Different expressions of trypsin and chymotrypsin in relation to growth in Atlantic salmon (Salmo salar L.)

The expressions of trypsin and chymotrypsin in the pyloric caeca of Atlantic salmon (Salmo salar L.) were studied in three experiments. Two internal (trypsin phenotypes, life stages) and three common external factors (starvation, feeding, temperatures) influencing growth rates were varied. Growth was stimulated by increased temperature and higher feeding rate, and it was depressed during starvation. The interaction between trypsin phenotype and start-feeding temperature affected specific activity of trypsin, but not of chymotrypsin. Trypsin specific activity and the activity ratio of trypsin to chymotrypsin (T/C ratio) increased when growth was promoted. Chymotrypsin specific activity, on the other hand, increased when there was a reduction in growth rate whereas fish with higher growth had higher chymotrypsin specific activity resulting in lower T/C ratio value. During a rapid growth phase, trypsin specific activity did not correlate with chymotrypsin specific activity. On the other hand, a relationship between specific activities of trypsin and chymotrypsin could be observed when growth declined, such as during food deprivation. Trypsin is the sensitive key protease under conditions favouring growth and genetically and environmentally affected, while chymotrypsin plays a major role when growth is limited or depressed. Trypsin specific activity and the T/C ratio value are shown to be important factors in the digestion process affecting growth rate, and could be applicable as indicators for growth studies of fish in captive cultures and in the wild, especially when food consumption rate cannot be measured

A summer heat wave decreases the immunocompetence of the mesograzer, Idotea baltica

Extreme events associated with global change will impose increasing stress on coastal organisms. How strong biological interactions such as the host–parasite arms-race are modulated by environmental change is largely unknown. The immune system of invertebrates, in particular phagocytosis and phenoloxidase activity response are key defence mechanisms against parasites, yet they may be sensitive to environmental perturbations. We here simulated an extreme event that mimicked the European heat wave in 2003 to investigate the effect of environmental change on the immunocompetence of the mesograzer Idotea baltica. Unlike earlier studies, our experiment aimed at simulation of the natural situation as closely as possible by using long acclimation, a slow increase in temperature and a natural community setting including the animals’ providence with natural food sources (Zostera marina and Fucus vesiculosus). Our results demonstrate that a simulated heat wave results in decreased immunocompetence of the mesograzer Idotea baltica, in particular a drop of phagocytosis by 50%. This suggests that global change has the potential to significantly affect host–parasite interactions

Histo-Blood Group Antigens Act as Attachment Factors of Rabbit Hemorrhagic Disease Virus Infection in a Virus Strain-Dependent Manner

Rabbit Hemorrhagic disease virus (RHDV), a calicivirus of the Lagovirus genus, and responsible for rabbit hemorrhagic disease (RHD), kills rabbits between 48 to 72 hours post infection with mortality rates as high as 50–90%. Caliciviruses, including noroviruses and RHDV, have been shown to bind histo-blood group antigens (HBGA) and human non-secretor individuals lacking ABH antigens in epithelia have been found to be resistant to norovirus infection. RHDV virus-like particles have previously been shown to bind the H type 2 and A antigens. In this study we present a comprehensive assessment of the strain-specific binding patterns of different RHDV isolates to HBGAs. We characterized the HBGA expression in the duodenum of wild and domestic rabbits by mass spectrometry and relative quantification of A, B and H type 2 expression. A detailed binding analysis of a range of RHDV strains, to synthetic sugars and human red blood cells, as well as to rabbit duodenum, a likely gastrointestinal site for viral entrance was performed. Enzymatic cleavage of HBGA epitopes confirmed binding specificity. Binding was observed to blood group B, A and H type 2 epitopes in a strain-dependent manner with slight differences in specificity for A, B or H epitopes allowing RHDV strains to preferentially recognize different subgroups of animals. Strains related to the earliest described RHDV outbreak were not able to bind A, whereas all other genotypes have acquired A binding. In an experimental infection study, rabbits lacking the correct HBGA ligands were resistant to lethal RHDV infection at low challenge doses. Similarly, survivors of outbreaks in wild populations showed increased frequency of weak binding phenotypes, indicating selection for host resistance depending on the strain circulating in the population. HBGAs thus act as attachment factors facilitating infection, while their polymorphism of expression could contribute to generate genetic resistance to RHDV at the population level

