Combined effects of exposure to sub-lethal concentration of the insecticide chlorpyrifos and the herbicide glyphosate on the biochemical changes in the freshwater crayfish Pontastacus leptodactylus.

Glyphosate is an herbicide that inhibits the growth of weed plants, while chlorpyrifos is an insecticide commonly applied to control the pests' population. This study aimed to investigate the combined effects of chlorpyrifos and glyphosate on biochemical, immunological parameters, and oxidative stress biomarkers in freshwater crayfish Pontastacus leptodactylus for 21 days. The experimental design of this study was factorial (3 × 3), including 0.0, 0.4, and 0.8 mg L-1 glyphosate and 0.0, 2.5, and 5 µg L-1 chlorpyrifos. The exposure to chlorpyrifos, glyphosate alone and a mixture of them significantly decreased acetylcholinesterase, alkaline phosphatase, phenoloxidase activities, and total protein levels. The lactate dehydrogenase, glutamic-pyruvic-transaminase, and catalase activities, the contents of glucose, and malondialdehyde levels were increased in the crayfish. No significant changes were detected in glutamic-oxaloacetic-transaminase (SGOT) activity, triglyceride, and total antioxidant (TAO) levels in the crayfish treated with 0.4 mg L-1 glyphosate and the control group. Co-exposure of crayfish to chlorpyrifos and glyphosate increased SGOT activity and TAO levels. Although chlorpyrifos combined with glyphosate decreased the γ-Glutamyltransferase (GGT) activity, the GGT activity was significantly increased in the P. leptodactylus exposed during 21 days to 5 µg L-1 chlorpyrifos alone and 0.8 mg L-1 glyphosate alone. In comparison with the reference group, no significant changes were evidenced in the cholesterol levels in the P. leptodactylus treated with 2.5 µg L-1 chlorpyrifos, but its levels were significantly increased in the other treatment groups. In conclusion, the mix of glyphosate and chlorpyrifos exhibited synergic effects on the different toxicological biomarkers in the narrow-clawed crayfish. Co-exposure to pesticides may result in disruption of homeostasis in the crayfish by altering the biochemical and immunological parameters.

Evaluation of single and combined effects of cadmium and micro-plastic particles on biochemical and immunological parameters of common carp (Cyprinus carpio).

The growing accumulation of microplastics (MPs) in aquatic environments is a global concern. MPs are capable to interact with other environmental contaminants, including heavy metals, altering their toxicity. The aim of the study was to investigate the sub-lethal effects of cadmium chloride (Cd) alone and in combination with MPs on common carp (Cyprinus carpio). Multi-biomarkers, including plasma biochemical parameters and intrinsic immunological factors, were measured after 30 days of exposure. Exposure to Cd or NPs reduced the plasma activities of acetylcholinesterase (AChE) and gamma-glutamyl-transferase (GGT) and increased aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and alkaline phosphatase (ALP). Exposure to both compounds enhanced the observed effects except for AST activity and ALP at the highest concentrations, whereas evidenced an antagonistic interaction in ALT. Plasma total protein, albumin, and globulin levels were decreased, and the levels of glucose, triglyceride, and cholesterol levels increased mainly in the Cd groups with no additional effects derived from the co-exposure to both stressors. Lysozyme and alternative complement (ACH50) activities and the levels of total immunoglobulins, and complement C3 and C4 in fish exposed to Cd and MPs were lower than those in the control group and this decrease was more significant by the mixture of both compounds. These findings showed that the exposure to Cd or MPs alone is toxic to fish altering the biochemical and immunological parameters. Moreover, these alterations are even greater when the Cd and the MPS are combined suggesting synergistic effects in increasing Cd toxicity and vice versa.

Acute exposure to chlorpyrifos and glyphosate induces changes in hemolymph biochemical parameters in the crayfish, Astacus leptodactylus (Eschscholtz, 1823).

