Susceptibility of C5-deficient mice to listeriosis: modulation by Concanavalin A.

The susceptibility of normal (C5n) and C5 deficient (C5d) mice to Listeria monocytogenes infection and the effects of concanavalin A (Con A) on host resistance to Listeria were investigated. C5d mice were shown to be more susceptible to Listeria, but their morbidity was enhanced only with certain doses, 2 to 6 × 10⁴ viable Listeria. Listeria infection produced immune cells in both C5n and C5d mice as assessed in a passive transfer study, but the C5d immune cells significantly suppressed Listeria growth only when transferred into C5n mice. Con A altered the response to Listeria in both C5n and C5d mice. The growth of Listeria in the spleens of C5n and C5d mice, which were treated with Con A 1 day before infection, was clearly suppressed, and their mortality was reduced. Con A given 1 day after infection (Day +1) consistently enhanced the early (Days 1 to 3 post-infection) accumulation of Listeria in the spleens, and A/J (C5d) and CBA/J (C5n) mortality was increased but B10.D2 (C5d and C5n) mortality was reduced. Con A given 7 days before Listeria infection produced no differential effects on the early growth of Listeria in the spleens of C5n and C5d mice, but most C5n mice survived a lethal dose of Listeria and the C5d mice died. Since C5n and C5d mice differ in their resistance to Listeria, Con A (Day ?7), a selective T-cell mitogen, produced differential effects in C5n and C5d mice and Listeria is believed to induce predominantly a cell-mediated imune response, T cells and complement components independently or cooperatively appear to play a role in immunity to Listeria.     

Duration of development and longevity inAphidius ervi andAphidius platensis [Hym.: Aphidiidae], two parasites ofMyzus persicae [Hom.: Aphididae]

Rate of development and longevity were studied inAphidius ervi Haliday andAphidius platensis Brèthes, two parasites of the green peach aphid,Myzus persicae Sulz. On paprika,A. ervi developed from egg to adult in 27.3 days (at 15°C) and 19.9 days (21°C),A. platensis in 19.9 days (15°C), 15.6 days (21°C) and 12.4 days (24°C). The period from oviposition to mummification was in both species roughly twice as long as the period from mummification to adult emergence. Males emerged slightly before females. Given water and honey,A. ervi lived 15.4 days (♂) and 13.1 days (♀) at 21°C. The effect of temperature on longevity was tested inA. platensis on this diet: at 15°C, 9.2 days (♂) and 7.9 days (♀); at 21°C, 12.0 days (♂) and 13.4 days (♀); and at 24°C, 7.4 days (♂) and 8.4 days (♀). When supplied only with water, both species lived for 1–3 days. When aphid-infested leaves were added, longevity increased by 3.5 days (A. platensis). The maximal longevity, obtained with water and honey, was somewhat reduced when leaves were added, probably due to mating and oviposition activities (A. ervi). Longevity was not significantly influenced by the different host plants during parasite development. Differences in longevity between the sexes were small and dependent on temperature and food.     

The systematics of clarkia bottae,C. Cylindrica,and a related new species,C. Rostrata

Populations ofClarkia cylindrica andC. bottae were studied in the field and in the garden. Seed from which the garden progenies were grown was collected in the wild. Experimental crosses were made between and within taxa. Fertility and chromosome pairing were observed in the hybrids. A maternal influence on seed set was found in crosses between C.cylindrica andC. bottae. Two taxa are described as new:Clarkia rostrata andC. cylindrica ssp.clavicarpa.     

Phylogenetic analyses of Japanese golden chanterelles and a new species description,Cantharellus anzutake sp. nov.

The Japanese golden chanterelle commonly identified as Cantharellus cibarius was sampled in a broad range of forest vegetation. A total of 90 fresh and 11 herbarium specimens were examined microscopically, subjected to sequencing analysis of their nuclear ribosomal RNA (rDNA) and tef-1 genes, and their characteristics were compared with those of European C. cibarius. Based on morphological and ecological characteristics, basidioma samples from Japan were divided into four species. While specimens of Cantharellus sp. 4 from Hokkaido Island were included in the European C. cibarius clade phylogenetically, the other three species formed three unique clades. Among these, Cantharellus anzutake sp. nov. is sister to the clade of C. cibarius and was widely sampled from the northern limit of Honshu Island to the southern limit of Kumejima Island in Ryukyu Islands. Although C. anzutake was morphologically similar to C. cibarius, the two species were phylogenetically distinct. Other morphologically similar but genetically distinct chanterelle species from India exhibited macroscopic and microscopic differences compared with C. anzutake.     

