Cellular immunophenotypic profile in the splenic compartment during canine visceral leishmaniasis

To determine the role of the spleen in the pathogenesis of canine visceral leishmaniasis (CVL), we analyzed cellular immunophenotypic profiles of 52 dogs naturally infected with Leishmania infantum, clinically classified as follows: asymptomatic dogs-I (AD-I), seronegative/PCR+; asymptomatic dogs-II (AD-II), seropositive/PCR+; oligosymptomatic dogs (OD) and symptomatic dogs (SD). Seven non-infected dogs (CD) were included as a control group. AD-II presented higher levels of CD8+ T splenocytes and lower TCD4+/TCD8+ ratio in comparison with CD. OD and SD showed lower percentages of CD21+ as compared with AD-II. All seropositive dogs presented lower levels of CD45RA+ than CD. Regardless of the stimuli used, the proliferation index from splenocytes in vitro was inversely correlated with clinical status. After LSA stimulation, there was a higher percentage of specific CD8+ T in AD-II than CD and non-stimulated culture. In contrast, splenocytes from SD under in vitro LSA stimulation induced decreased MHC-II+ expression in comparison with all groups, and non-stimulated culture. In conclusion, the role of CD8+ T splenocytes seems to be important for an effective immunological response, a hallmark of asymptomatic CVL, whereas the pronounced loss of MHC-II expression upon LSA stimulation is a biomarker of symptomatic CVL.     

Long-term Electrocardiography Recording with Holter Monitoring in 15 Donkeys

The heart rates (HR) and rhythms of 15 healthy donkeys of various ages, both sexes, and various breeds were analyzed throughout 24-hour Holter monitoring. The animals were evaluated at rest in their daily environment without the presence of investigators causing stress, using a three-channel digital Holter recorder. Analysis revealed a maximal HR range from 62.5 to 93.7 beats/min (mean, 72.50 ± 7.51 beats/min), whereas the minimal HR ranged from 29.7 to 42.2 beats/min (mean, 34.90 ± 4.22 beats/min). The daily mean HR was 47.55 ± 3.70 beats/min. The daytime mean HR was 49.39 ± 2.77 beats/min, whereas the night time rate was 46.22 ± 4.38 beats/min. Sinus bradycardia and sinus tachycardia were detected in 7 and 10 of 15 studied donkeys, respectively. Cardiac dysrhythmias due to high vagal tone such as sinoatrial heart block and second-degree atrioventricular heart block were occasionally recorded in 1 and 3 donkeys, respectively. A higher mean HR and fewer cardiac dysrhythmias were observed in donkeys than in horses and ponies at rest. These findings could be justified by differences in the autonomic nervous system tone.     

Occurrence and present status of coastal dolphins (Delphinus delphis and Tursiops truncatus) in the eastern Ionian Sea

• 1. Boat surveys aimed at studying short‐beaked common dolphins and common bottlenose dolphins in eastern Ionian Sea coastal waters were conducted between 1993 and 2003. During 835 survey days, 24 771 km of total effort was distributed within an area of 480 km², resulting in 428 common dolphin and 235 bottlenose dolphin sightings.
• 2. Individual photo‐identification was performed extensively throughout this study, making it possible to monitor the number of animals seen in the study area each year and their long‐term residency patterns.
• 3. Common dolphins declined across the study period, from 2.18 encounters/100 km in 1997 to 0.40 encounters/100 km in 2003. In contrast, there was a relatively stable presence of bottlenose dolphins, some individuals showing high levels of site fidelity and others using the area only occasionally.
• 4. The local decline of common dolphins and the low density of bottlenose dolphins appeared to reflect the general status of these cetacean species in the wider Mediterranean region, where common dolphins were classified as endangered in the IUCN Red List in 2003.
• 5. Based on the available evidence, we infer that the present unfavourable status of common dolphins in eastern Ionian Sea coastal waters is largely a consequence of prey depletion.

Copyright © 2005 John Wiley & Sons, Ltd.     