Acute and subacute toxicity tests for chlorpyrifos and glyphosate were performed on the crayfish, Astacus leptodactylus. The crayfish were divided into a control group and four experimental groups with exposure from 50.18 to 301.11 µg L-1 of chlorpyrifos for 96 h under semi-static test conditions. In the same experiment, the crayfish were exposed to different concentrations of glyphosate (from 0.0, 8.14 to 13.05 mg L-1) for 96 h. Mortality was recorded, and the median lethal concentrations (LC50) were calculated using probit analysis. The 96 h LC50 values of chlorpyrifos and glyphosate to A. leptodactylus were 49.55 ±â€¯4.66 µg L-1 and 7.83 ±â€¯0.50 mg L-1, respectively. The results showed that chlorpyrifos was a few hundred times more toxic than glyphosate for the crayfish. Then, the crayfish were exposed to different subacute concentrations of chlorpyrifos (12.5 and 25 µg L-1) and glyphosate (2 mg L-1 and 4 mg L-1) for seven days, and their toxicity was evaluated through several hemolymph biochemical parameters. Chlorpyrifos and glyphosate exposure caused a significant (P < 0.01) increase in the activity of lactate dehydrogenase (LDH), and gamma-glutamyl transferase (GGT). The results showed a significant (P < 0.01) decrease in acetylcholinesterase (AChE), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and phenoloxidase (PO) activities in the hemolymph of the crayfish exposed to chlorpyrifos and glyphosate. The total protein and cholesterol levels showed a significant decrease (P < 0.01) in the hemolymph of the crayfish after seven days of exposure to chlorpyrifos and glyphosate. There was a significant increase in triglycerides and glucose levels in treated crayfish. These results show that chlorpyrifos is highly toxic, while glyphosate is moderately toxic to A. leptodactylus. According to the results, subacute concentrations of chlorpyrifos and glyphosate could lead to a collapse of cellular homeostasis and changes in the hemolymph biochemical parameters in the crayfish. Physiological changes caused by exposure to chlorpyrifos and glyphosate in A. leptodactylus may have direct effects on the survival of these organisms.

Comparative single-dose pharmacokinetics of orally administered florfenicol in rainbow trout (Oncorhynchus mykiss, Walbaum, 1792) at health and experimental infection with Streptococcus iniae or Lactococcus garvieae.

This study evaluates changes in the pharmacokinetic behavior of a single oral dose of florfenicol in rainbow trouts experimentally infected with Lactococcus garvieae or Streptococcus iniae. One hundred and fifty fish were randomly divided into three equal groups: 1-healthy fish, 2-fish inoculated with S. iniae (2.87 × 107 CFU/ml, i.p.), and 3-fish inoculated with L. garvieae (6.8 × 105 CFU/ml, i.p.). Florfenicol was administered to all groups at 15 mg/kg by oral gavage. Blood sampling was performed at 0, 2, 3, 6, 8, 12, 24, 48, 72, and 120 hr after drug administration to each group, and plasma concentration of florfenicol was assayed by HPLC method. The MICs of florfenicol were 1.2 µg/ml and 5 µg/ml against L. garviae and S. iniae, respectively. Healthy fish showed higher values for most of the PK/PD parameters as compared to fish infected with L. garvieae which was reversed in fish infected with S. iniae. Fish infected with L. garvieae showed decreased relative bioavailability accompanied by increased volume of distribution at steady-state (Vdss ) and total body clearance (ClB ). Infection with S. iniae increased the peak concentration of drug after administration (Cmax) and decreased elimination half-life (T1/2 ß ), central compartment volume (Vc ), and Vdss . In conclusion, infection with these bacteria can affect the pharmacokinetic behavior of florfenicol in rainbow trouts as shown by decreased bioavailability and increased total body clearance and volume of distribution in L. garvieae infection and decreased volume of distribution accompanied by increased Cmax in S. iniae-infected fish.

First isolation and identification of Aeromonas veronii and Chryseobacterium joostei from reared sturgeons in Fars province, Iran.

The purpose of the present study was to isolate and identify the pathogenic agents in Acipenser stellatus (Pallas, 1771) and Huso huso, (Linnaeus, 1758) reared in the south of Fars province, Iran which have shown infectious disease signs. Samples from spleen and kidney of 32 fishes showing septicemia symptoms such as decreasing of appetite, unbalanced swimming, expanded wounds, and petechia on the body surfaces, pectoral fins rot, visceral hemorrhage, bleeding on the spleen, and heart ascites were collected. Then samples were cultured on brain heart infusion agar growth media, stain and biological and biochemical tests on purified bacteria were performed. On the other hand, 16S rDNA region of the isolated organism was amplified using PCR. The amplified gene fragment was sequenced and evolutionary history was inferred by phylogenetic tree construction using neighbor-joining method. Results indicated that two bacterial species including Chryseobacteriumjoostei which isolated from the kidney of stellate sturgeon (43.00%), and Aeromonasveronii which isolated from the spleen of both sturgeon species (75.00% and 31.00% from beluga and stellate sturgeon, respectively), were recognized. Phylogenetic tree analysis showed that Fars isolated organisms including A. veronii and C. joostei had highest similarity with A. veronii bv veronii and C. joostei isolated from France, respectively.

The effects of single or combined administration of galactooligosaccharide and Pediococcus acidilactici on cutaneous mucus immune parameters, humoral immune responses and immune related genes expression in common carp (Cyprinus carpio) fingerlings.

