Therapeutic Intervention with Dietary Chitosan Nanoparticles Alleviates Fish Pathological and Molecular Systemic Inflammatory Responses against Infections

Marine bio-sourced chitosan nanoparticles (CSNP) are antimicrobial and immunomodulatory agents beneficial for fish medicine. Herein, dietary CSNP was investigated for the amelioration of the systemic inflammatory responses of an induced fish model. One hundred and forty-four rainbow trout were assigned to one pathogen-free and non-supplemented group (negative control), and three challenged groups: non-supplemented (positive control), CSNP-preventive, and CSNP-therapeutic. After a feeding experiment extended for 21 days, the organosomatic indices (OSI) and molecular aspects were assessed. After a challenge experiment extended for further 28 days, CSNP-therapeutic intervention was assessed on fish survival and systemic inflammatory responses on pathology, histo-morphology, and molecular aspects. With CSNP administration, OSI nonsignificantly decreased and the relative expression of targeted inflammatory-mediator genes was significantly increased. The CSNP-therapeutic fish showed an RPS of 80% as compared to the positive control group, and CSNP-therapeutic administration retained the highest gene expression augmentation up to 28 days after the challenge. Notably, the splenic reticulin fibers framework of the CSNP-therapeutic group retained the highest integrity among the groups during the infection. After recovery, reticulin fibers density in the CSNP-therapeutic samples was significantly higher than in the negative control group, which indicates high innate immunity. Thus, CSNP showed promising biotherapeutic features enhancing fish resistance against infections.     

Marine-Derived Chitosan Nanoparticles Improved the Intestinal Histo-Morphometrical Features in Association with the Health and Immune Response of Grey Mullet (Liza ramada)

Marine-derived substances are known for their beneficial influences on aquatic animals’ performances and are recommended to improve intestinal health, immunity, and anti-oxidative status. The present study investigates the role of chitosan nanoparticles on the intestinal histo-morphometrical features in association with the health and immune response of Grey Mullet (Liza ramada). Chitosan nanoparticles are included in the diets at 0, 0.5, 1, and 2 g/kg and introduced to fish in a successive feeding trial for eight weeks. The final body weight (FBW), weight gain (WG), and specific growth rate (SGR) parameters are significantly increased while feed conversion ratio (FCR) decreases by chitosan nanoparticles compared to the control (p < 0.05). The morphometric analysis of the intestines reveals a significant improvement in villus height, villus width, and the number of goblet cells in chitosan-treated groups in a dose-dependent manner. Additionally, there is a positive correlation between the thickness of the enterocyte brush border and the chitosan dose, referring to an increasing absorptive activity. Histologically, the intestinal wall of Grey Mullet consists of four layers; mucosa, sub-mucosa, tunica muscularis (muscular layers), and serosa. The histological examination of the L. ramada intestine shows a normal histo-morphology. The epithelial layer of intestinal mucosa is thrown into elongated finger-like projections, the intestinal villi. The values of hemoglobin, hematocrit, red blood cells (RBCs), total protein (TP), albumin, and globulin are significantly increased in fish fed 1, and 2 g/kg of chitosan nanoparticles compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest levels of TP and albumin are observed in fish fed 1 g/kg diet (p < 0.05). The lysozyme activity and phagocytic index are significantly enhanced by feeding chitosan nanoparticles at 0.5, 1, and 2 g/kg, whereas the phagocytic activity is improved in fish fed 1 and 2 g/kg (p < 0.05). The highest lysozyme activity and phagocytic index are observed in fish fed 1 g/kg. SOD is significantly activated by feeding chitosan nanoparticles at 1 g/kg. Simultaneously, glutathione peroxidase (GPx) and catalase (CAT) activities also are enhanced by feeding chitosan at 1 and 2 g/kg, compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest GPx and CAT activities are observed in fish fed 1 g/kg (p < 0.05). Conversely, the malondialdehyde (MDA) levels are decreased by feeding chitosan at 1 and 2 g/kg, with the lowest being in fish fed 1 g/kg (p < 0.05). To summarize, the results elucidate that L. ramada fed dietary chitosan nanoparticles have a marked growth rate, immune response, and anti-oxidative response. These improvements are attributed to the potential role of chitosan nanoparticles in enhancing intestinal histo-morphometry and intestinal health. These results soundly support the possibility of using chitosan nanoparticles at 1–2 g/kg as a feasible functional supplement for aquatic animals.     

