Making good use of the byproducts of cultivation: green synthesis and antibacterial effects of silver nanoparticles using the leaf extract of blueberry.

In this study, we made an effort to use blueberry leaves extract as a reducing and a capping agent to biosynthesize silver nanoparticles (AgNPs). The successful formation of AgNPs was confirmed with UV-visible spectroscopy, transmission electron microscopy, Fourier transform infrared spectroscopy and X-ray diffraction. Furthermore, their antimicrobial activities against four kinds of multi drug resistant clinical pathogens and aquatic pathogens were investigated. The results showed that as-prepared AgNPs exhibited broad-spectrum antibacterial activities. The antibacterial effects of the AgNPs based on the structural damage and DNA degradation of the tested bacteria were also studied. The green synthesis of AgNPs using blueberry leaf extract, which provided a new idea on the use of the byproducts of blueberry harvesting.

Toxicological proteomic responses of halophyte Suaeda salsa to lead and zinc.

The long term (30 days) toxicological effects of environmentally relevant concentrations of Pb2+ (20µg/L) and Zn2+ (100µg/L) were characterized in Suaeda salsa using proteomics techniques. The responsive proteins were related to metabolism (Krebs cycle and Calvin cycle), protein biosynthesis, stress and defense, energy, signaling pathway and photosynthesis in Pb2+, Zn2+ and Pb2++ Zn2+ exposed groups in S. salsa after exposures for 30 days. The proteomic profiles also showed differential responses in S. salsa to metal exposures. In Pb2+-treated group, the proteins were categorized into cystein metabolism and pentose phosphate pathway. The responsive proteins were basically involved in glutathione metabolism, glycolysis, cystein and methane metabolism, and voltage-dependent anion channel in Zn2+-treated group. In Pb2++ Zn2+-treated group, the proecular mechanism at protein level remtein responses were devided into tyrosine metabolism and glycolysis. Our results showed that the two typical heavy metals, lead and zinc, could induce toxicological effects in halophyte S. salsa at protein level.

Effects of potential probiotic Bacillus cereus EN25 on growth, immunity and disease resistance of juvenile sea cucumber Apostichopus japonicus.

This study was conducted to determine effects of potential probiotic Bacillus cereus EN25 (isolated from mud of sea cucumber culturing water bodies) on growth, immunity and disease resistance against Vibrio splendidus infection in juvenile sea cucumbers Apostichopus japonicus. Animals were respectively fed diets with B. cereus EN25 at 0 (control), 10(5), 10(7) and 10(9) CFU/g for 30 days. Results showed that dietary B. cereus EN25 had no significant effects on growth, total coelomocytes counts and acid phosphatase activity of A. japonicus (P > 0.05). Dietary EN25 at 10(7) CFU/g had significantly improved the phagocytosis, respiratory burst activity and total nitric oxide synthase activity of animals (P < 0.05). Compared to control, dietary EN25 at 10(5) or 10(7) CFU/g had no significant effects on superoxide dismutase activity of A. japonicus (P > 0.05), whereas dietary EN25 at 10(9) CFU/g had significantly decreased its activity (P < 0.05). The cumulative mortality after V. splendidus challenge decreased significantly in sea cucumbers fed with EN25 at 10(7) CFU/g (P < 0.05). The present study confirmed dietary B. cereus EN25 at 10(7) CFU/g could significantly improve immunity and disease resistance in juvenile A. japonicus.

The interactive effects of mercury and selenium on metabolic profiles, gene expression and antioxidant enzymes in halophyte Suaeda salsa.

Suaeda salsa is the pioneer halophyte in the Yellow River Delta and was consumed as a popular vegetable. Mercury has become a highly risky contaminant in the sediment of intertidal zones of the Yellow River Delta. In this work, we investigated the interactive effects of mercury and selenium in S. salsa on the basis of metabolic profiling, antioxidant enzyme activities and gene expression quantification. Our results showed that mercury exposure (20 µg L(-1)) inhibited plant growth of S. salsa and induced significant metabolic responses and altered expression levels of INPS, CMO, and MDH in S. salsa samples, together with the increased activities of antioxidant enzymes including SOD and POD. Overall, these results indicated osmotic and oxidative stresses, disturbed protein degradation and energy metabolism change in S. salsa after mercury exposures. Additionally, the addition of selenium could induce both antagonistic and synergistic effects including alleviating protein degradation and aggravating osmotic stress caused by mercury.

