Seasonal variations in the immune system of the tench,Tinca tinca (Cyprinidae): proliferative response of lymphocytes induced by mitogens

The proliferation of tench lymphocytes induced by mitogens was studied during the four seasons of the year. Fish were maintained under natural conditions of photoperiod and temperature (mean ± SD: 12±2°C in winter, 22±3°C in spring, 30±3°C in summer and 21±3°C in autumn). Cultures were performed in vitro at 22°C in all seasons and the results were compared. Subsequently, in seasons with extreme water temperatures, cultures in vitro were performed at the same temperature as that of the water (12°C in winter and 30°C in summer) and the results were compared seasonally at the seasonal temperature, i.e. at 22°C in spring, 30°C in summer, 22°C in autumn and 12°C in winter. Phytohemagglutinin, concanavalin A, lipolisaccharide from E. coli and pokeweed mitogen were used as mitogens. Studies performed at 22°C as assay temperature in all seasons showed profound seasonal changes: while in spring, summer and autumn the mitogenic response of lymphocytes to phytohemagglutinin, concanavalin A, lipolisaccharide from E. coli and pokeweed mitogen was very low, during winter the results obtained were significantly higher. However, when the assays were performed at the corresponding seasonal temperature the differences were not as pronounced between the different seasons, and the mitogenic responses of lymphocytes were found to be the lowest during the winter and the highest during the summer with all mitogens used. This fact suggests that immunosuppression occurs in winter and an immunostimulation occurs in summer. However, the higher response found in winter when assaying at 22°C suggests that this property of lymphocytes needs an assay temperature higher than the in vivo temperature in order to observe accurate mitogenic responses.Abbreviations Con A concanavalin A - cpm counts per minute - LPS E. coli lipolisaccharide - MS222 tricainemethane sulphonate - PBS phosphate-buffered saline - PHA phytohemagglutinin - PWM pokeweed mitogen - SI stimulation index     

Seasonal changes in phagocytic capacity and superoxide anion production of blood phagocytes from tench (Tinca tinca,L.)

Seasonal variations in the ex vivo phagocytic function of blood cells from tench, including ingestion capacity of inert particles and its destruction (microbicide capacity) assessed by measurement of superoxide anion production, were studied. Tench were maintained under natural conditions throughout the year, and the different assays of samples taken during each season were initially performed in vitro at 22°C and the results compared. Subsequently, assays were performed at the same temperature as that of the water ponds in which the fish were kept (“seasonal temperature”: 12°C in winter, 22°C in spring and autumn and 30°C in summer) and the results compared seasonally. The results at 22°C showed that phagocytic capacity was greatest in spring and summer and lowest in winter. However, when phagocytic capacity was measured at seasonal temperature, highest values appeared in winter and lowest in summer and autumn. Nitroblue tetrazolium reduction by tench phagocytes after phagocytosing latex beads demonstrated a similar seasonal behaviour at both 22°C in each season and at seasonal temperature. The highest values appeared in summer, which suggests a better microbicide capacity in this season. The results obtained in this study suggest that for a correct interpretation of ex vivo phagocytic capacity of fish through the year it is necessary to use the same assay temperature as that of the water in which the fish is kept.     

The ORF encoding a putative ferredoxin-like protein downstream of the vnfH gene in Azotobacter vinelandii is involved in the vanadium-dependent alternative pathway of nitrogen fixation

Summary An open reading frame (ORF) in the same operon as, but downstream of, vnfH in Azotobacter vinelandii can code for a ferredoxin-like protein. The role this ORF may play in the vnf (vanadium-dependent alternative) pathway of nitrogen fixation was investigated. Site-directed mutagenesis was used to alter one base in each of the codons specifying amino acids 18 and 19 generating a unique BglII site. A kanamycin resistance cartridge was cloned into the BglII site. This construct was mobilized into A. vinelandii CA12 ( nifHDK) strain by conjugation and the mutation was introduced into the genome by marker exchange. The resulting mutant was unable to fix nitrogen under conditions in which the vnf pathway of nitrogen fixation operates. This suggests that this ORF is functional and is essential for the vanadium-dependent alternative pathway of nitrogen fixation in A. vinelandii.     

