Digestive enzyme activity during early larval development of the Cuban gar Atractosteus tristoechus

The ontogenesis of digestive enzymes (proteases, amylases, lipases, and phosphatases) in Cuban gar Atractosteus tristoechus was determined in larvae between 5 and 18 days after hatching (DAH). Variations in specific activities of most enzymes were related to the transition from endogenous to exogenous feeding and to the transition from the larval to the juvenile stage. Alkaline protease activity was not detected until 8 DAH in contrast to acid protease activity, which was quantifiable at 5 DAH. Acid protease activity was consistently higher than alkaline protease activity, indicating the presence of a functional stomach in the early stages of larval development. The acid protease activities of larvae and adults were compared by means of zymogram analysis. Four acid protease bands were found in adults (two more than in larvae). This result is the first time that more than one band of acid proteolytic activity has been found in Lepisosteidae. High lipase activity indicated the importance of lipid utilization, particularly during yolk-sac absorption. In contrast to the other enzymes studied, amylase activity was consistently low, probably due to the strictly carnivorous diet of gar larvae and their low capacity for carbohydrate digestion. High activities of aminopeptidase and acid and alkaline phosphatases suggest intestinal absorption. This result, together with the existence of a short gut and a lower proteolytic activity in the intestine than in the stomach, suggest that most of the proteolytic activity takes place in the stomach, while the primary function of the intestine is nutrient uptake.     

Prevalence and different characteristics of two serotypes of Streptococcus parauberis isolated from the farmed olive flounder, Paralichthys olivaceus (Temminck and Schlegel), in Korea

The prevalence of two serotypes of Streptococcus parauberis isolated from the olive flounder, Paralichthys olivaceus, was evaluated in a total of 29 isolates between 2003 and 2010 in Korea. Streptococcus parauberis isolates were divided into two serologically distinct types (serotype 1 and serotype 2), except for one strain (S1091), using an agglutination assay with rabbit antiserum, and serotype 1 was identified as the dominant type (24 of 29 isolates) in this study. To identify the characteristics of the two serotypes of S. parauberis, we conducted a biochemical test using the API 20 Strep kit, a transmission electron microscopy (TEM) assay, sequence analysis of 16S‐23S rRNA intergenic spacer region (ISR) and a pathogenicity test. In TEM, both serotypes possessed polysaccharide capsule layers around the cell surface when bacterial cells were treated with a homologous serotype of rabbit antiserum. However, we were unable to discriminate serotype‐specific biochemical characteristics and genetic characteristics of 16S‐23S rRNA ISR between the two serotypes. In the pathogenicity test, the serotype 1 strains induced significantly higher mortality than the serotype 2 strains in olive flounder when experimentally inoculated via the intraperitoneal route.     

Efficacy of furunculosis vaccines in turbot, Scophthalmus maximus (L.): evaluation of immersion, oral and injection delivery

The commercial furunculosis vaccine Aquavac Furovac 5 and an autogenous vaccine, based on the challenge strain, induced immune protection in turbot, Scophthalmus maximus (L.), as shown in challenge tests 120 days post-immunization by injection (relative percentage of survival, RPS = 72-99%). This protective effect lasted for at least 6 months post-immunization at appreciable levels (RPS = 50-52%). Neither the autogenous vaccine nor the commercial vaccine was able to induce significant levels of protection against Aeromonas salmonicida in turbot when administered by immersion. Antibody levels were high or moderate in fish vaccinated by injection with the different vaccines and very low in fish vaccinated by immersion. The field results show that delivering an oral boost after the primary vaccination by injection did not enhance protection of turbot against furunculosis and that water-based (autogenous vaccine) and oil adjuvanted (Alpha Ject 1200) vaccines administered by injection conferred similar levels of protection (RPS > 80%) in turbot.     

Dietary magnesium requirement and physiological responses of marine shrimp Litopenaeus vannamei reared in low salinity water

An 8‐week feeding experiment was conducted to determine the dietary magnesium (Mg) requirement and physiological responses of Litopenaeus vannamei in low salinity water of 2 g L^?1. Casein–gelatin‐based diets supplemented with seven levels of Mg (0, 0.4, 0.8, 1.6, 3.2, 6.4 and 8.0 g kg^?1) were fed to juvenile shrimp. Prior to the experiment, the postlarvae were gradually acclimated to the low salinity media and fed with a basal diet (0.5 g Mg kg^?1) for 2 weeks. After 8 weeks of feeding, survival ranged from 80.11% to 85.65% with no significant difference among the treatments. Hepatopancreas Mg²⁺‐ATPase and Na⁺/K⁺‐ATPase activities and muscle content of lipid and protein were not significantly affected by graded levels of Mg. The weight gain and mineral (calcium, potassium, sodium and total phosphorus) content of different tissues were significantly affected by dietary Mg levels, while there were no significant differences in ash and zinc content in tissues. The Mg content in tissues except hepatopancreas was maintained relatively constant regardless of dietary treatments. The dietary Mg requirement for optimal growth was 2.60–3.46 g Mg kg^?1 by using the polynomial regression analysis based on growth.     

Enigmatic hotspot of crayfish diversity at risk: Invasive potential of non‐indigenous crayfish if introduced to New Guinea

1. The large island of New Guinea has a rich indigenous astacofauna represented by numerous parastacids from the genus Cherax. The western half of the island is part of Indonesian territory.
2. Indonesia is known to be the main exporter of ornamental crayfish globally, and certain New Guinean species are exploited as ornamentals within the international pet trade. Moreover, one non‐indigenous species has been previously recorded being cultured in Java, Indonesia. This species, the North American Procambarus clarkii, is a vector of crayfish plague, the disease that is lethal to most parastacids. This population has already tested positive for the disease.
3. As the transport of non‐indigenous crayfish within the Indonesian territory is not restricted, their introduction to New Guinea can be expected. The Indonesian market was therefore surveyed for ornamental crayfish and their environmental suitability evaluated, as represented by temperature during the drought and rainy seasons in New Guinea.
4. Four North American and one Australian species were found advertised for sale. One of them, P. clarkii, was assessed as the most damaging species, followed by other North American species. A total ban on the culture and transport of the highest risk crayfish species in Indonesia and Papua New Guinea is recommended.

