Aflatoxin B1 contamination of shrimp feeds and its effect on growth and hepatopancreas of pre-adult Penaeus monodon

A survey of aflatoxin B₁ (AFLB₁) levels in commonly used commercial shrimp finisher feeds in the Philippines showed a various range of values from not detected to 120 μg kg^?1 using high-performance thin-layer chromatography. Six experimental diets were prepared to contain various levels of AFLB₁ based on survey results to determine the effects of such contamination in pre-adult shrimp Penaeus monodon (17.5 ± 0.6 g). Results showed that shrimps fed diets containing AFLB₁ greater than or equal to 73.8 μg kg^?1 gave comparatively poor growth rate and higher susceptibility to shell diseases. No AFLB₁ residues were detected in sampled whole shrimp tissues after 62 days of exposure to AFLB₁ containing diets indicating a low potential for transmission of the toxin from edible shrimp tissues to consumers. Histopathological alterations in the hepatopancreas of shrimp chronically exposed to AFLB, were observed in all samples. The degree of alterations correlated with the level of AFLB₁. Based on growth performance, pre-adult shrimps can tolerate AFLB₁ levels of up to 52.3 μg kg^?1 in the feeds although histopathological changes were already evident in the tissues of shrimps given diets with 26.5 μg kg^?1 AFLB₁.     

The induced damage in the hepatopancreas of Orchestia species after exposure to a mixture of Cu/Zn—An ultrastructural study

The hepatopancreas of crustaceans species has been recognized as an essential target organ to assess trace elements' effects. Due to its dynamic and capability of detoxifying trace metal, this organ often indicates distinct pathological disturbances. In the present work, we intend to evaluate the bioaccumulation of trace metal in three Orchestia species (Orchestia montagui, Orchestia gammarellus, and Orchestia mediterranea) living in symmetry in the banks of Bizerte lagoon (37°13′8″N 09°55′1″E) after their exposure during 14 days to a mixture of copper and zinc, and to highlight the effect of these metals on their hepatopancreas ultrastructure using transmission electron microscopy. At the end of the experiment, results showed that the mortality and the body mass varied according to the used nominal concentrations. Significant alterations were noted in all the treatment groups. The degree of these alterations depends on the used concentration, and they are represented especially by the cells remoteness and the border lyses, the reduction of the nuclear volume, the increase in the cytoplasm density with the presence of trace metal in the nucleus as well as in the vacuole, the disorganization and the destruction of microvilli, the condensation of the majority of cellular organelles and mitochondria swelling. Through this study, Orchestia genus could be an attractive candidate for the biochemical study of trace metal toxicity in Tunisian wetlands.     

Anionic Trypsins from Crayfish Hepatopancreas: Effects on Protein Degradation of Tail Meat

Four anionic trypsins, tentatively labeled trypsin A, B, C, and D, isolated from crayfish hepatopancreas, had high casein activity between pH 5.5 and 10.0. Temperature optima for hydrolysis of casein were 45°C for A and B, and 50°C for C and D. Trypsin D had the highest substrate turnover number (V_max) for casein reaction compared with the other trypsins at pH 6.8. Trypsins A and D had higher activities against myofibrilla and sarcoplasmic proteins from crayfish tail meat than did B and C. Results suggested that trypsins A and D are important in development of mushiness in crayfish tail meat compared with other trypsins present.     

Physiological and behavioral responses of Orchestia gammarellus (Amphipoda,Talitridae) towards trace elements contamination soil

Effects of trace elements on the hepatopancreas ultrastructure and on the locomotor activity rhythm were investigated in the amphipod talitrid Orchestia gammarellus collected from Bizerte lagoon banks situated in the north of Tunisia. Animals were exposed to a series of contaminated soil with different concentrations of cadmium and zinc. The locomotor activity rhythm was studied under constant darkness. Histopathological analysis showed that the trace elements, especially the Cd, induced significant changes in the morphology and in the ultrastructural organization of hepatopancreatic cells. The significant alterations obtained were dose dependent. Concerning the behavioral response, results revealed the presence of two components whatever the experimental conditions. In addition, a great inter-individual variability of the locomotor rhythm was observed. Patterns were in majority bimodal for the control individuals and became unimodal and multimodal when exposed under Cd and Zn, respectively. Furthermore, ultradian and circadian periods were determined. The circadian period lengthened after Zn exposure. In addition, the locomotor activity rhythm was more stable for control individuals. However, those exposed to Cd were less active.     

