The aim of this study was to identify the effects of dithiopyr (DTP), a herbicide, on behavior in zebrafish. The toxicity of DTP has rarely been investigated in fish. In the present study, zebrafish were exposed to different concentrations of DTP in the range of 10-20 μM for 48 h in a test container, in order to measure the value of median lethal concentrations (LC50). Behavioral experiments were performed, including the novel tank test (NTT) and the open field test (OFT), to assess stress responses or locomotion. After exposure to the DTP solution at a sublethal concentration of 2.5–10 μM for 6 min, the behavior of the zebrafish was observed for 6 min. In the acute toxicity test, the LC50 value of DTP showed as 14.49 μM in the zebrafish. The NTT showed that the duration of immobility and the velocity were significantly increased by exposure at a concentration of 5 μM of DTP, compared with a control group (p<0.05). However, compared with the control group, DTP significantly decreased the distance moved and the frequency at the top of the tank, and significantly increased the turn angle and duration at the bottom, in a concentration-dependent manner (p<0.05). In addition, in the OFT, exposure to DTP significantly decreased the distance moved and velocity compared with the control group (p<0.05). Exposure to DTP also significantly increased the duration of immobility, the turn angle, and the meandering movement, in a concentration-dependent manner (p<0.05). Further, exposure to DTP at a low concentration elevated whole-body cortisol levels in the zebrafish. The results of this study thus suggest that DTP induces a toxic response and negative effects on behavior and the endocrine system in zebrafish.
Many of studies on the transovarial transmission of occult virus and their activation due to various stresses such as cold or heat treatment, chemical feeding, and nutritional deficiency, etc., in the silkworm, Bombyx mori L. have been made, but any attempts have been not made to control virus diseases by detection of the occult virus-carried moths in the production of silkworm egg of hybrids, because of difficulty to detect occult virus in any stage. Therefore, it may be worth while to disclose whether a sublethal infection of the moths from which active virus are detectable, has the same level of induction rate as that of occult virus activation, thus to apply its results for the reduction of the occurence of virus diseases in silkworm rearing. For these purposes, the following experiment was conducted as one of preliminary steps. In this study, investigations on the generation-to-generation transmission of occult virus and a sublethal infection, and the role of chromosomal gene of the host, Jam 103 and Jam 104 in the Previous generation, and Jam 103 x Jam 103 and Jam 104 f Jam 104 in the next generation were made for the induction of virus diseases due to the transmitted virus. The frequency of cytoplasmic polyhedrosis due to the induction in the F$_1$ generation was markedly higher in the cross-batches, male$\times$female and male$\times$female in which inoculated individuals were used as fem ale parents than in the cross-batches, male$\times$female and male$\times$female in which virus has been not inoculated or inoculated only to male in the previous generation. The tendency of increasing rate was observed in any treatments; such as the inoculations of cytoplasmic polyhedrosis virus (10$\^$5/, 10$\^$6/ 10$\^$7, and 10$\^$8//ml ill different concentration of inocula) , cold-treatment (5$^{\circ}C$, 12hrs or 24hrs), and formalin-feeding treatment (2% or 3%). The shape of polyhedra (tetragonal in outline) examined in the F, larvae was identified as that of the inoculated polyhedra with partial application of immunofluorescent techniques. These results suggests that the cytoplasmic polyhedrosis virus in B. meri L. are transmitted to the next generation through the egg, apparently in the occult state. And the experimental results of various cross-batches revealed the egg cytoplasm plays an important part i the transmission of the occult virus of the cytoplasmic polyhedrosis virus,
The selective tox\ulcornercity of fenpyroximate to the predatory mite Amblyseius womersleyi and the twospotted spider mite Tetranychus urticae was evaluated. Adult females and eggs of both species were placed on bean leaf dis~sd ipped in several concentrations of fenpyroximate. Fenpyroximate was much less toxic to A. womersleyi than to T. urticae. Although the survival of adult females of A. womersleyi tended to decrease with increasing fenpyroximate concentration, 58-74% of predators remained alive at concentrations of 6.25-50 ppm. However, reproduction of predators was not significantly reduced at any of the concentrations tested. At 6.25-50 ppm, 32-40% of twospotted spider mite adult females survived but all survivors were immobilized. Moreover, reproduction of twospotted spider mites was reduced with increasing fenpyroximate concentration. Fenpyroximate did not affect the hatch of A. womersleyi eggs or the development of immature predators. Although survival of immature predators decreased with increasing fenpyroximate concentration, 16-48% of immature predators reached adulthood at 6.25-50 ppm. However, all immature spider mites failed to develop to adulthood at 6.25-50 ppm. Adult female predators survived on a diet of twospotted spider mites intoxicated with fenpyroximate, and their fecundity and sex-ratio of the progeny were not substantially affected. Fenpyroximate at selective sublethal concentrations (6.25-12.5 ppm), therefore, could be of value in adjusting predatorlprey ratio in integrated management of twospotted spider mites.