葱属植物S-烷(烯)基半胱氨酸亚砜代谢途径研究进展

葱属植物是被子植物中最大的属之一，包括大蒜、洋葱、大葱、韭菜等多种具有独特辛辣风味的蔬菜作物。S-烷(烯)基半胱氨酸亚砜是葱属植物特有的次生代谢产物，是葱属植物各种挥发性含硫化合物的前体物质，由于其赋予葱属植物独特的辛辣风味和药用价值，因此研究葱属植物S-烷(烯)基半胱氨酸亚砜的代谢途径具有重要意义。在葱属植物中已发现7种S-烷(烯)基半胱氨酸亚砜，这些S-烷(烯)基半胱氨酸亚砜主要在叶片中先经谷胱甘肽途径合成，再转运到鳞茎等贮藏器官的细胞质中积累。目前，关于葱属植物S-烷(烯)基半胱氨酸亚砜降解的研究较多，而S-烷(烯)基半胱氨酸亚砜生物合成的研究则较少。S-烷(烯)基半胱氨酸亚砜是植物硫代谢的下游产物，上游涉及含硫化合物的吸收、转运、半胱氨酸和谷胱甘肽的代谢等过程，这些代谢过程的变化可能影响S-烷(烯)基半胱氨酸亚砜生物合成。今后，应加强以下两方面的研究：一方面，继续克隆鉴定S-烷(烯)基半胱氨酸亚砜生物合成途径中的关键酶基因，并研究其功能；另一方面，加强葱属植物硫代谢的研究，为研究S-烷(烯)基半胱氨酸亚砜生物合成的调控奠定基础。该研究结果为深入解析葱属植物S-烷(烯)基半胱氨酸...     

Seasonal variations in carbon and nitrogen content of Acartia clausi (Copepoda,Calanoida) in the Gulf of Trieste (Northern Adriatic Sea)

A yearly study was made on total and cephalothorax lengths, dry weight, carbon and nitrogen contents and C:N ratio of the species Acartia clausi (Copepoda, Calanoida) which is always abundant, particularly in spring, in the net zooplankton community of the Gulf of Trieste (Northern Adriatic Sea). The samples of net zooplankton were carried out every month at the permanent station located 200 m offshore and preserved for about one year in 4% buffered formalin to insure weight loss stabilization. A. clausi was separated from other zooplankton specimens; washed and dried. For each monthly sample the average total and cephalothorax lengths and the average dry weight of adult organisms of A. clausi were measured. The mean carbon and nitrogen contents, as a percentage of dry weight, were determined by using a CHN analyser. Linear regression models were computed on the log-transformed data in order to check the relationships between dry weight, total and cephalothorax lengths, carbon and nitrogen contents and C:N ratio of A. clausi. The best of the various established correlations, was between dry weight and carbon content. Our study pinpointed a high seasonal variability of the C:N ratio, mainly due to seasonal fluctuations in the nitrogen content.     

Seasonal reversibility of acclimation to irradiance in leaves of common box (Buxus sempervirens L.) in a deciduous forest

Understorey shade plants are seasonally exposed to dramatic changes in light conditions in deciduous forests related with the dynamics of the overstorey leaf phenology. These transitions are commonly followed by changes in herb plant communities, but shade-tolerant evergreen species must be able to adapt to changing light conditions. In this work we checked the photoprotective responses of evergreen species to acclimate to the shady summer environment and reversibly de-acclimate to a more illuminated environment after leaf fall on deciduous overstoreys. For that purpose we have followed the process of light acclimation in leaves of common box (Buxus sempervirens) during the winter to spring transition, which decrease irradiance in the understorey, and conversely during the transition from summer to autumn. Four parameters indicative of the structure and degree of acclimation of the photosynthetic apparatus have been studied: chlorophyll a/b ratio which is supposed to be inversely proportional to the antenna size, α/β-carotene which increases in shade acclimated leaves and the pools of α-tocopherol and xanthophyll cycle pigments (VAZ) which are two of the main photoprotection mechanisms in plants. Among these parameters, chlorophyll a/b ratio and VAZ pool responded finely to changes in irradiance indicating that modifications in the light harvesting size and photoprotective capacity contribute to the continuous acclimation and de-acclimation of long-lived evergreen leaves.     

Seasonal changes in a population of Menacanthuseurysternus (mallophaga, amblycera) on the common myna Acridotheres tristis

Seasonal changes in a population of Menacanthus eurysternus (Mallophaga, Amblycera) on the common myna Acridotheres trislis. International Journal for Parasitology20: 1063–1065. Seasonal variations in a population of an amblyceran louse, Menacanthus eurysternus, infesting the common myna have been studied. Incidence of infestation, lice, index, and population structure of this species have been recorded in different months of the year; reasons for the population fluctuations are discussed.     

Distribution of intravenously injected A-layer protein and lipopolysaccharide (LPS) from Aeromonas salmonicida in Atlantic salmon, Salmo salar L.

