Extracellular proteins of Clostridium chauvoei are protective in a mouse model

The anaerobic bacillus Clostridium chauvoei is the causative agent of blackleg, a lethal disease that has an important impact on the sheep and cattle industry worldwide. Immunity to C. chauvoei is considered to be mainly anticellular, and for this reason there is scarce information about the immunogenicity of extracellular proteins. In this work variations in protein profiles, immune response by ELISA and protective capacity of culture supernatants of three C. chauvoei strains, collected at different growth phases, are reported. Sera raised against extracellular antigens also recognised cellular antigens of the same molecular masses. Partially purified cell-free supernatants and those concentrated 10 times by ultrafiltration (C-CFS), obtained at the early stationary phase of growth, induced a strong immunoprotective response, even at low doses, that was more marked for C. chauvoei strain ATCC 10092 (p < or = 0.05). With C-CFS formulations, a clear relationship was observed between IgG titres, protective capacity and concentration of the antigen doses, indicating a specific immune response.     

Spermatocrit and spermatozoa density in Atlantic cod (Gadus morhua): correlation and variation during the spawning season

The utility of spermatocrit (the proportion of solid packed material in semen after centrifugation) as an indicator of spermatozoa density and male spawning stage was tested in Atlantic cod (Gadus morhua). Semen was collected from captive male cod over three spawning seasons. Spermatocrit was positively and significantly correlated with spermatozoa density measured with a Coulter counter (Multisizer), but not with spermatozoa counts in a haemacytometer. Spermatocrit increased significantly as the spawning season progressed. However, day of season explained only 35% of the variation because spermatocrit varied among individual males. Spermatozoa size remained unchanged throughout the sampling period and was not correlated with spermatocrit, indicating that variation in spermatocrit was due to variation in spermatozoa number and not their size. Spermatocrit is a good estimator of sperm density but is not reliable as indicator of spawning stage because of the variation among individual males.     

Ionic composition and osmolality of paddlefish (Polyodon spathula, Acipenseriformes) seminal fluid

Changes in ionic composition as Na⁺,K⁺, Ca²⁺ and Mg²⁺, osmolality inseminal fluid, percentage of motile spermatozoaand velocity were investigated in response toCPP and different dosage of LHRHa. The lowestvelocity of sperm was observed after use CPPtreatment. The velocity of spermatozoa,significant main effect of the treatment(P < 0.0001) and the time of sperm collection(P < 0.0104) were evaluated. The osmolality ofseminal fluid was different betweenexperimental groups of LHRHa (48.0–62.7mOsmol.kg^–1) and CPP (33.0–46.3mOsmol.kg^–1) treatments. The osmolalitywas significantly higher on the first day andone-half, then declined on day three, rangingfrom 33.0 to 62.7 mOsmol.kg^–1. Analysisof variance showed significant main effects ofthe treatment (P < 0.0001) and the time ofsperm collection (P < 0.0002) on the osmolalityof seminal fluid. The level of Na⁺ andK⁺ ion was different between experimentalgroups of LHRHa and CPP treatment. The highestconcentration of 11.11 mmol.l^–1 wasobserved at Na⁺ ion. Then theconcentrations declined on the level 1.56, 0.52and 0.36 mmol.l^–1 for K⁺, Ca²⁺and Mg²⁺ ions, respectively. There werehighly positive correlations between osmolalityof seminal fluid and dosage of LHRHa treatment(r = 0.84), velocity of spermatozoa andosmolality of seminal fluid (r = 0.57) andosmolality of seminal fluid and Na⁺concentration at seminal fluid (r = 0.70).Injection with LHRHa increased quality of spermas velocity of sperm, level of Na⁺,K⁺ and osmolality at seminal fluidcompared to CPP treatments.     

Development of a method for live octopus (Octopus vulgaris) transportation for long distance at high densities

The development of new octopus‐based products with a growing economic value ensures the commercial interest of this species, making live octopus export an activity of great interest. The aim of this study was to develop a method for long‐distance transportation of live octopus at high densities. The system was composed by 220‐L tanks with cooling and aeration, where the animals were kept separated from each others. The water temperature was maintained at 10°C, after a decreasing of 1°C/hr. Live octopus transportation was tested for 48 hr at two densities: 50 kg/m³ and 100 kg/m³. During this period, water parameters were monitored. Stress response was evaluated through the analysis of haemolymph, muscle and brain tissues. No mortality was registered after 48 hr for both treatments. In all trials, water quality remained within the normal limits in both densities; there was, however, a significative increase of ammonia levels in the water. Ammonia, dopamine and Hsp70 levels were analysed in the beginning and at the end of the experiment for both densities; however, no significant differences were found among them. In general, this system seems to be a viable solution for live octopus 48‐hr transportation at a density of 100 kg/m³.     