A Burkholderia pseudomallei toxin inhibits helicase activity of translation factor eIF4A

The structure of BPSL1549, a protein of unknown function from Burkholderia pseudomallei, reveals a similarity to Escherichia coli cytotoxic necrotizing factor 1. We found that BPSL1549 acted as a potent cytotoxin against eukaryotic cells and was lethal when administered to mice. Expression levels of bpsl1549 correlate with conditions expected to promote or suppress pathogenicity. BPSL1549 promotes deamidation of glutamine-339 of the translation initiation factor eIF4A, abolishing its helicase activity and inhibiting translation. We propose to name BPSL1549 Burkholderia lethal factor 1.     

Fasciation in <Emphasis Type="Italic">Crassula argentea</Emphasis>: molecular identification of phytoplasmas and associated antioxidative capacity

The present study reports on phytoplasma induced fasciation in Crassula argintea (Crassulaceae). DNA was extracted from symptomless and fasciated tissues and amplified by nested PCR using universal primers P1/P7 followed by R16F2n/R16R2 produced amplicons of 1.2 Kb. The nucleotide sequence analyses of the amplicons indicated that fasciated plants were infected by phytoplasma. Phylogenetic analysis placed the Crassula fasciation phytoplasmas in 16SrII-D group. Histochemical staining for reactive oxygen species indicated that phytoplasma infected (PI) tissues possess significantly higher levels of hydrogen peroxide (H₂O₂) rather than superoxide (O₂ ^·-) as compared with symptomless tissues. PI tissues were also associated with a significant increase in antioxidant enzyme activities (catalase, peroxidase, polyphenol oxidase, and glutathione reductase) and electrolyte leakage as compared with symptomless tissues.     

Use of serology and real time PCR to control an outbreak of bovine brucellosis at a dairy cattle farm in the Nile Delta region,Egypt

Background

Bovine brucellosis remains one of the most prevalent zoonotic infections affecting dairy cattle in developing countries where the applied control programs often fail. We analyzed the epidemiologic pattern of bovine brucellosis in a dairy cattle herd that showed several cases of abortions after regular vaccination with RB51 (B. abortus vaccine). In 2013 thirty dairy cows, from a Holstein-Friesian dairy herd with a population of 600 cattle, aborted five months post vaccination by a regular RB51 vaccine. Blood samples were drawn from milking cows and growing heifers, as well as heifers and cows pregnant up to 6 months. These samples were collected in June 2013 (n?=?257) and May 2014 (n?=?263) and were tested by real time (rt)-PCR as well as serological tests, in particular Rose Bengal Test (RBT), Enzyme-Linked Immunosorbent Assays (ELISA) and Fluorescence Polarization Assay. Tissue specimens were also collected from an aborted fetus and cultured. Isolates were subjected to bacteriological typing tests at the genus and species levels.

Results

Five months post vaccination with RB51 vaccine, Brucella (B.) DNA was detected in blood samples of cows by rt-PCR. The serological tests also revealed the spread of Brucella field strains within the herd in 2013. Four Brucella isolates were recovered from specimens collected from the aborted fetus. These isolates were typed as follows: one B. abortus RB51 vaccine strain and three isolates of B. abortus field strain. The seropositive cows with positive rt-PCR might indicate an infection by the Brucella field strain; while the positive rt-PCR results from seronegative animals may either be due to circulating RB51 vaccine DNA in vaccinated animals or to circulating field strain in infected animals before seroconversion.