Cell surface differentiation of Chlamydomonas during gametogenesis. I. Mating and concanavalin A agglutinability

Sexual agglutination of opposite gametes and agglutination of like gametes by concanavalin A (Con A) were studied in Chlamydomonas moewusii and C. reinhardtii for the possibility that the surface sites involved in these agglutinating phenomena may be the same. Our data show that the two agglutinating phenomena appeared at different times after the beginning of gametogenesis. Sucrose and mannose block agglutination of the gametes by Con A but do not affect mating ability. Trypsin eliminates mating ability, except in (?) gametes of C. moewusii, while Con A agglutinability remains. Monovalent Con A can selectively bind to Con A-binding sites to block agglutination of gametes by multivalent Con A while mating ability is unaffected. The data indicate that the mating agglutinin and the Con A-binding sites are two different flagellar surface agglutinins that occur coincidentally during gametic differentiation of both mating types of C. moewusii and C. reinhardtii. The function of the Con A-binding site on Chlamydomonas gametes is not known.     

Comparison of the Thermostability Properties of Three Acid Phosphatases from Molds: Aspergillus fumigatus Phytase, A. niger Phytase, and A. niger pH 2.5 Acid Phosphatase

Enzymes that are used as animal feed supplements should be able to withstand temperatures of 60 to 90°C, which may be reached during the feed pelleting process. The thermostability properties of three histidine acid phosphatases, Aspergillus fumigatus phytase, Aspergillus niger phytase, and A. niger optimum pH 2.5 acid phosphatase, were investigated by measuring circular dichroism, fluorescence, and enzymatic activity. The phytases of A. fumigatus and A. niger were both denatured at temperatures between 50 and 70°C. After heat denaturation at temperatures up to 90°C, A. fumigatus phytase refolded completely into a nativelike, fully active conformation, while in the case of A. niger phytase exposure to 55 to 90°C was associated with an irreversible conformational change and with losses in enzymatic activity of 70 to 80%. In contrast to these two phytases, A. niger pH 2.5 acid phosphatase displayed considerably higher thermostability; denaturation, conformational changes, and irreversible inactivation were observed only at temperatures of ≥80°C. In feed pelleting experiments performed at 75°C, the recoveries of the enzymatic activities of the three acid phosphatases were similar (63 to 73%). At 85°C, however, the recovery of enzymatic activity was considerably higher for A. fumigatus phytase (51%) than for A. niger phytase (31%) or pH 2.5 acid phosphatase (14%). These findings confirm that A. niger pH 2.5 acid phosphatase is irreversibly inactivated at temperatures above 80°C and that the capacity of A. fumigatus phytase to refold properly after heat denaturation may favorably affect its pelleting stability.     

A new wilt and die-back disease of Acacia mangium associated with Ceratocystis manginecans and C. acaciivora sp. nov. in Indonesia

Species of Ceratocystis are well-known wound related pathogens of many tree species, including commercially planted Acacia spp. Recently, several Ceratocystis isolates were collected from wilting A. mangium in plantations in Indonesia. The aim of this study was to identify these Ceratocystis isolates and to investigate their ability to cause disease on two plantation-grown Acacia spp. using greenhouse and field inoculation experiments. For identification, morphological characteristics and comparisons of DNA sequence data for the ITS, β-tubulin and TEF 1-α gene regions, was used. Ceratocystis isolates were identified as C. manginecans, a serious pathogen of mango trees in Oman and Pakistan and a previously undescribed species, described here as C. acaciivora sp. nov. Both fungi produced significant lesions in inoculation experiments on A. mangium and A. crassicarpa, however, C. acaciivora was most pathogenic suggesting that this fungus is the primary cause of the death of trees under natural conditions.     

Cryptosporidium apodemi sp. n. and Cryptosporidium ditrichi sp. n. (Apicomplexa: Cryptosporidiidae) in Apodemus spp.