Enteric coating of granules containing the probiotic Lactobacillus acidophilus

In the present study, a capsule formulation composed of enteric coated granules of Lactobacillus acidophilus ATCC 4962 was developed using Eudragit L30D-55 as enteric polymer. Optimization of the capsule formulation was achieved with a maximum viable cell count after 2 h of incubation in acid medium and disintegration time of 1 h in buffer pH 6.8. The amount of Eudragit L30D-55 in the capsules correlated with gastric juice resistance. The best protective qualities against artificial gastric juice were observed when capsules were prepared from granules composed of L. acidophilus, corn starch, lactose monohydrate, polyvinylpyrrolidone and coated with 12.5 % (m/V) of Eudragit L30D-55. Capsule formulation of L. acidophilus in edible broth medium suspension serves as a cheap alternative to the expensive freeze-drying procedure for preparing L. acidophilus. In addition, the enteric coating using Eudragit L30D-55 could protect probiotics from the acidic gastric environment and enhance the bioactivity of probiotics along with replacement of pathogenic microbes in human intestine

Digestive enzymes in the ontogenetic stages of the southern king crab, Lithodes santolla

The early ontogenetic stages of the sub-Antarctic king crab Lithodes santolla were analyzed for the presence and activities of a set of important digestive enzymes. The eggs and non-feeding larvae (zoea I-III, megalopa) showed high activities of esterases, phosphatases, and exopeptidases indicating the enzymatic ability to utilize endogeneous yolk reserves. SDS-PAGE showed a continuous decrease of proteins or proteids in the range of 59–81 kDa during ontogenetic development from the eggs through the zoeal stages to the first juvenile crab stage, CI. This reduction reflects the degradation of storage compounds during lecithotrophic larval development. Activities of the endopeptidases, trypsin and chymotrypsin, were low in eggs and larvae but increased significantly in the first juvenile crab stage. These enzymes typically facilitate the first steps of proteolysis in the extra-cellular spaces of the midgut gland and in the stomach. Their scarcity indicates that the larvae of L. santolla are physiologically not prepared to digest external food. This ability seems to appear first in the CI stage. Extracts of juvenile midgut glands and the gastric fluids of adults showed high activities of a variety of digestive enzymes including phosphatases, carbohydrases, as well as endo- and exopeptidases. High activities of digestive enzymes in adults may compensate for scarce food supply and rate-limiting low temperatures in the predominantly sub-Antarctic habitats of L. santolla

Colour or no coulour in the juvenile shell of the black lip pearl oyster, Pinctada margaritifera ?

International audiencePinctada margaritifera mollusc is cultivated in French Polynesia for the production of black pearls. For this study, the colour of juvenile samples (48 days old) was investigated in the visible range spectra (430-700 nm) using spectrophotometry. A first measurement was done with the soft parts still inside the shell (entire animal). Then, the soft parts were removed in order to do a second measurement on the growing edge of the shell. Comparison of the two measurements shows that the estimation of the living animal colour with unaided eye is strongly influenced by the colour of the soft parts. The use of the International Commission on Illumination (ISI) chromaticity diagram shows that at this growth stage, the shells are "white"; i.e. present no absorptions to the visible part. Multivariate statistical analyses based on the intensities of 6 wavelengths show that the shell colour is less variable than the colours of the entire animals. Wavelength intensities of the shells are similar, so no colour trend is visible. On the other hand, the colours of the entire animals are darker, and more variable. At this growth stage, the shell colour is not predictable for a potential selection of receiver or donor

Colour or no coulour in the juvenile shell of the black lip pearl oyster, Pinctada margaritifera ?