The distribution of intravenously injected A-layer protein and lipopolysaccharide (LPS) purified from the outer surface of the fish pathogen Aeromonas salmonicida, was studied in Atlantic salmon. Radiolabelling was achieved by conjugating the antigens to tyramine cellobiose (TC) or fluorescein isothiocyanate (FITC) which were radioiodinated either before or after conjugation. Since both TC and FITC are trapped intralysosomally at the cellular site of uptake, the ligands are advantageous in studies on tissue distribution of antigens. Injection of TC-A-layer protein and TC-LPS resulted in high specific radioactivity (cpm g⁻¹tissue) in both head kidney and trunk kidney. In contrast, only low specific radioactivity was recovered in spleen, heart and liver. Surprisingly, use of FITC-LPS as the antigen changed the uptake to be high in both spleen and head kidney. Radiolabelled (¹²⁵I-TC-) LPS and A-protein, administered by a dorsal aorta catheterisation technique, were cleared from the blood within 24 h. In immunised fish, the antibody activity against the A-layer protein was diminished even within 10 min after administration, in contrast to the level of anti-LPS antibodies which remained high. These results suggest that immune-complex formation took place at least with the A-layer protein, but the uptake of A-layer protein in the various tissues did not differ significantly in vaccinated (A. salmonicida bacterin) and non-vaccinated fish.     

MSl3, a multicopy suppressor of mutants hyperactivated in the RAS-CAMP pathway, encodes a novel HSP70 protein of Saccharomyces cerevisiae

The MSI3 gene was isolated as a multicopy suppressor of the heat shock-sensitive phenotype of the iral mutation, which causes hyperactivation of the RAS-cAMP pathway. Overexpression of MSI3 also suppresses the heat shock-sensitive phenotype of the bcyl mutant. Determination of the DNA sequence of MSI3 revealed that MSI3 can encode a 77.4 kDa protein related to the HSP70 family. The amino acid sequence of Msi3p is about 30% identical to that of the Ssalp of Saccharomyces cerevisiae. This contrasts with the finding that members of the HSP70 family generally show at least 50% amino acid identity. The consensus nucleotide sequence of the heat shock element (HSE) was found in the upstream region of MSI3. Moreover, the steady-state levels of the MSI3 mRNA and protein were increased upon heat shock. These results indicate that the MSI3 gene encodes a novel HSP70-like heat shock protein. Disruption of the MSI3 gene was associated with a temperature sensitive growth phenotype but unexpectedly, thermotolerance was enhanced in the disruptant.     

Seasonal variations in the behavioural thermoregulation of roosting Humboldt penguins ( Spheniscus humboldti) in north-central Chile

We examined the thermoregulatory behaviour (TRB) of roosting Humboldt penguins (Spheniscus humboldti) in north central Chile during summer and winter, when ambient temperatures (T_a) are most extreme. Each body posture was considered to represent a particular TRB, which was ranked in a sequence that reflected different degrees of thermal load and was assigned an arbitrary thermoregulatory score. During summer, birds exhibited eight different TRBs, mainly oriented to heat dissipation, and experienced a wide range of T_a (from 14 to 31°C), occasionally above their thermoneutral zone (TNZ, from 2 to 30°C), this being evident by observations of extreme thermoregulatory responses such as panting. In winter, birds exhibited only three TRBs, mainly oriented to heat retention, and experienced a smaller range of T_a (from 11 to 18°C), always within the TNZ, even at night. The components of behavioural responses increased directly with the heat load which explains the broader behavioural repertoire observed in summer. Since penguins are primarily adapted in morphology and physiology to cope with low water temperatures, our results suggest that behavioural thermoregulation may be important in the maintenance of the thermal balance in Humboldt penguins while on land.     

Spatio-temporal variations of functional groups in a Populus nigra L. entomocenosis in the Mitidja plain (Algeria)

This study presents the first data describing the communities of insects and arachnida colonizing 4 aerial compartments of Populus nigra. In terms of temporal variation and total abundance, the analyses show that the communities are structurally and functionally different. It also appears that the taxonomic richness of the communities, and partly their temporal distribution, change according to the gradient of the energy and defense resources under the effect of seasonal variations. The highest species richness recorded on the leaves compartment is due to the amplitude of histo-physiological modifications observed throughout the growing season. To cite this article: Z.-E. Djazouli et al., C. R. Biologies 332 (2009).     

Molecular characterization of the alpha subunit of complement component C8 (GcC8α) in the nurse shark (Ginglymostoma cirratum)

Target cell lysis by complement is achieved by the assembly and insertion of the membrane attack complex (MAC) composed of glycoproteins C5b through C9. The lytic activity of shark complement involves functional analogues of mammalian C8 and C9. Mammalian C8 is composed of α, β, and γ subunits. The subunit structure of shark C8 is not known. This report describes a 2341 nucleotide sequence that translates into a polypeptide of 589 amino acid residues, orthologue to mammalian C8α and has the same modular architecture with conserved cysteines forming the peptide bond backbone. The C8γ-binding cysteine is conserved in the perforin-like domain. Hydrophobicity profile indicates the presence of hydrophobic residues essential for membrane insertion. It shares 41.1% and 47.4% identity with human and Xenopus C8α respectively. Southern blot analysis showed GcC8α exists as a single copy gene expressed in most tissues except the spleen with the liver being the main site of synthesis. Phylogenetic analysis places it in a clade with C8α orthologs and as a sister taxa to the Xenopus.     