Faecal samples from striped field mice (n = 72) and yellow-necked mice (n = 246) were screened for Cryptosporidium by microscopy and PCR/sequencing. Phylogenetic analysis of small-subunit rRNA, Cryptosporidium oocyst wall protein and actin gene sequences revealed the presence of C. parvum, C. hominis, C. muris and two new species, C. apodemi and C. ditrichi. Oocysts of C. apodemi are smaller than C. ditrichi and both are experimentally infectious for yellow-necked mice but not for common voles. Additionally, infection by C. ditrichi was established in one of three BALB/c mice. The prepatent period was 7–9 and 5–6 days post infection for C. apodemi and C. ditrichi, respectively. The patent period was greater than 30 days for both species. Infection intensity of C. ditrichi ranged from 4000–50,000 oocyst per gram of faeces and developmental stages of C. ditrichi were detected in the jejunum and ileum. In contrast, neither oocysts nor endogenous developmental stages of C. apodemi were detected in faecal or tissue samples, although C. apodemi DNA was detected in contents from the small and large intestine. Morphological, genetic, and biological data support the establishment of C. apodemi and C. ditrichi as a separate species of the genus Cryptosporidium.     

Genetic Diversity of Cryptosporidium spp. in Captive Reptiles

The genetic diversity of Cryptosporidium in reptiles was analyzed by PCR-restriction fragment length polymorphism and sequence analysis of the small subunit rRNA gene. A total of 123 samples were analyzed, of which 48 snake samples, 24 lizard samples, and 3 tortoise samples were positive for Cryptosporidium. Nine different types of Cryptosporidium were found, including Cryptosporidium serpentis, Cryptosporidium desert monitor genotype, Cryptosporidium muris, Cryptosporidium parvum bovine and mouse genotypes, one C. serpentis-like parasite in a lizard, two new Cryptosporidium spp. in snakes, and one new Cryptosporidium sp. in tortoises. C. serpentis and the desert monitor genotype were the most common parasites and were found in both snakes and lizards, whereas the C. muris and C. parvum parasites detected were probably the result of ingestion of infected rodents. Sequence and biologic characterizations indicated that the desert monitor genotype was Cryptosporidium saurophilum. Two host-adapted C. serpentis genotypes were found in snakes and lizards.     

The cryopreservation of Chlamydomonas.

A cryophilic strain of the unicellular green alga Chlamydomonas, C. nivalis was found to be more resistant to the stresses both of freezing and thawing and of shrinkage and rehydration than was a mesophilic strain C. reinhardii. C. nivalis was found to have a higher degree of unsaturation of phospholipid fatty acids. Following freezing and thawing of C. reinhardii there was a direct correlation between reduction in cell viability and loss of membrane selective permeability. Activation of intracellular phospholipases occurred at an early stage of freezing injury. Attempts to cold harden C. reinhardii were unsuccessful. For C. reinhardii methanol was the only effective cryoprotectant for freezing to and thawing from ?196 °C and the effects of cooling rate upon cellular survival are presented.     

Analysis of morphological characters of Chara baltica, C. hispida, C. horrida, and C. rudis from Europe

Morphological characters of Chara baltica, C. hispida, C. horrida, and C. rudis from different localities in Central Europe and Scandinavia were analyzed. The plants were preliminarily classified using specific qualitative features into four species: C. baltica, C. hispida, C. horrida, and C. rudis. Of the 14 characters, all were used in principal-component and cluster analyses, and 12 were applied in a discriminant analysis. In the discriminant analysis, the most useful characters for distinguishing particular species were selected. The methods (canonical discriminant and canonical analyses) used herein offered good differentiation between brackish water species C. horrida and C. baltica and fresh water species C. hispida and C. rudis. The analysis showed that the two morphologically very similar species C. hispida and C. rudis are only partially distinguished.     

Genetic studies of the lac repressor. IV. Mutagenic specificity in the lacI gene of Escherichia coli.