     

Detection of Flexibacter maritimus in fish tissue using nested PCR amplification

A nested polymerase chain reaction (PCR) system was developed for the detection of Flexibacter maritimus from fish tissue. The total procedure for the diagnosis of marine flexibacteriosis, from the point of DNA extraction to the electrophoretic analysis, can be performed in < 4 h. This was achieved by the combination of a short thermal cycling programme with a rapid DNA extraction procedure. The assay was extremely sensitive, capable of detecting as few as 75 cfu mg(-1) fish tissue. The accuracy of the nested PCR was confirmed under field conditions using tissue samples recovered during 1993-2002 from fish suffering marine flexibacteriosis. The nested PCR method proved to be efficient for the rapid and sensitive detection of F. maritimus from fish tissues and can be used for routine diagnosis of the disease caused by this pathogen.     

Detection of antigenic proteins expressed by lymphocystis virus as vaccine candidates in olive flounder, Paralichthys olivaceus (Temminck & Schlegel)

Although the major capsid proteins (MCPs) of lymphocystis disease virus (LCDV) have been characterized, little is known about the host-derived immune response to MCPs and other LCDV antigenic proteins. To identify antigenic proteins of LCDV that could be used as vaccine candidates in olive flounder, Paralichthys olivaceus, we analysed the viral proteins responsible for its virulence by applying immuno-proteomics. LCDV proteins were separated by one-dimensional gel electrophoresis, transferred to polyvinylidene difluoride membrane, and probed with homogeneous P. olivaceus antisera elicited by LCDV natural infection and vaccination with formalin-killed LCDV. Four immune-reactive proteins were obtained at 68-, 51-, 41- and 21 kDa using antisera collected from natural infection while two proteins at 51- and 21 kDa exhibited response to antisera from vaccinated fish, indicating that the latter two proteins have vaccine potential. Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nanoelectrospray MS/MS, the 51 and 21 kDa proteins were identified as MCP and an unknown protein, respectively.     

Identification and characterization of lactic acid bacteria isolated from rainbow trout, Oncorhynchus mykiss (Walbaum), with inhibitory activity against Lactococcus garvieae

A study was conducted to evaluate the probiotic properties of endogenous rainbow trout microbiota against pathogenic Lactococcus garvieae. A total of 335 bacterial strains were isolated from rainbow trout and screened for antagonistic activity against L. garvieae using an agar spot assay. Antagonistic strains were grouped by PCR amplification of repetitive bacterial DNA elements (rep‐PCR) and identified by 16S rRNA gene sequence analysis. The results revealed that the antagonistic strains belonged to the genera Lactobacillus, Lactococcus and Leuconostoc. Further probiotic characteristics, such as specific growth rate, doubling time, resistance to biological barriers, antibiotic resistance, hydrophobicity and production of antimicrobial substances, were also studied. These strains were able to survive low pH and high bile concentrations, showed good adherence characteristics and a broad spectrum of antibiotic resistance. The antagonistic efficacy was maintained after sterile filtration and was sensitive to proteinase K, indicating that proteinaceous extracellular inhibitory compounds were at least partially responsible for pathogen antagonism. Based on these results, these strains should be further studied to explore their probiotic effects in challenge experiments in vivo. This study shows clear evidence that the indigenous trout‐associated microbiota may provide a defensive barrier against L. garvieae.     

Effects of steroid hormones on immunoglobulin M (IgM) in rainbow trout, Oncorhynchus mykiss

The immunosuppressive effects of steroid hormones were evaluated as the response against implanted steroid hormones, cortisol (F), testosterone (T), estradiol-17 (E2), and 11- ketotestosterone (11-KT), in juvenile rainbow trout. In long term experiments (5 weeks), fish were given a single intraperitoneal implant of F or T. A clear suppressive effect of plasma IgM levels with F and T was not necessarily obtained, although mucus IgM levels were reduced corresponding to the elevated plasma steroid hormone levels. In short term experiments (1 week), intraperitoneal implantation of T, 11-KT and E2 suppressed plasma and mucus IgM levels, although the effects were not dose-dependent. When administered through diet, F and T caused a suppression of plasma IgM levels; F administration at both high and low dosages caused a significant decrease in plasma IgM levels, while only a high dose of T caused the suppression. These results suggest that sex steroid hormones, as well as F, have immunosuppressive functions in rainbow trout.     

基于神经网络预测的无线传感器网络田间射频信号路径损耗

为解决应用无线传感器网络技术监测农田信息时无法快速预测射频信号路径损耗的问题,基于神经网络理论研究了田间路径损耗与其影响因素间的关系。试验中选取915和2 470 MHz 2个载波频率,在冬小麦的不同生长阶段测量射频信号在田间各影响因素作用下的路径损耗,建立和验证基于神经网络的射频信号田间路径损耗预测模型。所建立模型模拟值与实测值的相关系数为0.92,应用建立的神经网络预测田间射频信号路径损耗并与实测值对比,最大预测误差绝对值为4.186 dB,最大预测标准差为2.759 dB,预测准确度为94.2%。所建立的BP网络可以对田间射频信号路径损耗进行预测。