Histopathological findings in farmed rainbow trout (Oncorhynchus mykiss) naturally infected with 3 different Aeromonas species

This study describes the macroscopic and microscopic lesions in farmed rainbow trout (Oncorhynchus mykiss) naturally infected with genetically identified Aeromonas salmonicida, A. hydrophila, and A. veronii species. The genus Aeromonas includes bacteria that naturally inhabit both waterways and organisms. At least 27 Aeromonas species have been identified to date, some of which can cause significant economic losses in aquaculture. As up to 68.8% of Aeromonas isolates may be misidentified in routine biochemical and phenotypic tests, however, reported cases of Aeromonas infection in fish may be wrongly identified. Our findings confirmed that the 3 Aeromonas species studied are associated with septicemia and dermal lesions in rainbow trout.     

Brucella intracellular life: from invasion to intracellular replication

Brucella organisms are pathogens that ultimate goal is to propagate in their preferred niche, the cell. Upon cell contact the bacteria is internalized via receptor molecules by activating small GTPases of the Rho subfamily and by a moderate recruitment of actin filaments. Once inside cells, Brucella localizes in early phagosomes, where it avoids fusion with late endosomes and lysosomes. These early events require the control of Rab small GTPases, and cytokines such as the G-CSF. Then, the bacterium redirects its trafficking to autophagosomes and finally reaches the endoplasmic reticulum, where it extensively replicates. Some of the bacterial molecular determinants involved in the internalization and early events after ingestion are controlled by the BvrS/BvrR two component regulatory system, whereas the intracellular trafficking beyond this early compartments are controlled by the VirB type IV secretion system. Once inside the endoplasmic reticulum, Brucella extensively replicates without restricting basic cellular functions or inducing obvious damage to cells. The integrity of Brucella LPS on the bacterial surface is one of the required factors for Brucella intracellular survival, and therefore for virulence.     

Comparison of three types of adhesives in attaching coral transplants to clam shell substrates

• 1. Nubbins from 12 coral species were transplanted onto dead giant clam shells at three sites in a lagoon near Bolinao, north‐western Philippines. Transplants were attached using three types of adhesives: cyanoacrylate glue (SG), epoxy putty (EP) and marine epoxy (ME) and were monitored over five months for detachment, in situ mortality and natural self‐attachment by tissue growth.
• 2. Corals attached with SG showed a significantly higher rate of detachment (logrank test) than those attached with either EP or ME (P<0.001 in each case). Also, those attached with EP showed a higher rate of detachment than corals attached with ME (logrank=6.46, P<0.05).
• 3. ANOVA (P>0.05) and survival analysis (logrank=2.85, P>0.05) showed no among‐adhesives differences in in situ mortality. Among the species used, Porites cylindrica, Pavona frondifera, Heliopora coerulea and Porites rus had the highest survivorship with 82%, 80%, 76% and 73%, respectively, of nubbins alive at the end of the experiment and also displayed the highest cumulative rates of self‐attachment. Species with the lowest survival rates included Acropora muricata and four pocilloporids.
• 4. Although there was no significant difference in the number of self‐attaching transplants among the adhesives, corals transplanted using ME and EP self‐attached earlier than those transplanted using SG (logrank=11.19, P<0.01).
• 5. Results from this study highlight the importance of selecting the appropriate means of attaching corals to substrates and carefully considering species‐specific traits of candidate corals for transplantation.

Copyright © 2008 John Wiley & Sons, Ltd.     

Rough vaccines in animal brucellosis: structural and genetic basis and present status