An extensive set of amber and ochre sites in the lacI gene has been characterized with respect to the base change required to generate the nonsense codon (Miller et al., 1977; Coulondre &; Miller, 1977). These mutations have been used to analyze the forward mutational spectrum of a series of mutagens in Escherichia coli. The sites induced by N′-methyl-N′-nitro-N-nitrosoguanidine, ethyl methanesulfonate, 4-nitroquinoline-1-oxide, and ultraviolet light, were examined, as well as those which arose spontaneously. Sites induced by the G · C → A · T transition were compared with those generated by 2-aminopurine mutagenesis. All together, more than 4000 independent occurrences of amber and ochre mutations were tabulated in order to define the respective mutagenic specificities. With the exception of the A · T → G · C change, all base substitutions lead to the generation of nonsense codons from wild-type. The A · T → G · C transition was monitored in a reversion system, in which the ochre to amber conversion (UAA → UAG) was scored, as well as the UAA → CAA reversion.Both NG³ and EMS were found to be highly specific for the G · C → A · T transition, less than 1% transversions appearing in either case. At between 1% and 5% the level of the G · C → A · T change, NG can stimulate the A · T → G · C transition. EMS stimulates the A · T → G · C transition at a significantly lower rate. NQO is also highly specific for G · C base-pairs, but approximately 10% of the changes found at these sites are transversions. Mutations found spontaneously or after irradiation with ultraviolet light showed none of the specificities found for EMS, NG or NQO. All transversions were detected in both cases. Moreover, a significant number of tandem double base changes were found to be induced by u.v. irradiation. Some of these have been verified directly by protein sequencing. The frequencies of occurrence of amber and ochre mutations arising from the G · C → A · T transition have been compared for different mutagens, revealing several striking hotspots. The implications of these findings with respect to the mechanism of mutagenesis and the application of different mutagens are discussed.     

Pseudomonas versuta sp. nov., isolated from Antarctic soil

In this study we analysed three bacterial strains coded L10.10^T, A4R1.5 and A4R1.12, isolated in the course of a study of quorum-quenching bacteria occurring in Antarctic soil. The 16S rRNA gene sequence was identical in the three strains and showed 99.7% pairwise similarity with respect to the closest related species Pseudomonas weihenstephanensis WS4993^T. Therefore, the three strains were classified within the genus Pseudomonas. Analysis of housekeeping genes (rpoB, rpoD and gyrB) sequences showed similarities of 84-95% with respect to the closest related species of Pseudomonas, confirming its phylogenetic affiliation. The ANI values were less than 86% to the closest related species type strains. The respiratory quinone is Q9. The major fatty acids are C16:0, C16:1 ω7c/ C16:1 ω6c in summed feature 3 and C18:1 ω7c / C18:1 ω6c in summed feature 8. The strains are oxidase- and catalase-positive. Growth occurs at 4–30 °C, and at pH 4.0–10. The DNA G+C content is 58.2–58.3 mol %. The combined genotypic, phenotypic and chemotaxonomic data support the classification of strains L10.10^T, A4R1.5 and A4R1.12 into a novel species of Pseudomonas, for which the name P. versuta sp. nov. is proposed. The type strain is L10.10^T (LMG 29628^T, DSM 101070^T).     

Pollen morphology of the aquatic Brazilian endemic genus Castelnavia Tul. & Wedd. (Podostemaceae)

The utility of pollen characteristics for taxonomic and phylogenetic analysis of most genera of Podostemaceae has barely been examined. Herein we report on the diversity of pollen structure in Castelnavia (Podostemaceae), a genus of six species endemic to central and southern Brazil. Pollen grain was shed as three-colpate, planaperturate monads and ranged from 12.5 to 22.5?μm for the polar axis and from 12.5 to 20.0?μm for the equatorial axis. All species examined had polar and radially symmetrical pollen grains. Grains from Castelnavia fluitans, Castelnavia multipartita, and Castelnavia noveloi had an oblate spheroidal shape (PD/ED), those from Castelnavia monandra and Castelnavia pendulosa were subprolate, and those from Castelnavia princeps were prolate spheroidal. The polar view was subcircular for C. monandra and subtriangular for the other species. A fastigium was observed in C. fluitans and C. pendulosa whereas a costa was present in C. multipartita and C. princeps, with the colpus margin not thickened in the former species but prominently thickened in the latter. Castelnavia monandra and C. noveloi did not have a fastigium or costa. For all species spicules occurred on the exine (microechinate sexine) and were most conspicuous on the colpus membrane. There was little correspondence between variation in pollen structure and species relationships supported by phylogenetic analysis.     