International audiencePinctada margaritifera mollusc is cultivated in French Polynesia for the production of black pearls. For this study, the colour of juvenile samples (48 days old) was investigated in the visible range spectra (430-700 nm) using spectrophotometry. A first measurement was done with the soft parts still inside the shell (entire animal). Then, the soft parts were removed in order to do a second measurement on the growing edge of the shell. Comparison of the two measurements shows that the estimation of the living animal colour with unaided eye is strongly influenced by the colour of the soft parts. The use of the International Commission on Illumination (ISI) chromaticity diagram shows that at this growth stage, the shells are "white"; i.e. present no absorptions to the visible part. Multivariate statistical analyses based on the intensities of 6 wavelengths show that the shell colour is less variable than the colours of the entire animals. Wavelength intensities of the shells are similar, so no colour trend is visible. On the other hand, the colours of the entire animals are darker, and more variable. At this growth stage, the shell colour is not predictable for a potential selection of receiver or donor

Rapid and sensitive PCR detection of Vibrio penaeicida, the putative etiological agent of Syndrome 93 in New Caledonia

Experimental infections of Penaeus (Litopenaeus) stylirostris were performed with a Vibrio penaeicida strain (AM101) isolated in New Caledonia from Syndrome 93 diseased shrimp. Cumulative mortalities resulting from intramuscular injection or immersion of shrimp in bacterial suspensions demonstrated high virulence for this bacterial strain and suggested that V. penaeicida could be the etiological agent of Syndrome 93. The median lethal dose (LD50) for AM101 was 1.3 x 104 CFU (colony forming units) ml-1 by immersion and less than 5 CFU shrimp-1 by intramuscular challenge, with mortality outbreaks at 48 and 22 h after challenge, respectively. A polymerase chain reaction (PCR) detection assay using a primer set designed from the 16S ribosomal RNA gene of V. penaeicida was developed. It gave an expected amplicon of approximately 310 bp in ethidium bromide-stained agarose gels. The specificity of these primers was assessed with different Vibrio species. Furthermore, DNA extracted by the ChelexTM method could be used to detect fewer than 20 cultured Vibrio cells in seawater or shrimp hemolymph by this assay. It appears to be a reliable screening method for detecting V. penaeicida in shrimp and from the aquatic environment

Hemolymph metabolic variables and immune response in Litopenaeus setiferus adult males: the effect of acclimation

Massive nauplii production in Litopenaeus setiferus by means of natural matings has not been reported. The main reason is the low spermatophore attachment rate that has been associated with male sterilization because of male reproductive tract degenerative and the male reproductive melanization syndromes (MRTDS and MRMS). This information indicated both syndromes could be related to the captivity and management stress that affects the immune system and the physiological state of shrimp. We used some blood metabolic variables, sperm quality, and immune response as indicators of captivity stress in adult males of L. setiferus. A comparison between freshly captured shrimp with shrimp maintained in the laboratory for 7 days at two temperatures were examined. Glucose and calcium were not different between fresh and acclimated shrimp at either temperature (P > 0.05). A reduction in triacylglycerol, proteins, and cholesterol was observed in acclimated shrimp in comparison with base line shrimp (P < 0.05). Lactate was reduced only in shrimp acclimated at the lower temperature. Sperm quality was not significantly different between base line samples and acclimated shrimp. The immune system was altered in acclimated shrimp at both temperatures. A reduction in total haemocytes, granular cells, and semigranullar cells was measured in acclimated shrimp in comparison with base-line shrimp. In contrast, a higher phenoloxidase activity (proPO) was observed in acclimated shrimp, indicating that regulatory mechanisms of immune system of those shrimp were altered by captivity conditions. The blood metabolic variables indicated that captive shrimp were affected nutritionally more than physiologically. The immune response showed the nutritional effect or another management factor reduced the cellular defenses and altered the molecular mechanisms associated with melanin production. This could be related to the melanization syndrome observed in previous studies. A new sequence to explain the appearance of the male reproductive shrimp syndromes of L. setiferus was proposed. (C) 2001 Elsevier Science B.V. All rights reserved