Brucellosis control and eradication requires serological tests and vaccines. Effective classical vaccines (S19 in cattle and Rev 1 in small ruminants), however, induce antibodies to the O-polysaccharide of the lipopolysaccharide which may be difficult to distinguish from those resulting from infection and may thus complicate diagnosis. Rough attenuated mutants lack the O-polysaccharide and would solve this problem if eliciting protective immunity; the empirically obtained rough mutants 45/20 and RB51 have been used as vaccines. Strain 45/20 is reportedly unstable and it is not presently used. RB51 is increasingly used instead of S19 in some countries but it is rifampicin resistant and its effectiveness is controversial. Some controlled experiments have found good or absolute protection in adult cattle vaccinated orally (full dose) or subcutaneously (reduced dose) and in one field experiment, RB51 was reported to afford absolute protection to calves and to perform better than S19. Controlled experiments in calves, however, have shown reduced doses of RB51 to be ineffective, full doses only partially effective, and RB51 less effective than S19 against severe challenges. Moreover, other observations suggest that RB51 is ineffective when prevalence is high. RB51 is not useful in sheep and evidence in goats is preliminary and contradictory. Rough mutants obtained by molecular biology methods on the knowledge of the genetics and structure of Brucella lipopolysaccharide may offer alternatives. The B. abortus manBcore (rfbK) mutant seems promising in cattle, and analyses in mice suggest that mutations affecting only the O-polysaccharide result in better vaccines than those affecting both core and O-polysaccharide. Possible uses of rough vaccines also include boosting immunity by revaccination but solid evidence on its effectiveness, safety and practicality is not available.     

Relationship of European persimmon (Diospyros kaki Thunb.) cultivars to Asian cultivars, characterized using AFLPs

Sixty one persimmon (Diospyros kaki Thunb.) selections, including 17 Italian, 11 Spanish, 13 Japanese, six Korean, five Chinese, one Israeli, and eight of unknown origin, were evaluated for genetic differences by AFLP analysis. Relationships among cultivars were evaluated by UPGMA clustering, Neighbor Joining, and MultiDimensional Scaling. While similarities among groups were generally less than 0.60, both UPGMA and Neighbor Joining separated European and Asian cultivars. Spanish and Italian cultivars were not separated by any of the analyses, suggesting that they share a common gene pool, while Japanese, Chinese and Korean cultivars formed distinct clusters. Diversity within groups was greater than diversity between groups. Most cultivars were quite polymorphic (only 0.60–0.80 similarity between cultivars). In addition, the presence of several Japanese cultivars in the European group and a group of European cultivars nested between Chinese and Korean groups suggest that similar, but different progenitors were used in the development of the present European cultivars. ‘Kaki Tipo’ selections from different sources were clearly different by AFLP analysis, indicating that they are separate cultivars.     

Tuberization of the potato plant under non-inducing conditions

Zusammenfassung Der Einfluss einer verl?ngerten Inkubation der Mutterknolle auf den Beginn der Knollenbildung bei Pflanzen, die unter nicht induzierenden Bedingungen gewachsen waren, wurde untersucht. Von 14 Wochen nach Beginn der Inkubation (8 Wochen nach dem Keimen) an wurden in 5 Zeitabst?nden von je 30 Tagen Muster von 30 Knollenstücken der SorteKemebec entnommen. Sie wurden in T?pfe mit einer Mischung von gleichen Teilen Erde und Vermiculit ausgepflanzt. Die Pflanzen wurden in Klimakammern unter künstlichem Licht bei 24 C · 1 gehalten bei einer Belichtungsdauer von 8 Stunden für die eine H?lfte und 16 Stunden für die andere H?lfte der Pflanzen. Die Pflanzen aller fünf Verfahren bildeten unter Kurztag-Bedingungen Knollen. Unter Langtag-Bedingungen entwickelten die Pflanzen der ersten drei Verfahren keine Knollen, wohl aber jene der verbleibenden zwei Verfhren (tabelle). Daraus schloss man, dass die Zeitspanne zwischen dem Auspflanzen und der Knollenbildung allm?hlich kürzer wird, wenn die Kartoffelknollen in einem fortgeschrittenen Stadium der Inkubation gepflanzt werden; auch die H?he der Pflanzen nimmt unter Langtag-Bedingungen st?ndig ab. Knollen, die nicht ein bestimmtes Inkubationsstadium erreicht haben, bilden unter nicht induzierenden Bedingungen (1., 2. und 3. Pflanztermin unter Langtag) keine Knollen. Jene, die diese kritische Schwelle überschritten haben (4. und 5. Pflanztermin), erzeugen Knollen.

