtin-based insecticides, Agroneem(4.8 mg a.i.liter^–1), Neemix (20 mga.i.liter^–1) and Ecozin (20 mgai.liter^–1) caused 11.1, 16.7 and 5.6%adult mortality, respectively. Of fourcommercial Bacillus thuringiensis (Bt)insecticides examined (all at 1.2 mga.i.liter^–1), Crymax and Xentari had noeffect on adult parasitoids, whereas Mattchcaused 5.6% mortality, and Dipel caused 11.1%mortality. Indoxacarb (53 mg a.i.liter^–1),-cyhalothrin (28 mg a.i.liter^–1) andspinosad (53 mg a.i.liter^–1) caused 100,88.5 and 50% adult mortalities, respectively.Low adult mortality (0–5.6%) was recorded fromingestion of azadirachtin-based, Btinsecticides and indoxacarb, compared with100% adult mortality in treatments of spinosador -cyhalothrin. Compared with the watercontrol, ingestion of azadirachtin-basedinsecticides significantly reduced parasitismby 50–57%, and Bt insecticides by 8–25%.However, ingestion of these insecticides didnot affect longevity of male and femaleparasitoid adults with one exception; femalelongevity was significantly reduced in theindoxacarb treatment. Insecticide residuescaused considerable mortality of C.plutellae adults, 39 and 44% mortality causedby 10 d old indoxacarb and -cyhalothrin,respectively, and 24 and 0% mortality causedby 7 and 10 d old residues of spinosad,respectively.     

Rearrangement of light harvesting bacteriochlorophyll homologues as a response of green sulfur bacteria to low light intensities

The pigment composition of two species of green-colored BChl c-containing green sulfur bacteria (Chlorobium limicola and C. chlorovibrioides) and two species of brown-colored BChl e-containing ones (C. phaeobacteroides and C. phaeovibrioides) incubated at different light intensities have been studied. All species responded to the reduction of light intensity from 50 to 1 Einstein(E) m^–2 s^–1 by an increase in the specific content of light harvesting pigments, bacteriochlorophylls and carotenoids. At critical light intensities (0.5 to 0.1 E m^–2 s^–1) only brown-colored chlorobia were able to grow, though at low specific rates (0.002 days^–1 mg prot^–1). High variations in the relative content of farnesyl-bacteriochlorophyll homologues were found, in particular BChl e ₁ and BChl e ₄, which were tentatively identified as [M, E] and [I, E] BChl_F e, respectively. The former was almost completely lost upon reduction of light intensity from 50 to 0.1 E m^–2 s^–1, whereas the latter increased from 7.2 to 38.4% and from 13.6 to 42.0% in C. phaeobacteroides and C. phaeovibrioides, respectively. This increase in the content of highly alkylated pigment molecules inside the chlorosomes of brown species is interpreted as a physiological mechanism to improve the efficiency of energy transfer towards the reaction center. This study provides some clues for understanding the physiological basis of the adaptation of brown species to extremely low light intensities.Abbreviations BChl bacteriochlorophyll - [M, E] BChl_F e 8-methyl, 12-ethyl BChl e, esterified with farnesol (F). Analogously: I - isobutyl - Pr propyl - Car carotenoids - Chlb chlorobactene - HPLC high performance liquid chromatography - Isr isorenieratene - LHP light harvesting pigments - PDA photodiode array detector - RC reaction center - RCH relative content of homologues     

Physiological responses to water stress in<Emphasis Type="Italic"> Eucalyptus cloeziana</Emphasis> and<Emphasis Type="Italic"> E. argophloia</Emphasis> seedlings

Effects of water stress duration and intensity on gas exchange and leaf water potential were investigated in 7-month-old seedlings of a humid coastal provenance (Gympie) and a dry inland (Hungry Hills) provenance of E. cloeziana F. Muell. and in a dry inland (Chinchilla) provenance of E. argophloia Blakely supplied with 100% (T₁₀₀), 70% (T₇₀), 50% (T₅₀) of their water requirements, or were watered only after they were wilted at dawn (T₀). Seedlings of E. argophloia had the highest midday net photosynthetic rate (A), stomatal conductance (g _s), stomatal density and predawn leaf water potential (_pd) in all treatments. The E. cloeziana provenances did not differ in these attributes. The T₇₀ and T₅₀ treatments caused reductions in A of 30% in E. argophloia, and 55% in the E. cloeziana provenances. Under the T₀ treatment, E. argophloia maintained higher rates of gas exchange at all levels of water stress than E. cloeziana provenances. The estimates of _pd and midday water potential (_md) at which plants remained wilted overnight were respectively: –2.7 and –4.1 MPa for E. cloeziana (humid), –2.8 and –4.0 MPa for E. cloeziana (dry) and, –3.7 and –4.9 MPa for E. argophloia. Following stress relief, both A and g _s recovered more quickly in E. argophloia and in the dry provenance of E. cloeziana than in the humid provenance. We conclude that E. argophloia is more drought tolerant and has a potential for cultivation in the humid and semi humid climates, whilst E. cloeziana has greater potential in the humid subtropical climates.     