Production of meiotic gynogenetic and triploid sea bass, Dicentrarchus labrax L.: 1. Performances, maturation and carcass quality

Meiotic gynogenetic and triploid sea bass were produced by pressure shocks according to a previously published protocol. Pressure-treated groups did not survive as well as controls during early development and larval rearing. Performances, sexual maturation and carcass quality were examined over a period of 34–45 months. At the age of 34 months, growth of the gynogenetic fish was comparable to that of the control but inferior in the triploid fish. A predominance of male fish was found within the triploid groups, while diploid and meiotic gynogenetic fish showed equal proportions of the sexes. Gonadal maturation in triploid fish was significantly impaired, particularly in the females that showed rudimentary ovaries. Triploid males exhibited primary maturation but proved to be gametically sterile. Pressure-induced triploids did not grow as well as diploids, but these results might be ascribed to specific on-growing conditions (communal rearing). The performance of gynogenetic sea bass was comparable to that of control. The superiority of diploid fish over their triploid counterparts was confirmed during the final growing period and more clearly so in females. Performances of triploids varied according to their maternal origin. Overall, striking qualitative differences between diploid and triploid fish were found at the age of 34 and 45 months, although the results varied in a gender-specific manner. A strong maternal effect was also observed. The potential advantages of triploid sea bass for aquaculture purposes are discussed.     

Purification of chinook salmon (Oncorhynchus tshawytscha) GH for receptor study

A method for the purification of chinook Salmon (Oncorhynchus tshawytscha) GH, which retains its biological activity, is described. The biological activity was investigated with an established radioreceptor assay using liver membranes from pregnant rabbits and bovine GH as standard and labelled hormone. The enrichment of the preparation was checked with electrophoresis (SDS-PAGE). Extraction and further steps were carried out using low molarity alkaline buffer (pH 8–10, M = 100 mM). Three chromatography steps were performed (Concanavalin-A sepharose, Bio-gel P60, DEAE). Ion exchange chromatography was performed under isocratic conditions (using a 50 cm column). Two isoforms (sGH1 and sGH2) were isolated. The purification yield is 0.7% compared to lyophilized pituitaries. The molecule is homogeneous in SDS-PAGE. Contamination by prolactin, gonadotrophin and corticotrophin is negligible (< 0.5%). It could be demonstrated that the biological activity of the preparation is maintained since this preparation stimulates the growth of juvenile trout (Salmo gairdneri) and binds specifically (35%) to trout liver membranes.     

Summer mortality of hatchery-produced Pacific oyster spat (Crassostrea gigas). I. Estimation of genetic parameters for survival and growth

The multidisciplinary project “MOREST” aims to improve our understanding of causes of summer mortality in Crassostrea gigas juveniles in France and to reduce its impact on oyster production. As part of the MOREST project, 43 full-sib families nested within 17 half-sib families were produced, planted out and tested in 3 sites during summer 2001 to assess to what extent genetic variability exists for this trait. A strong genetic basis was found for survival as narrow-sense heritability estimates ranged from 0.47 to 1.08, with higher values in sites where summer mortality was higher. Genetic correlations across sites were positive and very high for survival, indicating no genotype by environment interaction. In contrast, lower genetic variation was observed for growth in all sites. Finally, genetic correlations between growth and survival were low, in all sites. Selective breeding in a single site should therefore be an efficient means of improving survival in oysters less than one year old along the French Atlantic and Channel coastlines with only very limited effects on growth. As yield mostly depends on survival and growth, this approach should significantly improve harvestable yield. Possible reasons why a high genetic variance for survival appears to be maintained in wild populations are discussed.     

Quantifying and simulating carbon and nitrogen mineralization from diverse exogenous organic matters

The potential contributions of exogenous organic matters (EOMs) to soil organic C and mineral N supply depend on their C and N mineralization, which can be assessed in laboratory incubations. Such incubations are essential to calibrate decomposition models, because not all EOMs can be tested in the field. However, EOM incubations are resource-intensive. Therefore, easily measurable EOM characteristics that can be useful to predict EOM behaviour are needed. We quantified C and N mineralization during the incubation of 663 EOMs from five groups (animal manures, composts, sewage sludges, digestates and others). This represents one of the largest and diversified set of EOM incubations. The C and N mineralization varied widely between and within EOM subgroups. We simulated C and N mineralization with a simple generic decomposition model. Three calibration methods were compared. Individual EOM calibration of the model yielded good model performances, while the use of a unique parameter set per EOM subgroup decreased the model performance, and the use of two EOM characteristics to estimate model parameters gave an intermediate model performance (average RMSE-C values of 32, 99 and 65 mg C g⁻¹ added C and average RMSE-N values of 50, 126 and 110 mg N g⁻¹ added N, respectively). Because of the EOM variability, individual EOM calibration based on incubation remains the recommended method for predicting most accurately the C and N mineralization of EOMs. However, the two alternative calibration methods are sufficient for the simulation of EOMs without incubation data to obtain reasonable model performances.