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Résumé On a recherché l'influence d'une incubation prolongée du tubercule-mère sur l'initiation des tubercules, les plantes poussant dans des conditions non inductives. A partir de 14 semaines après le commencement de l'incubation (8 semaines à partir de la germination), des séries de 30 fragments de tubercules de la variétéKennebec étaient prélevés à cinq intervalles de 30 jours et plantés en pots contenant un mélange en parties égales de sol et de vermiculite. Les plantes poussaient dans des chambres conditionnées sous lumière artificielle à 24 C▴ 1, une moitié des plantes recevant 8 heures et l'autre 16 heures de lumière. Sous des conditions de jours courts, les plantes des cinq traitements tubérisaient. Sous des conditions de jours longs, les plantes des trois premiers traitements ne tubérisaient pas, et celles des deux traitements restants mourraient. La conclusion est que si l'on plante des tubercules de pomme de terre dans un stade avancé d'incubation, la période entre la plantation et la tubérisation décroit progressivement; de même la hauteur des tiges décroit progressivement sous des conditions de jours longs. Les tubercules qui n'ont pas atteint un stade déterminé d'incubation ne tubérisent pas sous des conditions non induetives (ler, 2e et 3e plantations sous jours longs). Ceux qui ont passé ce seuil (4e et 5e plantations) tubérisent.

     

Characterization of under-utilized fruits by molecular markers. A case study of loquat

The usefulness of RAPD markers for genotyping a minor fruit species such as loquat has been tested in order to assess their ability for identifying accessions and to provide a set of markers suitable for use by different groups of scientists and curators. Twenty-nine polymorphic markers selected from a previous study of 33 accessions were tested in 46 new accessions added to the collection. Using the same PCR standard conditions, only 20 markers out of 29 selected in the previous study gave consistent amplifications in the new set of plant material. The rest required optimization of reaction conditions. This fact pointed out that RAPD markers were sensitive to the experimental conditions, hence a standard technique did not guarantee reproducibility. To overcome this problem markers for plant fingerprinting should be selected after comparison across accession sets. Only those markers reproducible with different sampling and checked in several sets of accessions are suitable for germplasm fingerprinting. In the present study we propose 31 RAPDs for fingerprinting loquat that accomplished these characteristics.The markers obtained were sufficient for determining origin and relationships of cultivars, for identifying synonyms and derived varieties from bud-sports. All bud sports were identical for all RAPDs selected.     

Induction of variability in chimeric Aster cordifolius `White Elegans' through somaclonal variation

A protocol was developed for plant regeneration from leaf explants (laminaand petiole) of Aster cordifolius cultivar `White Elegans', which ischimeric for chromosome number and the ligulate flower colour. Explantswere cultured on MS or Gamborg's B5 medium in presence of IAA, 2,4-Dand BAP, alone or in combination. The highest frequency of shootregeneration was obtained on B5 medium supplemented with 2,4-D(0.09 mg l^-1) and BAP (1.8 mg l^-1). After the invitro rooting stage and acclimatization, 90% of plantlets survived in thegreenhouse. The regeneration process occurred by indirect organogenesisas shown by cyto-histology. The regenerants maintained the originalchimerism for chromosome number. Pink flowers instead of theexpected white ones were obtained in all regenerants. Ahypothesis for the uniform colour change is discussed. Variations weredetected in the regenerant generation (R₀) for quantitative characterssuch as capitulum and disc diameters, and number of ligulate flowers. SinceAster can be vegetatively propagated, selection of variants in R₀can provide interesting material for breeding purposes.     

Identification of <Emphasis Type="BoldItalic">Pasteurella multocida</Emphasis> capsular types isolated from rabbits and other domestic animals in Mexico with respiratory diseases

Pasteurella multocida is the causative agent of pasteurellosis, a major disease in most domestic animals and livestock. In this study, a total of 34 isolates of P. multocida from rabbits and other domestic animals from Mexico with respiratory diseases underwent polymerase chain reaction-based capsular typing. One sheep isolate was found to belong to capsular serogroup D, whereas the rest of the rabbit, sheep, cattle, pig, goat, and duck isolates belonged to capsular serogroup A of P. multocida. This is the first report of capsular type A in P. multocida isolates from rabbits and duck origin in Mexico.