The present study was performed to investigate the effects of single or combined administration of galactooligosaccharide (GOS) and Pediococcus acidilactici on cutaneous mucus immune parameters, humoral immune responses and immune related genes expression in common carp (Cyprinus carpio) fingerlings. Carps were fed experimental diets for 8 weeks as follows: non-supplemented (Control), prebiotic diet (10 g/kg GOS), probiotic diet (1 g/kg [0.9 × 107 CFU] lyophilized P. acidilactici) and synbiotic diet (10 GOS in combination with 1 g/kg [0.9 × 107 CFU] lyophilized P. acidilactici). Unlike skin mucus, the serum lysozyme activity showed no significant difference between carps fed supplemented or control diets, however, remarkable elevation of serum ACH50 activity was noticed in carps fed supplemented diet (pro-, pre- and synbiotic diets) compared control group. Besides, feeding on pro-, pre- and synbiotic supplemented diets significantly increased serum and skin mucus total Ig levels. However, no significant difference was observed between treatments and control group in case of skin mucus proteases activity. There was no significant difference between expression levels of intestinal genes of LYZ and IL1b in fish fed on pre- and synbiotic, compared to the control. However, evaluation of TNF-alpha gene expression in the intestine of carps revealed remarkable down-regulation in treated groups (p < 0.05). These results indicated positive effect of supplementation of carp diet with GOS and P. acidilactici on some mucosal or serum immune parameters.

Detection of tetracycline resistance genes in bacteria isolated from fish farms using polymerase chain reaction.

Five common tetracycline resistance genes tet(A), tet(B), tet(M), tet(O) and tet(S) were studied by polymerase chain reaction in 100 bacteria isolated from Iranian fish farms. In the antibiogram test most of the bacteria were either intermediately or completely resistant to tetracycline. Nine isolates out of 46 Aeromonas spp. contained either tet(A/M/S) resistant genes as follows: tet(A) in A. veronii/sobria (n = 1), A. media (n = 2), A. aquariorum (n = 1), and A. veronii (n = 3); tet(M) in one isolate of A. sobria and tet(S) in 1 isolate of A. jandaei. In other bacteria, tet(A) gene was detected in Citrobacter freundi (n = 1), Pseudomonas putida (n = 1); tet(S) was also identified in Yersinia ruckeri (n = 1), Arthrobacter arilaitensis (n = 1) and P. putida (n = 1). In total, 31 isolates (31.00%) contained the tetracycline resistance genes in which 21 bacteria (21.00%) showed the tet(S), nine bacteria (9.00%) contained the tet(A) and 1 bacteria (1.00%) was positive for tet(M). All of the L. garvieae isolates contained tet(S) in this study. The most widely distributed resistance gene was gene tet(A) and the least known resistance genes was tet(M) among the studied bacteria of the genus Aeromonas in this study.

The description and host-parasite relationships of a new quadrigyrid species (Acanthocephala) from the Persian tooth-carp, Aphanius farsicus (Actinoptreygii: Cyprinodontidae) in Iran.

Acanthogyrus (Acanthosentis) barmeshoori n. sp. (Quadrigyridae) is described from the Persian tooth-carp, Aphanius farsicus Teimori, Esmaeili, and Reichenbacher, 2011 (Cyprinodontidae) in the Maharlu Lake basin, southern Iran. Aphanius farsicus is an endemic freshwater fish found in streams and springs that drain into Maharlu Lake, Shiraz, Iran. The new species is the smallest of all the 44 known species of the subgenus Acanthosentis Verma and Datta, 1929, measuring between 0.26 and 1.68 mm in length. It is further distinguished by having a short cylindrical proboscis with very long anterior hooks widely separated from very small hooks in 2 very close circles posteriorly (hook length ratio about 4:1). It is separated from 4 other species of Acanthosentis with similar proboscis armature but with less-extreme diversification of hook length. The new species is also distinguished in having anterior para-receptacle structures (PRS) and a similar posterior structure like those reported in only 1 other species of Acanthosentis from Japan. Proboscis receptacle is single walled with a large triangular cephalic ganglion. Testes are large, pre-equatorial, and Saefftigen's pouch is prominent. Fourteen to 25 circles of spines cover the anterior 50-70% of the trunk, but a few spines may be present at posterior end of trunk. This is the first species of Acanthosentis where SEM images, showing external morphological details, are provided. From a total of 357 fish specimens examined between July 2006 and June 2007, 173 specimens (48.5%) were infected with individuals of the new species. The prevalence of infection decreased with increasing fish size. The parasite was observed all year, with the highest abundance and intensity in May while the prevalence was highest in February. The prevalence of acanthocephalans decreased with increasing fish size. While most worms were recovered in fish within the length range of 18-29.9 mm, 1 of the longest parasites (1.68 mm long) was found in fish within the range of 30-33.9 mm long.