Molecular and Phenotypic Analyses Reveal Association of Diverse Colletotrichum acutatum Groups and a Low Level of C. gloeosporioides with Olive Anthracnose

Anthracnose (Colletotrichum spp.) is an important disease causing major yield losses and poor oil quality in olives. The objectives were to determine the diversity and distribution pattern of Colletotrichum spp. populations prevalent in olives and their relatedness to anthracnose pathogens in other hosts, assess their pathogenic variability and host preference, and develop diagnostic tools. A total of 128 Colletotrichum spp. isolates representing all olive-growing areas in Portugal and a few isolates from other countries were characterized by molecular and phenotypic assays and compared with reference isolates. Arbitrarily primed PCR data, internal transcribed spacer of rRNA gene and β-tubulin 2 nucleotide sequences, colony characteristics, and benomyl sensitivity showed Colletotrichum acutatum to be dominant (>97%) with limited occurrence of Colletotrichum gloeosporioides (<3%). Among C. acutatum populations, five molecular groups, A2 to A6, were identified. A2 was widely prevalent (89%), coinciding with a high incidence of anthracnose and environmental conditions suitable to disease spread. A4 was dominant in a particular region, while other C. acutatum groups and C. gloeosporioides were sporadic in their occurrence, mostly related to marginal areas of olive cultivation. C. gloeosporioides, isolated from olive fruits with symptoms indistinguishable from those of C. acutatum, showed same virulence rating as the most virulent C. acutatum isolate from group A2. C. acutatum and C. gloeosporioides isolates tested in infected strawberry fruits and strawberry and lupin plants revealed their cross-infection potential. Diagnostic tools were developed from β-tubulin 2 sequences to enable rapid and reliable pathogen detection and differentiation of C. acutatum groups.     

Reproductive biology of Fopius arisanus (Hymenoptera: Braconidae) on Ceratitis capitata and Anastrepha spp. (Diptera: Tephritidae)

The reproduction of the solitary endoparasitoid Fopius arisanus (Sonan) (Hymenoptera: Braconidae) in Anastrepha ludens (Loew), Anastrepha obliqua (Macquart), and Anastrepha serpentina (Wiedemann) was compared with that using Ceratitis capitata (Wiedemann), being the host in which it had been reared previously. Eggs of different ages (<4 h old, 1, 2, and 3 days old for Anastrepha spp., and ⩽4 h, 1 and 2 days old for C. capitata) of each host species were placed in pieces of papaya, exposed to parasitism for 24 h and then reared through to the adult stage. Host species had a marked effect on parasitoid reproduction with consistently higher parasitoid emergence from C. capitata, whereas emergence from A. obliqua was negligible and this host was not studied further. Host age did not significantly affect parasitoid emergence from C. capitata whereas parasitism of A. ludens and A. serpentina was significantly greater in eggs exposed at 3 days old than those exposed at younger ages. Adult parasitoid sex ratio was male biased in all cases. Despite significant differences in host developmental time, host species did not affect parasitoid developmental time. Parasitoid life expectancy at emergence was reduced by >60% for parasitoids that emerged from A. ludens compared to those that emerged from A. serpentina or C. capitata. The reproduction of parasitoid progeny was highest in parasitoids that emerged from and reproduced on C. capitata and lowest for parasitoids reproducing on A. ludens. Parasitoids that emerged from A. ludens were often deformed, but were larger than those that emerged from A. serpentina. Parasitoids that emerged from C. capitata were smaller than those from Anastrepha spp. We conclude that F. arisanus is capable of sustained reproduction in C. capitata and A. serpentina and merits further study as an agent for the control of these fruit flies.     

Aflatoxin Production in Meats. II. Aged Dry Salamis and Aged Country Cured Hams

Italian-type salamis contaminated with Aspergillus flavus were more likely to develop aflatoxins during aging than were smoked Hungarian-type salamis under the same conditions. Temperatures below 15 C and humidities of less than 75% were found to prevent aflatoxin development during the aging of salami. The aging of salami for 8 weeks and the presence of curing ingredients, especially pepper and sodium nitrite, tended to reduce the amounts of aflatoxins found. Aflatoxins were produced by A. flavus and A. parasiticus on 6- to 9-month-old country cured hams during aging when the temperature approached 30 C.     

A new putrescine bisamide phenolic glycoside from the seeds of Lallemantia iberica (M. Bieb.) Fisch. & C. A. Mey

A new putrescine bisamide phenolic glycoside, N-(trans-feruloyl)-N′-(para-hydroxybenzoyl) putrescine bisamide-4′-O-α-l-rhamnopyranoside (1), designated as lallenmantoside (1), together with one known phenolic glycoside, cucurbitoside d (2), were isolated from the seeds of Lallemantia iberica (M. Bieb.) Fisch. & C. A. Mey. Their structures were elucidated by spectroscopic analysis, and by comparison with literature data. This is the first report of phenolic glycosides from genus Lallemantia.