Conclusion

The results herein suggest that PCR can be a good supplementary tool in an outbreak situation, if an assay is available that can differentiate vaccine and field strains with a high analytical sensitivity. We recommend using RBT and ELISA in parallel in outbreak situations, to identify as many infected animals as possible during the initial screenings. This test procedure should be repeated for at least three successive negative tests, with one month interval.

     

Changes in growth and nitrogen assimilation in barley seedlings under cadmium stress

Barley (Hordeum vulgare L. cv. Martin) plants grown in solution culture, were exposed to increasing cadmium (Cd) concentration (0, 5, 10, 25, 50, and 100 μM) for a duration of 12 days. The sequence of important biochemical steps of nitrate (NO₃) assimilation were studied in roots and shoots as a function of external Cd concentration. Cadmium uptake in roots and shoots increased gradually with Cd concentration in the medium. This Cd accumulation lowered substantially root and shoot biomass. The nitrate reductase (NR, EC 1.6.6.1) and nitrite reductase (NiR, EC 1.6.6.4) activities declined under Cd stress. Concurrently, tissue NO₃ contents and xylem sap NO₃ concentration were also decreased in Cd‐treated plants. These results suggest that Cd could exert an inhibitory effect on the assimilatory NO₃ reducing system (NR and NiR) through a restriction of NO₃ availability in the tissues. We therefore examined, in short‐term experiments (12 h), the impact of Cd on NO₃ uptake and the two reductases in nitrogen (N)‐starved plants that were pretreated or not with Cd. It was found that Cd induced inhibition of both NO₃ uptake and activities of NR and NiR, during NO₃ induction period. The possible mechanisms of Cd action on NO₃ uptake are proposed. Further, in Cd‐grown plants, the glutamine synthetase (GS, EC 6.3.1.2) showed a decreasing activity both in shoots and roots. However, increasing external Cd concentration resulted in a marked enhancement of glutamate dehydrogenase (NADH‐GDH, EC 1.4.1.2) activity, coupled with elevated levels of ammonium (NH₄ in tissues. On the other hand, the total protein content in Cd‐treated plants declined with a progressive and substantial increase of protease activity in the tissues. These findings indicate that under Cd stress the usual pathway of NH₄ assimilation (glutamine synthetase/glutamate synthase) can switch to an alternative one (glutamate dehydrogenase). The changes in all parameters investigated were concentration‐dependent and more marked in roots than shoots. The regulation of N absorption and assimilation by Cd in relation to growth and adaptation to stress conditions are discussed.     

A Multivariate Data Analysis of the Clam Ruditapes Decussatus as Sentinel Organism of the Bizerta Lagoon (Tunisia)

Water, Air, & Soil Pollution - From May 1998 to May 1999, the clams Ruditapes decussatus were collected each month at three stations of the Bizerta lagoon. This paper represents a synthesis...     

Effects of Cumin and Ginger as Antioxidants on Dough Mixing Properties and Cookie Quality

The effects of cumin and ginger as antioxidants on dough mixing properties and cookie quality were evaluated. Antioxidant activities in cookies were estimated by total phenolic compounds (TPC) contents and free radical scavenging activities. The cookie dough development was evaluated using Mixolab equipment which showed that addition of cumin did not change dough stability and C2, but decreased C3 and C4. While the addition of 5% ginger decreased dough stability (from 8.4 in the control sample to 6.7 min with 5% ginger addition), C2 (from 0.49 in the control sample to 0.31 N·m with 5% ginger addition), C3, and C4. Cookies formulated with addition of cumin and ginger had increased spread ratios, were softer, and had lower L* and b* values (were darker) than the control. Sensory analysis showed that cookies with cumin and ginger additions had overall acceptability similar to that of the the control with a slightly darker appearance, as confirmed by color determination. Using cumin and ginger significantly increased TPC contents from 78.5 in the control to 93.0 and 109.8 mg of gallic acid equivalent/100 g, respectively. Similar results were observed in the antioxidant activity measured by 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH), which increased from 41.0% in the control to 51.5% and 64.6%, respectively, for cookies with 5% addition of cumin and ginger.