Anisakis pegrefii and Anisakis simplex sensu strictu in Mediterranean sea

BACKGROUND: Anisakiasis is a parasitic zoonosis caused by ingestion of nematode larvae belonging to Anisakidae family, following consumption of raw, undercooked or improperly processed fish. Mediterranean sea represent an aquatic ecosystem particularly suitable for the development of Anisakid larvae. The aim of this work is to found intra-species and inter-species nucleotide differences by phylogenetic analysis in this geographical area. METHODS: In the period from January to November 2013, 584 fish from Mediterranean sea were screened, and they were found parasitised of 6318 type I anisakid larvae. Fish were eviscerated and observed by stereo microscope to collect larvae relived in the viscera, organs and muscles. After genus identification, the larvae were subjected to molecular analysis by extraction of DNA, amplification of ITS gene and restriction enzyme. PCR products were sequenced and the sequences were analyzed and aligned to examine the relationship of nucleotides. RESULTS: The anisakid species we have identified were Anisakis pegreffii and Anisakis simplex s.s.. Phylogenetic analysis detects nucleotide differences between the two species. In the positions 251 and 267 was found a Cytosine in Anisakis pegreffii and a Thymine in Anisakis simplex s.s., respectively. No difference was found in Anisakis pegreffii specie becoming from different fish of different areas. CONCLUSIONS:. The parasite DNA were amplified and sequenced to identify any nucleotide differences between the different species as well as within the same species.No intra-species sequence differences were found in Anisakis pegreffii. Two inter-species differences were found between Anisakis pegreffii and Anisakis simplex. Further studies will be conducted to confirm nucleotide differences in other target genes

Exercise-induced Modifications on Haematochemical and Electrophoretic Parameters During 1600 and 2000 Meters Trot Races in Standardbred Horses

Abstract Piccione, G., Casella, S., Giannetto, C., Monteverde, V. and Ferrantelli, V. 2009. Exercise-induced modifications on haematochemical and electrophoretic parameters during 1600 and 2000 meters trot races in Standardbred horses. J. Appl. Anim. Res., 35: 131–135. The aim of the present study was to evaluate the modifications of some haematochemical and electrophoretic parameters during two different workloads in Standardbred horses. Ten clinically healthy horses were divided into two equal groups, taking part in an official trot race at 1600 or 2000 meters course, respectively. Blood samples were collected from each horse at rest, after warm up, 0, 30 and 60 min after the end of the race. Serum concentrations of albumin, globulins, total proteins, total cholesterol, triglycerides, glucose, hydroxybutyrate, urea, creatinine, creatine kinase (CK), glutamate pyruvate transaminases (GPT) and glutamate oxalacetate transaminases (GOT) were assessed. Exercise increased triglyceride, glucose, creatinine and..

Human Drug Pollution in the Aquatic System: The Biochemical Responses of Danio rerio Adults

To date, drug pollution in aquatic systems is an urgent issue, and Danio rerio is a model organism to study the toxicological effects of environmental pollutants. The scientific literature has analyzed the effect of human drug pollution on the biochemical responses in the tissues of D. rerio adults. However, the information is still scarce and conflicting, making it difficult to understand its real impact. The scientific studies are not consistent with each other and, until now, no one has grouped their results to create a baseline of knowledge of the possible impacts. In this review, the analysis of literature data highlights that the effects of drugs on adult zebrafishes depend on various factors, such as the tissue analyzed, the drug concentration and the sex of the individuals. Furthermore, the most influenced biochemical responses concern enzymes (e.g., antioxidants and hydrolase enzymes) and total protein and hormonal levels. Pinpointing the situation to date would improve the understanding of the chronic effects of human drug pollution, helping both to reduce it in the aquatic systems and then to draw up regulations to control this type of pollution

Aroclor 1254 inhibits the chemiluminescence response of peritoneal cavity cells from sharpsnout sea bream (Diplodus puntazzo)