Optimization of pyrite bioleaching using Sulfolobus acidocaldarius

Optimization of batch pyrite bioleaching with Sulfolobus acidocaldarius was performed using statistical modelling and experimental design. First a screening design was made followed by response surface modelling. The dominating factors identified were pH, pulp density and particle size. The highest batch leaching rate after optimization was 270 mg iron·l^–1·h^–1 for 6% (w/v) pulp density, pH = 1.5 and particle size <20 m. This represents a 3.5-fold increase from the leaching rate of 80 mg iron·l^–1·h^–1 obtained under our standard laboratory conditions. Correspondence to: E. B. Lindström     

Levels of toxic and essential elements in arctic fox in Svalbard

Concentrations of cadmium, mercury, lead, arsenic, selenium, copper, zinc, manganese and iron in liver, and cadmium in kidneys, were analysed in 95 carcasses of arctic fox (Alopex lagopus) caught in Svalbard during three winter seasons from 1984 through 1986. The hepatic concentration ranges of cadmium, mercury, lead and arsenic were 0.1–2.4, 0.01–2.2, < 0.5–2.9 and 0.01–1.3 g·g^–1 WW, respectively. The range of cadmium concentration in the kidneys was from 0.2 to 13 g·g^–1 WW. Cadmium and mercury concentrations were higher in adult animals than in juveniles. The average concentrations of cadmium and lead were similar to recently published levels in polar bear from Svalbard, but the mercury concentrations were lower. Significant geographical differences were observed between trapping areas. Foxes caught north of Isfjorden had lower levels of liver iron and higher levels of all other elements analysed than those caught south of Isfjorden. The recorded concentrations of heavy metals indicate a moderate degree of exposure, which most likely is of natural origin.Gunnar Norheim died January 9, 1991     

Loss of purinergic P2X receptor innervation in human detrusor and subepithelium from adults with sensory urgency

Purinergic P2X receptors associated with the parasympathetic nerves supplying human bladder smooth muscle (detrusor) are implicated in control of detrusor contractility. The relative abundance of all seven subtypes colocalised with synaptic vesicles on parasympathetic nerves was examined in specimens from normal adult bladder and in adults with the urodynamics findings of sensory urgency (SU) to determine how receptor distribution varied in patients with a small bladder capacity. Alteration in control of detrusor innervation was examined with P2X subtype-specific antibodies and an antibody (SV2) against synaptic vesicles, using immunofluorescence and confocal microscopy. Detrusor samples were taken from: controls, at cystectomy for cancer or cystoscopic biopsy for haematuria (n=22, age 33–88 years) and adults with sensory urgency at cystoscopy/cystodistension (n=11, age 37–70 years). Normal adult specimens contained detrusor muscle innervated by parasympathetic nerves possessing large varicosities (1.2 m) distributed along their length. These mostly all showed colocalised patches of presynaptic P2X_1,2,3,5 subtypes while presynaptic subtypes P2X_4,6,7 were present in only 6–18% of varicosities. Detrusor nerve varicosities from SU patients revealed general loss of all presynaptic P2X subtypes with the proportion containing receptors reducing to only 0.5–5% depending on P2X subtype. The same loss was recorded from the sensory nerves in the surrounding lamina propria. This specific loss of P2X receptors may impair control of detrusor distension and contribute to the pathophysiology of sensory urgency.The study was funded by the National Health and Medical Research Council of Australia     

Insect feeding on spores of a bracket fungus, Elfvingia applanata (Pers.) Karst. (Ganodermataceae, Aphyllophorales)

Insects visiting sporocarps of Elfvingia applanata, a wood-rotting bracket fungus, were examined in Kyoto, central Japan. Mycodrosophila flies (Drosophilidae: Diptera) were predominant and visited the spore-producing sporocarps exclusively. They were observed feeding on the spores, and a number of spores seemed to be alive even after having passed through insects digestive tracts. In addition, the insects attached a number of spores on their body surfaces. In a rearing experiment with insects caught from E. applanata sporocarps, Mycodrosophila flies excreted 7700–469 000 and dropped 10–000–329 000 of viable spores during 48 h after collection. They were supposed to migrate among the sporocarps of other bracket fungi growing on different logs or stumps, suggesting that Mycodrosophila flies may act as spore-dispersal agents for E. applanata.