Characterization of a chromosomally encoded, non-PTS metabolic pathway for sucrose utilization in Escherichia coli EC3132

Summary A wild-type isolate, EC3132, of Escherichia coli, that is able to grow on sucrose was isolated and its csc genes (mnemonic for chromosomally coded sucrose genes) transferred to strains of E. coli K12. EC3132 and all sucrose-positive exconjugants and transductants invariably showed a D-serine deaminase (Dsd)-negative phenotype. The csc locus maps adjacent to dsdA, the structural gene for the D-serine deaminase, and contains an inducible regulon, controlled by a sucrose-specific repressor CscR, together with structural genes for a sucrose hydrolase (invertase) CscA, for a d-fructokinase CscK, and for a transport system CscB. Based on DNA sequencing studies, this last codes for a hydrophobic protein of 415 amino acids. CscB is closely related to the -galactoside transport system LacY (31.2% identical residues) and a raffinose transport system RafB (32,3% identical residues) of the enteric bacteria, both of the proton symport type. A two-dimensional model common to the three transport proteins, which is based on the integrated consensus sequence, will be discussed.     

Protein-protein interactions in the yeastPKC1 pathway: Pkc1p interacts with a component of the MAP kinase cascade

The two-hybrid system for the identification of protein-protein interactions was used to screen for proteins that interact in vivo with theSaccharomyces cerevisiae Pkc1 protein, a homolog of mammalian protein kinase C. Four positive clones were isolated that encoded portions of the protein kinase Mkk1, which acts downstream of Pkc1p in thePKC1-mediated signalling pathway. Subsequently, Pkc1p and the otherPKC1 pathway components encoding members of a MAP kinase cascade, Bck1p (a MEKK), Mkk1p, Mkk2p (two functionally homologous MEKs), and Mpk1p (a MAP kinase), were tested pairwise for interaction in the two-hybrid assay. Pkc1p interacted specifically with small N-terminal deletions of Mkk1p, and no interaction between Pkc1p and any of the other known pathway components could be detected. Interaction between Pkc1p and Mkk1p, however, was found to be independent of Mkk1p kinase activity. Bck1p was also found to interact with Mkk1p and Mkk2p, and the interaction required only the predicted C-terminal catalytic domain of Mkk1p. Furthermore, we detected protein-protein interactions between two Bck1p molecules via their N-terminal regions. Finally, Mkk2p and Mpk1p also interacted in the two-hybrid assay. These results suggest that the members of thePKC1-mediated MAP kinase cascade form a complex in vivo and that Pkc1p is capable of directly interacting with at least one component of this pathway.     

Costs of mass-producing the root weevil, Mogulones cruciger, a biological control agent for houndstongue (Cynoglossum officinale L.)

The cost of rearing the root-feeding weevil, Mogulones cruciger Herbst, to control the invasive weed houndstongue (Cynoglossum officinale L.) was determined for two managed production methods. Production in an insectary setting provides control over rearing and all adult weevils that emerge can be collected, but required facility investment and high labor input. Mass-rearing in a managed ‘field crop’ setting required less facilities and labor while the insects were multiplying, but capture of the emerged adults was challenging and labor intensive. Estimated per adult weevil production costs were $CDN 2.65 for the insectary approach, and from $CDN 0.10 to $CDN 0.14 for mass-rearing in the managed field crop setting. Even though collection of adult weevils in the field crop production system was challenging, commercial production of M. cruciger should consider use of this mass-rearing method because of its lower cost.     

一种由粉红粘帚霉引起的青稞根腐病

[背景]根腐病在青稞生产中的危害日趋严重,阻碍了青稞根腐病的有效防控及青海省青稞产业的发展。然而人们对青稞根腐病的研究甚少且病原菌不详。[目的]明确青稞根腐病发生的危害、病原及致病性,为青稞根腐病的防控提供理论依据。[方法]采用常规的组织分离法分离青稞根腐病病原,通过形态鉴定与分子鉴定结合的方法对病原进行鉴定,并采用烧杯水琼脂法测定其致病性。[结果]共分离得到4株青稞根腐病病原菌,鉴定为Clonostachys rosea,有较强的致病性且致病性差异显著,经柯赫氏法则验证为青稞根腐病病原菌,并且是一种新的青稞根腐病病原,该类根腐病也是一种新的根腐类病害,在国内外属首次发现。[结论]Clonostachys rosea可引起青稞根腐病且致病性强。