Improved husbandry to control an outbreak of rainbow trout fry syndrome caused by infection with Flavobacterium psychrophilum

CASE DESCRIPTION: A cohort of 35,200, 13-week-old, female rainbow trout at a fish farm was evaluated because of a 2-week history of anorexia and lethargy and a mortality rate of approximately 100 fish/d. CLINICAL FINDINGS: Affected fish were lethargic and thin and had disequilibrium, bilateral exophthalmia, pale red gills and kidneys, red-tinged coelomic fluid, and pale brown livers. Some fish were differentially pigmented bilaterally. The presumptive diagnosis was bacterial or viral septicemia. The definitive diagnosis was rainbow trout fry syndrome caused by infection with Flavobacterium psychrophilum. TREATMENT AND OUTCOME: A strategy for controlling the outbreak based on reducing pathogen numbers in affected tanks and reducing pathogen spread among tanks was developed. The option of treating with antimicrobial-medicated feed was discussed with the farmer, but was declined. After changes were made, mortality rate declined quickly, with no more deaths within 10 days after the initial farm visit. CLINICAL RELEVANCE: Bacterial coldwater disease is the most common manifestation of infection with F psychrophilum in fingerling and adult rainbow trout. However, the organism can also cause rainbow trout fry syndrome. This condition should be included on a list of differential diagnoses for septicemia in hatchery-reared rainbow trout fry.     

First approach to studying the genetics of the meagre (Argyrosomus regius; Asso, 1801) using three multigene families

The meagre (Argyrosomus regius) is a commercially important species for fisheries, and aquaculture production has been increasing in recent years. However, this growing importance has not been matched by increased genetic knowledge of the species. For this reason, an initial approach has been made to understanding the genetics of the meagre, using three particular multigene families (45S and 5S ribosomal DNA and U2 snRNA gene) as both molecular and cytogenetic markers. Only one type of 5S rDNA and only one ITS‐1 (from 45S rDNA) were obtained, and these therefore could provide the basis for the development of genetic markers for this species. However, the U2 snRNA gene produced a multi‐band electrophoretic pattern, and some of these bands were demonstrated to be linked with the U1 snRNA gene. This linkage could also provide a good species‐specific marker which could be useful for tracing the evolutionary history of the Sciaenidae family. In this study, we have described for the first time the karyotype of the meagre: this is composed of 48 acrocentric chromosomes. Each of the three gene families were localized on different chromosome pairs, although the U2 snRNA gene probe was also located scattered throughout the genome of the meagre, as expected by the amplification pattern.     

Progeny Production of the Copepods Pseudodiaptomus euryhalinus and Tisbe monozota in Monospecific and Mixed Cultures

The objective of this research was to culture the calanoid copepod Pseudodiaptomus euryhalinus, Johnson 1939, and the harpacticoid Tisbe monozota, Bowman, 1962, in monospecific and combined cultures (P. euryhalinus : T. monozota at 1:1, 2:1, and 1:2 starting ratios), and to compare the nauplii, copepodite, and adult production. Mean total production ranged from 740.3 ± 137.5 organisms/L (P. euryhalinus) to 884.3 ± 489.7 organisms/L (T. monozota) for monospecific cultures. The 1:1 ratio mixed cultures gave 780.4 ± 155.8 organisms/L, those with the 2:1 and 1:2 P. euryhalinus and T. monozota starting ratios produced 710.1 ± 195.2 and 799.7 ± 232.5 organisms/L, respectively, and there were no significant difference among treatments. All mixed cultures gave significantly lower copepodite productions than the monocultures of each species. In addition, the tendency to a decreased progeny production of the initial females of T. monozota indicates that the outcome of long‐term mixed cultures would production either a high dominance of P. euryhalinus, or monocultures of this species .