Chronic exposure to polychlorinated biphenyls (PCBs) affect the immune system of fish and could lead to a decreased disease resistance. The effects of Aroclor 1254, PCB mixtures, on the Diplodus puntazzo innate immunity were examined by assaying the zymosan stimulated chemiluminescence response (CL) of peritoneal cavity cells (PCCs) at various times (1, 24, 48 h and 1-4 weeks) from intraperitoneal injection of the xenobiotic (1 mg kg(-1) body weight). Controls were performed by assaying cells from medium-treated fish. Since the kinetic of the chemiluminescence response showed the highest peak at 25 min after the zymosan stimulation of the cells, the values found at that time were considered. The CL enhancement observed at 1 h after the treatment with xenobiotic was followed by a decreased response at 24 h and appeared to be lower at 1-4 weeks when compared to the CL response of the control, suggesting a protracted effect of PCBs on the peritoneal cavity. Since PCCs incubated in vitro for 1 h with 0.05 and 0.1 mu g ml(-1) Aroclor showed an enhanced CL, the effect of the xenobiotic could be exerted on the cell responsiveness to zymosan. It is known that fish CL response of PCCs can be imputed to phagocyte (macrophages and neutrophils) activation, these cells and their responsiveness to zymosan can be used in immunotoxicology assay to monitor the fish health in polluted environment. (C) 2014 Elsevier Ltd. All rights reserved

In vitro effect of cadmium and copper on separated blood leukocytes of Dicentrarchus labrax.

The immunotoxic effects of heavy metals on blood leukocytes of sea bass (Dicentrarchus labrax) were examined. The cells, separated by a discontinuous Percoll-gradients, were exposed in vitro to various sublethal concentrations of cadmium and copper (10(-7) M, 10(-5) M, and 10(-3) M) and their immunotoxic effect was then evaluated by measuring neutral red uptake, MU assay, DNA fragmentation and Hsp70 gene expression. First of all, we demonstrated that the cells treated in vitro could incorporate Cd and Cu. A relationship between heavy metal exposure and dose-time-dependent alterations in responses of leukocytes from blood was found for both metals, but copper was more immunotoxic than cadmium in all assays performed. A significant reduction in the cells' ability to uptake neutral red and viability by MU assay was recorded, indicating that both cadmium and copper could change the membrane permeability, inducing cellular apoptosis when the concentration of metals reached 10(-3) M. The apoptotic effect may also explain the high level of cytotoxicity found when the leukocytes were exposed to higher concentration of metals. These results demonstrated that toxic effect of copper and cadmium affect on the mechanisms of cell-mediated immunity reducing the immune defences of the organism. (C) 2014 Elsevier Inc. All rights reserved

Analysis of Body Perception, Preworkout Meal Habits and Bone Resorption in Child Gymnasts

The beneficial effects of physical activity on body image perception and bone are debated among artistic gymnasts. Gymnasts seem to be at greater risk of developing body dissatisfaction, eating disorders and osteoporosis due to inadequate nutrition and attention to the appearance of the body. The objective of this work was to investigate the association between the artistic gymnast and a more favorable body image compared to their sedentary peers and if a preworkout high-carbohydrate meal (HCM; 300 kcal, 88% carbohydrates, 9% protein, 3% fat) or high-protein meal (HPM; 300 kcal, 55% carbohydrates, 31% protein, 13% fat) is able to attenuate bone resorption in young rhythmic gymnasts. Twenty-eight preadolescent female gymnasts were examined. Self-esteem tests were used to analyze body image perception. Preworkout eating habits were examined by short food frequency questions (FFQ) validated for children. The biomarker of the bone resorption C-terminal telopeptide region of collagen type 1 (CTX) was measured in the urine (fasting, postmeal and postworkout). Gymnasts reported higher satisfaction with their body appearance compared to sedentary peers. Of the gymnasts, 30% did not have a preworkout meal regularly, and the timing of the consumption was variable. Bone resorption was decreased by the HCM, consumed 90 min before the training, with respect to the HPM. The study suggests that playing artistic gymnastics is associated with a positive body self-perception in a child. The variability in preworkout meal frequency and timing need attention to prevent inadequate eating habits in light of the ability of the HCM to reduce acute bone resorption

A histological study of eosinophilic granuloma in mice following infestation with Anisakis larvae