Mercury (Hg) is a major concern in marine environment because of their bioaccumulation and biomagnification properties, and adverse effects to aquatic organisms at even a trace amount. However, little information on the effects of Hg, compared to other heavy metals, is available in marine small crustaceans. Here, we investigated the transcriptional modulation of metabolism-related genes in the brackish water flea, Diaphanosoma celebensis after exposure to sublethal concentration (0.2, 0.4, 0.8 ㎍/l) of HgCl2 for 48 h. Relative mRNA expression levels of five detoxification enzyme-coding genes (cytochrome P450; cyp360A1, cyp361A1, cyp4AP3, cyp4C122, and cyp370C5) and six digestive enzyme-coding genes [alpha amylase (AMY), alpha amylase related protein (AMY-like), trypsin (TRYP), chymotrypsin-like protein (CHY), lipase (LIP), pancreatic lipase-related protein (PLRP)] were analyzed using quantitative real time reverse transcription polymerase chain reaction (qRT-PCR). As results, Hg increased the mRNA level of cyp370C5 (clan2) and cyp4AP3 (clan4) in a concentration dependent manner. A significant increase in TRYP mRNA was also concentration-dependently observed after exposure to Hg. These findings suggest that cyp370C5 and cyp4AP3 play a key role in Hg detoxification in D. celebensis, and Hg can affect energy metabolism by modulating the transcription of digestive enzyme. This study will provide better understanding the molecular effects of Hg in marine small crustacean.
Ahn, Youngbeom;Kim, Jeong Myeong;Lee, Yong-Jin;LiPuma, John J.;Hussong, David;Marasa, Bernard S.;Cerniglia, Carl E.
Journal of Microbiology and Biotechnology
/
v.27
no.12
/
pp.2211-2220
/
2017
Chlorhexidine gluconate (CHX) and benzalkonium chloride (BZK) formulations are frequently used as antiseptics in healthcare and consumer products. Burkholderia cepacia complex (BCC) contamination of pharmaceutical products could be due to the use of contaminated water in the manufacturing process, over-diluted antiseptic solutions in the product, and the use of outdated products, which in turn reduces the antimicrobial activity of CHX and BZK. To establish a "afe use" period following opening containers of CHX and BZK, we measured the antimicrobial effects of CHX ($2-10{\mu}g/ml$) and BZK ($10-50{\mu}g/ml$) at sublethal concentrations on six strains of Burkholderia cenocepacia using chemical and microbiological assays. CHX (2, 4, and $10{\mu}g/ml$) and BZK (10, 20, and $50{\mu}g/ml$) stored for 42 days at $23^{\circ}C$ showed almost the same concentration and toxicity compared with freshly prepared CHX and BZK on B. cenocepacia strains. When $5{\mu}g/ml$ CHX and $20{\mu}g/ml$ BZK were spiked to six B. cenocepacia strains with different inoculum sizes ($10^0-10^5CFU/ml$), their toxic effects were not changed for 28 days. B. cenocepacia strains in diluted CHX and BZK were detectable at concentrations up to $10^2CFU/ml$ after incubation for 28 days at $23^{\circ}C$. Although abiotic and biotic changes in the toxicity of both antiseptics were not observed, our results indicate that B. cenocepacia strains could remain viable in CHX and BZK for 28 days, which in turn, indicates the importance of control measures to monitor BCC contamination in pharmaceutical products.
The goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals. Japanese medaka was treated by copper of appropriate sublethal concentrations after starvation for 48 hr. The untreated individuals showed common behavioral characteristics (i.e. , smooth and linear movements). Locomotive activity of the fish was monitored using an image processing and automatic data acquisition system. When treated with copper (100 ppb), the fish showed shaking patterns more frequently. As the concentration of copper increased to 1,000 ppb, activity decreated, and the fish showed an erratic movement. Fish were exposed to copper at various concentrations (0,100 and 1,000 ppb) for 24 hrs, and acetylcholine esterase (AChE) activity was observed. When fish were exposed to 1,000 ppb of copper, the body AChE activities appeared to decrease but the head AChE activities showed little change. Expressions of tyrosine hydroxylase (TH) protein in the different organs from both head (brain) and body (kidney) portions affected by the copper treatment were analyzed using immunohistochemical technique compared with control. Five organs of the fish (olfactory bulb, hyothalamus, optic lobe, pons and myelencephalon regions) showed a relatively strong TH protein expression in the control experiment. A differential expression of TH, however, was observed in the treatment (100 ppb and 1,000 ppb). The treatment (1,000 ppb) significantly suppressed TH protein production in the brain regions. In kidney, however, the same treatment caused little suppression compared with the control. Copper appeared to be less effective in suppression of TH than diazinon, a known TH suppressor. It was concluded that TH could be used at a potential biomarker to monitor the acute copper toxicity in Japanese medaka.
The investigation of the Acetylcholinesterase (AChE) activity in tissues (brain, eye, muscle and serum) of crucian carp (Carassius auratus) exposed to the waterborne parathion was carried out for application as biomarker of organophosphate pesticides. The AChE activities were significantly inhibited in the experimental organs of C. auratus treated ${\geq}63{\mu}g/L$ of concentrations of parathion. The AChE activity of C. auratus was significantly reduced in response to brain (79.1~92.4%), eye (76.0~91.5%), muscle (89.7~97.6%) and serum (68.9~78.0%) after 30 days exposure. No significant mortality occurred during the experiment duration but behavioral changes occurred in the carp after exposure to the parathion were erratic swimming and convulsions. The anomaly in the carp exposed to parathion were observed in the form of scoliosis. The use of AChE activity and other adverse responses of the carp might be use as a reliable monitoring tool to detect parathion in aquatic ecosystem which might produce significant population changes.
