Chalcalburnus tarichi is an endemic cyprinid fish living in the Lake Van basin located in the Eastern Turkey. Fish (3+ ages) were exposed to 17α-ethynylestradiol (EE(2); 1, 10, 100 ng L(-1); nominal concentrations) and nonylphenol (NP; 10, 60, 200 μg L(-1) ; nominal concentrations) for 32 days under semistatic daily renewal conditions. The exposure period was followed by an evaluation of liver and gonadal apoptosis and gonad histopathology in males and females. Exposure to the highest concentrations of EE(2) (100 ng L(-1)) and NP (200 μg L(-1) ) caused significant increases in the extent of apoptosis in liver and gonads. Treatment with 100 ng L(-1) of EE(2) and 200 μg L(-1) NP increased the number of TUNEL positive hepatocytes significantly in both sexes compared to controls. The rates of apoptosis in testicular germ cells and ovarian follicular cells were significantly greater at the same concentrations. Exposure to EE(2) (100 ng L(-1)) and NP (60 and 200 μg L(-1)) caused thickening of interstitial connective tissue (fibrosis) in the seminiferous tubule wall and testis-ova formation in males. In females treated with 100 ng L(-1) EE(2) , increased percentage of atretic ooctytes and fibrotic areas in the ovarian somatic stromal tissue were found in the ovaries. Increase in atresia, without a statistical significance, and fibrotic stromal tissue were also noted in 60 and 200 μg L(-1) NP treatments. Results suggest that EE(2) - and NP-dependent hepatotoxicity and gonadotoxicity are causally related to the increase in apoptosis in C. tarichi.
The gonad histology, gonado-somatic index (GSI), 17b-estradiol (E 2 ) levels and acetylcholinesterase (AChE) activity in the carp species Chalcalburnus tarichi from Lake Van and the Karasu river, eastern Turkey, have been investigated. Fish between 5 and 7 years old were sampled from November 2003 to February 2004. The ratio of female fish caught in Lake Van with abnormal ovaries (AbOF) was 43.3%, but the fork length and body weight of these fish were not correlated with this abnormality. The weight of the ovaries and the GSI values of AbOF were very low (P < 0.05). Histological observations on the samples caught each month revealed that the oocytes had degenerated in the perinucleolus and early cortical alveolus stages and that the ovaries were full of somatic stromal tissue. In addition, the seminiferous tubules of male fish with abnormal testes did not contain male reproductive cells at any stage. The ovaries of the fish caught from the Karasu river were also full of oocytes in the perinucleolus and early cortical alveolus stages, but there were fewer atretic follicles. Furthermore, apoptosis was observed in the ovary cells of these fish, in particular in the follicular cells, and the plasma E 2 levels of the AbOF was very low (P < 0.05). AChE activity was inhibited only in liver (P < 0.05). We conclude that our sample of C. tarichi must have been exposed to various polluting chemicals or another unknown factors (such as global warming) and that these factors have irreversibly impaired oocyte development in a high percentage of fish.
In the present study, the testis histology, gonadosomatic index (GSI), germ cell proliferation and apoptosis, and the plasma 11-ketotestosterone (11-KT) and testosterone (T) levels of male Chalcalburnus tarichi were analyzed. According to the histological examinations of the specimens that were caught between February 2009 and January 2010, three testicular stages were determined. Those stages were as follows: (1) recrudescence or prespawning (July-April), (2) spawning (May-June), and (3) postspawning (July). It was observed that the GSI increased gradually, starting from the recrudescence stage, and it reached peak values at the spawning stage, while the lowest values were in the postspawning. Germ cell proliferation in the testis was detected using a proliferating cell nuclear antigen (PCNA), and germ cell apoptosis was detected by transferase dUTP nick end labeling staining. The germ cell PCNA and apoptosis index values were calculated. It was indicated that germ cell proliferation was observed in all of the testicular stages. The highest germ cell PCNA index (PI) levels were detected in July, August, and September, which then dropped in October and stabilized between February and April. The lowest PI values were detected in the spawning stage (May-June). Germ cell apoptosis was observed in all of the months, and the highest apoptotic index values were detected in August, September, October, May, and June. Plasma 11-KT and T levels were at their highest levels in May and June, and it was detected as stabile in the other months. There was a correlation between GSI, PI, and plasma androgen levels. In conclusion, the present data illustrate testicular development stages for C. tarichi and show changes in the level of GSI and sex steroid biosynthesis through spermatogenesis.