Trace elements bioaccumulation in Porcellionides pruinosus (Brandt, 1833) and related biomarkers of exposure

In the last years, the increase of anthropogenic activities has led to a progressive pollution of ecosystems. Oniscidean isopods are a group of terrestrial animals known as excellent bio‐indicators and bio‐accumulators, because of their capacity to accumulate contaminants and to implement appropriate strategies of detoxification. For this study, we collected Porcellionides pruinosus from two different sites in the Corinth region (Greece), a polluted site and a control site, to assess the heavy metal pollution status. We conducted an inductively coupled plasma mass spectrometry (ICP‐MS) analysis for the detection of trace elements in animals' hepatopancreas and soil; we performed also immunohistochemistry for detection of metallothioneins 1 (MT1) and heat shock protein 70 (HSP70s). The results suggest a great capacity of the species in trace elements bioaccumulation and the high degree of pollution of the area near Agioi Theodoroi where the metal pollution index (MPI) is threefolds higher than the control site, due to the presence of significantly higher concentration of arsenic, cadmium, cobalt, chromium, manganese, molybdenum, and lead found in hepatopancreas and soil. The environmental stress is confirmed by the expression of the relative biomarkers of exposure MT1 and HSP70. Our study, with a multimarker approach, consents getting a complete analysis of environmental quality and the effect of pollution on organisms and shows for the first time the heavy metals contamination status in certain Greece area. Furthermore, P. pruinosus proved to be an appropriate indicator of pollution in terrestrial ecosystems.     

南美白对虾肝胰腺蛋白提取方法优化及无水保活环境胁迫对细胞凋亡蛋白的影响

为提高Western Blotting结果的可靠性,以蛋白溶出率和凝胶电泳蛋白条带完整性为指标,比较提取液种类、研磨方式、酶抑制剂种类及其体积分数和提取时间对南美白对虾肝胰腺蛋白提取效果的影响。采用优化后的提取方法获得高质量蛋白样品,并采用Western Blotting法分析无水环境胁迫后南美白对虾肝胰腺组织中细胞凋亡信号通路相关蛋白的表达水平。结果表明：RIPA裂解液作为提取溶剂所得的蛋白溶出率高于水提和磷酸盐缓冲液,电动匀浆和液氮研磨所得蛋白条带更完整,4%蛋白酶磷酸酶混合抑制剂能有效抑制肝胰腺内源酶引起的蛋白降解;采用Western Blotting法分析无水保活期间南美白对虾肝胰腺蛋白,发现低温诱导休眠的同时会引起细胞轻微凋亡,且凋亡水平呈应激时间依赖性增加,环境胁迫解除后有所回调。     

不同产区青蟹肝胰腺风味品质的比较

采用感官评价、味觉活度值、电子鼻和挥发性物质分析,对来自渤海、东海和南海地区养殖的青蟹肝胰腺进行了风味品质比较。结果表明,东海雄蟹和雌蟹肝胰腺的感官评价较好;通过游离氨基酸的TAV分析发现,肝胰腺鲜甜味均较好,Glu、Ala和Arg是提供愉快滋味的主要游离氨基酸,渤海和东海的雌雄蟹肝胰腺中鲜甜味氨基酸含量相近,且均高于南海雌雄蟹;利用主成分分析发现,三个地区的雌雄蟹肝胰腺的第一和第二主成分之和均大于90%,其气味之间存在较大差异;通过挥发性成分分析发现,3个地区雄蟹肝胰腺中分别检测到41、45、46种挥发性风味物质,雌蟹肝胰腺中分别检测到45、40、43种挥发性风味化合物。醛类物质是青蟹肝胰腺主要的风味来源,其中1-癸烯-3-酮、2-戊基呋喃使东海青蟹肝胰腺呈现独特的风味。综上认为,东海雌雄蟹肝胰腺的风味品质相对较好。     

鱿鱼肝脏自水解过程中内源蛋白酶的作用研究

对鱿鱼肝脏蛋白酶的酶学性质及其在鱿鱼肝脏自水解过程中的作用进行研究。结果表明：以偶氮酪蛋白为底物,TCA可溶性肽为评价指标,测得鱿鱼肝脏蛋白酶粗酶的最适反应温度为40℃,最适反应pH值为5.0;该粗酶在pH3.0～8.0较为稳定。在鱿鱼肝脏自水解过程中,内源性半胱氨酸蛋白酶发挥主要作用。此外,丝氨酸蛋白酶和金属蛋白酶也有一定作用。     

Aminopeptidase from jumbo squid (Dosidicus gigas) hepatopancreas: purification,characterisation, and casein hydrolysis

An aminopeptidase (AP) was partially purified from jumbo squid (Dosidicus gigas) hepatopancreas with 154.24‐fold and yield of 6.15%. The purification procedure consisted of ammonium sulphate fractionation and DEAE‐Sephacel chromatography. The enzyme was approximately 48–53 kDa as estimated by SDS‐PAGE. With l ‐leu‐p‐NA, it had optimum activity at pH 8.0 and 30 °C. The K_m and V_max/K_m values of the enzymes for l ‐leu‐p‐NA were 0.326 mm and 2787 at 37 °C, respectively. Activation energy (E_a) of the enzyme was 53.50 kJ M^?1.The AP showed activity against seven synthetic substrates: l ‐proline>l ‐methionine>Ac. l ‐γ‐glutamic>l ‐glycine>l ‐leucine>l ‐alanine>l ‐lysine‐p‐NA. The enzyme was strongly inhibited by Bestatin, partially inhibited by a metal‐chelating agent and by PCMB, a cystein protease inhibitor. Zn²⁺ and (or) Ca²⁺ seemed to be its metal cofactor(s). Incubation of casein with the partially purified AP resulted in a degree of hydrolysis of 6%.