Sublethal dose of bacterial lipopolysaccharide (LPS) would induce protection against cardiac ischemic/reperfusion (I/R) injury. This study examines the following areas: 1) the temporal induction of the cardio-protection produced by LPS; and 2) the relations between a degree of protection and the myocardial prostacyclin ($PGI_2$) production. Rats were administered LPS (2 mg/kg, i.v.), and hearts were removed 1, 4, 8, 14, 24, 48, 72,and 96 h later. Using Langendorff apparatus, haemodynamic differences during 25 min of global ischemia/30 min reperfusion were investigated. The concentration of $PGI_2$ in aliquots of the coronary effluent was determined by radioimmunoassay as its stable hydrolysis product $6-keto-PGF1_{\alpha}$ and lactate dehydrogenase release were measured as an indicative of cellular injury. LPS-induced cardiac protection against I/R injury appeared 4 h after LPS treatment and remained until 96 h after treatment. $PGI_2$ release increased 2-3 fold at the beginning of reperfusion compared to basal level except in hearts treated with LPS for 48 and 72 h. In hearts removed 48 and 72 h after LPS treatment, basal $PGI_2$ was increased. To determine the enzymatic step in relation to LPS-induced basal $PGI_2$ production, we examined prostaglandin H synthase (PGHS) protein expression, a rate limiting enzyme of prostaglandin production, by using Western blot analysis. LPS increased PGHS protein expression in hearts at 24, 48, 72, 96 h after LPS treatment. Induction of PGHS expression appeared in both isotypes of PGHS, a constitutive PGHS-1 and an inducible PGHS-2. To identify the correlationship between $PGI_2$ production and the cardioprotective effect against I/R injury, indomethacin was administered in vivo or in vitro. Indomethacin did not inhibit LPS-induced cardioprotection, which was not affected by the duration of LPS treatment. Taken together, our results suggest that $PGI_2$ might not be the major endogenous mediator of LPS-induced cardioprotection.
This study was carried out to estimate toxic effects of phenol on survival and metabolism of the abalone juvenile, Haliotis discus hannai. The experiment was conducted by renewal bioassay procedure with different salinities at $20^{\circ}C$. The $LC_{50}$ of the juvenile exposed to phenol in the range of 0.5 and $100mg/\ell\;was\;34.3\~6.5mg/\ell\;at\;2.4\%_{\circ}\;and\;52.2\~9.3m/\ell\;at\;32\%_{\circ}$ salinity with exposure time from 24 hours to 96 hours. $LT_{50}$ was remarkablely reduced with increase of phenol conentration and decrease of salinity. Lethal toxicity or phenol was higher at low salinity than at high salinity. Therefore, salinity is likely to be one of factor to increase phenol toxicity. The oxygen consumption of the juvenile was reduced with increase of phenol concentration and with decrease of salinity. In spite of phenol toxicity, the oxygen consumption of the juvenile exposed to phenol of low concentration was high and similar as compared with that of control group. Survival rates of the abalone kept in phenol-free sea water after exposure to phenol concentration of 5, 10 and $20mg/\ell$ for 96 hours were reduced with decrease of salinity. Durations required to recover the normal metabolic rate of the juvenile, which was exposed to phenol concentration of 5, 10 and $20mg/\ell$ for 96 hours, were made longer with increasing phenol concentration. In the case of the juvenile exposed to sublethal concentration of phenol for 15 days, it were elongated as compared with that of the abalone exposed to phenol concentration caused acute toxicity. The result of this experiment indicated that relatively low concentration of phenol can impact on the abalone juvenile in marine ecosystem.
The aim of this work was to investigate the cellular responses and proteomic analysis of Bacillus cereus MH-2 exposed to EGCG. Strain MH-2 was isolated from commercial Ssamjang and has the hemolytic activity. Survival of the MH-2 strain with time in the presence of different concentrations of EGCG under sublethal conditions was monitored. The amount of alginate from MH-2 strain decreased depending on the increasing concentrations of EGCG and increased depending on the exposure time at any particular EGCG concentration. Analysis of SDS-PAGE and Western blot using anti-DnaK and anti-GroEL revealed that two stress shock proteins, 70 kDa DnaK and 60 kDa GroEL were found to decrease in proportion to the EGCG concentration in exponentially growing cultures. Scanning electron microscopic analysis demonstrated the presence of protrusions and fused rod forms on the cells treated with EGCG. 2-DE of soluble protein fractions from MH-2 cultures showed 20 protein spots changed by EGCG exposure. These proteins involved in enterotoxins (hemolysin BL lytic component L1 and hemolysin BL-binding protein), chaperons (DnaK and GroEL), cell defense (peptidase M4 family proteins), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. These results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on B. cereus MH-2.
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