Anisakidosis is a parasitic anthropozoonosis caused by the nematode larvae of the Anisakidae family. Anisakid nematodes belonging to the Anisakis pegreffii are highly prevalent in several fish species living in the Mediterranean sea. The larvae can accidentally infect humans following the ingestion of infected raw or undercooked sea fish. A migrating larva causes a clinical disease when it invades the stomach or intestinal wall and peritoneum, mimicking an eosinophilic gastroenteritis or an ulcer. In this preliminary experiment, the histopathology of the newly-formed parasitic granulomas in mice infested with third stage Anisakis pegreffii larvae, was studied and described. The larvae were morphologically identified as Anisakis type I by the presence of a boring tooth and a mucron, without ventriculus and caecum. The larval DNA was extracted and amplified by polymerase chain reaction (PCR). After PCR, the samples were processed to undergo restriction fragment length polymorphism analysis (RFLP). This was done to scan the restriction profiles for genetic identification. PCR products were purified and sequenced. The sequences were analysed to detect the relationship between nucleotides and perform a phylogenetic analysis. The paraffin-embedded granuloma samples showed worms having a diameter of 0.55 mm x 0.37 mm, polymyarian muscle cells and a circular intestine. The histological profiles showed a primary lesion at the site of anisakid penetration marked by oedema and neutrophil and eosinophil infiltration. The presence of histiocytes or epithelioid histiocytes, lymphocytes, monocytes, and plasma cells was also possible. Fibrinous exudation, hemorrhage, or vascular damage were detected within the first week of the acute intestine infection with a massive eosinophilic infiltration. Two weeks after the infestation, the infiltrating host cells formed a granuloma in the tissue surrounding the penetrated worm mainly consisting of eosinophils, a large number of fibroblasts and a varying number of admixed multinucleated giant cells. In order to explain the origin of the eosinophilic granulomas, a study into the produced substance attracting eosinophils and other host white blood cells to the area will carry out

Salmo salar fish waste oil: Fatty acids composition and antibacterial activity

Background and aims Fish by-products are generally used to produce fishmeal or fertilizers, with fish oil as a by-product. Despite their importance, fish wastes are still poorly explored and characterized and more studies are needed to reveal their potentiality. The goal of the present study was to qualitatively characterize and investigate the antimicrobial effects of the fish oil extracted from Salmo salar waste samples and to evaluate the potential use of these compounds for treating pathogen infections. Methods Salmo salar waste samples were divided in two groups: heads and soft tissues. Fatty acids composition, and in particular the content in saturated (SAFAs), mono-unsaturated (MUFAs) and Polyunsaturated (PUFAs) fatty acids, was characterized through GC/MS Thermo Focus GC-DSQ II equipped with a ZB-5 fused silica capillary tubes column. The antimicrobial activity of the salmon waste oils was evaluated through the Minimum Inhibitory Concentration assay and the antibiotics contamination was determined by Liquid Chromatography with tandem Mass Spectrometry (LC-MS/MS) analysis. All experiments were done at least in triplicate. Results GC/MS analysis has shown the specific fatty acid composition of the salmon waste oils and their enrichment in MUFAs and PUFAs, with special reference to omega-3, -6, -7, -9 fatty acids. Furthermore, our study has highlighted the antimicrobial activity of the fish waste oil samples against two Gram+ and Gram- bacterial strains. Conclusions These data confirm that the fish waste is still quantitatively and qualitatively an important source of available biological properties that could be extracted and utilized representing an important strategy to counteract infective diseases in the context of the circular economy

Determination of Ten Corticosteroids in Illegal Cosmetic Products by a Simple, Rapid, and High-Performance LC-MS/MS Method

The aim of our present work was the development of a rapid high-performance liquid chromatography method with electrospray ionization and tandem mass spectrometry detection (LC-ESI-MS/MS) for the determination of several corticosteroids in cosmetic products. Corticosteroids are suspected to be illegally added in cosmetic preparations in order to enhance the curative effect against some skin diseases. Sample preparation step consists in a single extraction with acetonitrile followed by centrifugation and filtration. The compounds were separated by reversed-phase chromatography with water and acetonitrile (both with 0.1% formic acid) gradient elution and detected by ESI-MS positive and negative ionization mode. The method was validated at the validation level of 0.1 mg kg−1. Linearity was studied in the 5–250 μg L−1 range and linear coefficients (r2) were all over 0.99. The accuracy and precision of the method were satisfactory. The LOD ranged from 0.085 to 0.109 mg kg−1 and the LOQ from 0.102 to 0.121 mg kg−1. Mean recoveries for all the analytes were within the range 91.9–99.2%. The developed method is sensitive and useful for detection, quantification, and confirmation of these corticosteroids in cosmetic preparations and can be applied in the analysis of the suspected samples under investigation

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