Gender-specific metabolic responses in hepatopancreas of mussel Mytilus galloprovincialis challenged by Vibrio harveyi.

Mussel Mytilus galloprovincialis is a marine aquaculture shellfish and frequently studied in shellfish immunology. In this work, the gender-specific metabolic responses induced by Vibrio harveyi in hepatopancreas from M. galloprovincialis were characterized using NMR-based metabolomics. In details, V. harveyi challenge increased the levels of amino acids including (valine, leucine, isoleucine, threonine, alanine, arginine and tyrosine) and ATP, and decreased the level of glucose in male mussel hepatopancreas. In V. harveyi-challenged female mussel hepatopancreas, both threonine and AMP were significantly elevated, and choline, phoshphocholine, sn-glycero-3-phosphocholine, taurine, betaine and ATP were depleted. Obviously, only threonine was similarly altered to that in V. harveyi-challenged male mussel hepatopancreas. These findings confirmed the gender-specific metabolic responses in mussels challenged by V. harveyi. Overall, V. harveyi induced an enhanced energy demand through activated glycolysis and immune response indicated by increased BCAAs in male mussel hepatopancreas. In female mussel hepatopancreas, V. harveyi basically caused disturbances in both osmotic regulation and energy metabolism through the metabolic pathways of conversions of phosphocholine and ADP to choline and ATP, and sn-glycero-3-phosphocholine and H2O into choline and sn-glycerol 3-phosphate. The altered mRNA expression levels of related genes (Cu/Zn-SOD, HSP90, lysozyme and defensin) suggested that V. harveyi induced obvious oxidative and immune stresses in both male and female mussel hepatopancreas. This work demonstrated that V. harveyi could induce gender-specific metabolic responses in mussel M. galloprovincialis hepatopancreas using NMR-based metabolomics.

A real-time PCR for the detection of infectious myonecrosis virus in penaeid shrimp.

Infectious myonecrosis virus (IMNV) is a recently observed shrimp virus, which threats the cultured Litopenaeus vannamei and can cause huge economic loss in shrimp farming industry. The specific aim of this study was to develop a new sensitive real-time PCR method for the specific detection of shrimp IMNV. A real-time PCR assay with a pair of primers to specifically amplify a 101bp IMNV cDNA fragment and a corresponding TaqMan probe was developed, which shown to be specific for IMNV without cross reaction with DNA samples prepared from four other shrimp viruses including white spot syndrome virus (WSSV), hepatopancreatic parvovirus (HPV), monodon baculovirus (MBV), and infectious hypodermal and haematopoietic virus (IHHNV). The method could detect as low as one single copy of IMNV plasmid cDNA.

Immunohistochemical observations of methionine-enkephalin and delta opioid receptor in the digestive system of Octopus ocellatus.

The study was designed to determine whether methionine-enkephalin (met-Enk) or delta opioid receptor was present in the digestive system of Octopus ocellatus. The results showed that they were both in the bulbus oris, esophagus, crop, stomach, gastric cecum, intestine, posterior salivary glands of O. ocellatus, one of them, met-Enk in the rectum, anterior salivary glands, digestive gland. And the distributions were extensive in the digestive system. Strong or general met-Enk immunoreactivity was observed in the inner epithelial cells of the bulbus oris, esophagus, stomach, gastric cecum, intestine, anterior salivary glands and the adventitia of the intestine and rectum, and so was the delta opioid receptor immunoreactivity in the inner epithelial cells of the bulbus oris, esophagus, and crop, however, they were weak in other parts. Combining with delta opioid receptor, met-Enk may be involved in the regulations of food intake, absorption, movement of gastrointestinal smooth muscle and secretion of digestive gland. The different densities of met-Enk and delta opioid receptor may be related to the different functions in the digestive system of O. ocellatus.

The dynamics of reproductive rate, offspring survivorship and growth in the lined seahorse, Hippocampus erectus Perry, 1810.

Seahorses are the vertebrate group with the embryonic development occurring within a special pouch in males. To understand the reproductive efficiency of the lined seahorse, Hippocampus erectus Perry, 1810 under controlled breeding experiments, we investigated the dynamics of reproductive rate, offspring survivorship and growth over births by the same male seahorses. The mean brood size of the 1-year old pairs in the 1(st) birth was 85.4±56.9 per brood, which was significantly smaller than that in the 6(th) birth (465.9±136.4 per brood) (P<0.001). The offspring survivorship and growth rate increased with the births. The fecundity was positively correlated with the length of brood pouches of males and trunk of females. The fecundity of 1-year old male and 2-year old female pairs was significantly higher than that from 1-year old couples (P<0.001). The brood size (552.7±150.4) of the males who mated with females that were isolated for the gamete-preparation, was larger than those (467.8±141.2) from the long-term pairs (P<0.05). Moreover, the offspring from the isolated females had higher survival and growth rates. Our results showed that the potential reproductive rate of seahorses H. erectus increased with the brood pouch development.

Application of taxonomic distinctness indices of littoral macroinvertebrate communities for assessing long-term variation in ecological quality status of intertidal ecosystems, northern China.

It has been increasingly recognized that taxonomic diversity indices have a number of desirable properties as an indicator for assessing ecological quality status, in particular their less sensitivity to natural habitat type and sampling effort but more to environmental stress and anthropogenic impact, and a statistical framework for the assessment of the significance of departure from expectation. Taxonomic patterns of macroinvertebrate fauna for assessing ecological quality status were studied based on six datasets collected from intertidal zones of the Yellow Sea, near Qingdao, northern China, during the period of 1989-1998. The invertebrate communities were sampled yearly at five stations with different bottom types during summer season (June). A total of 141 macroinvertebrate taxa were identified belonging 119 genera, 81 families, 34 orders, 19 classes, and 10 phyla. Multivariate analyses demonstrated that the taxonomic patterns of invertebrate fauna represented a significant variation in long-term temporal scale during the study period. The average taxonomic distinctness indices (Δ(+)) decreased to a significantly low level, while the variation in taxonomic distinctness measures (Λ(+)) increased to a significantly high level compared with the expected values from 1989 to 1998. The pairwise indices of Δ(+) and (Λ(+)) showed a decreasing and increasing trend of departure from the expected taxonomic breadth in response to the environmental stress and anthropogenic impact, respectively. These results imply that the ecological quality status has been significantly deteriorated due to the increasing environmental stress and anthropogenic impact in intertidal zones of the Yellow Sea, northern China, and that the taxonomic distinctness indices of macroinvertebrate fauna are a robust indicator for evaluating ecological quality status.

Immunohistological detection of mu, delta and kappa opioid-like receptors in the gill, gonad, and hemocytes of the scallop Chlamys farreri.

Endogenous opioid peptides and opioid receptors form a neuromodulatory system, which plays an important part in the control of physiological pathways. In addition, some opioid peptides can function as endogenous messengers of the immune system and participate in the regulation of the immune response. The present studies indicated that mu, delta, and kappa opioid-like receptors were present in the gill and gonad of the scallop Chlamys farreri. Furthermore, the significance of opioid peptides involvement with the immune system is ascertained from the presence of mu, delta, and kappa opioid-like receptors on hemocytes of the scallop. Our report constitutes the first characterization of mu, delta, and kappa opioid-like receptors in the gill and gonad of the scallop Chlamys farreri.

HB-1, an acute myeloid leukemic cell line with the capability of infiltrating into the brain in CBA/N mice.

An acute myeloid leukemic HB-1 cell line was cloned and established from the spleen cells of irradiated CBA/N mice. Acute myeloma leukemia-like syndrome would be induced in normal CBA/N mice after intravenous injection of HB-1 cells, and the death of mouse happened within about two weeks. In general, leukemic cells transplanted into the mice would infiltrate into the hematopoietic organs, lungs, kidneys and liver. An interesting observation in our study was that HB-1 cells were present not only in the lung, kidney, and liver but also in the cerebrum and cerebellum. It was beyond our expectation that the leukemic cells could go through the blood-brain barrier in most circumstances. On the basis of the observation, we expect that HB-1 cells could be used as a very useful model to elucidate the mechanism of infiltrating the blood-brain barrier for certain type of cells.