Recent reports have demonstrated gonadal abnormalities in the Lake Van fish (Alburnus tarichi) from Lake Van caused by increasing pollution. In the present study, the fish was collected from an area of Lake Van receiving mainly sewage treatment plant effluent at prespawning period (April) and from a river (Karasu) which is close to the polluted area of the lake and where the fish migrates at spawning period (May). Collected specimens were examined for testicular alterations, gonadosomatic index (GSI), condition factor (CF), and antioxidant defense system biomarkers based on comparison with a reference lake (Erçek) and a reference freshwater inlet (Memedik River). Histological examinations of the testes of fish from the polluted area and the connected river showed various alterations consisting of macrophage aggregates, vacuolation, pyknosis, germ cell degeneration, seminiferous tubule dilation, disorganization of tubules, reduced spermatozoa, and fibrosis. A lower GSI and CF were also observed. Moreover, alterations in the antioxidant system biomarkers were determined in the testis tissues of fish from the Lake Van and Karasu River, indicating oxidative stress. These results suggest that the abnormalities in the testes are causally related to the increased oxidative stress, and pollution in Lake Van may have adversely affected the reproductive health of the lake Van fish.
Bisphenol F (BPF) has been used frequently in the plastics industry and the production of daily consumer products as an alternative to bisphenol A (BPA). It was aimed herein to determine the cytotoxic effects of BPF on hepatocytes isolated from the liver of rainbow trout (Oncorhyncus mykiss) using lactate dehydrogenase (LDH) assay and antioxidant defence system indicators. The cultured hepatocytes were exposed to seven concentrations (0, 15.63, 31.25, 62.50, 125, 250, and 500 µM) of BPF for 24 h. According to the LDH assay, the percentage of cytotoxicity was increased dose dependently in the cells.The malondialdehyde content, which is indicative of lipid peroxidation, was increased signi cantly at BPF concentrations between 15.63 and 250 µM, whereas it remained unchanged with a concentration of 500 µM. The activities of superoxide dismutase were increased, while those of catalase were decreased with all of the BPF concentrations. Elevated levels of reduced glutathione content were determined with BPF concentrations between 15.63 and 250 µM, but decreased signi cantly with a concentration of 500 µM.Signi cant increases in the activities of the glutathione peroxidase were found in hepatocytes treated with BPF at concentrations of 31.25 to 500 µM. GST activity was only signi cantly increased with a BPF concentration of 250 µM. The results showed that the toxic mechanism of BPF was mainly based on cell membrane damage and oxidative stress, which have an in uence on antioxidant defences. Therefore, BPF was reconsidered as a safe alternative instead of BPA in the manufacturing of industrial or daily products.
In this study, isolated hepatocytes of pearl mullet (Alburnus tarichi) were exposed to bisphenol A (BPA) at concentrations of 25, 50, 100, and 200 µM for 24 h. Moreover, an in vitro antioxidant concentration of vitamin C (50 µM) was administrated to the culture medium along with the BPA exposures. Next, the antioxidant defense system parameters were analyzed. According to the results, the highest concentration of BPA (200 µM) proved to be severely toxic for the cells. The increased activities of superoxide dismutase (SOD) and glutathione-S-transferase (GST), the fluctuated activities of glutathione peroxidase (GPx), and the decreased content of reduced glutathione (GSH) were compared to the control group after the BPA exposures. Vitamin C co-administration was found to cause further increases in the SOD, GPx, and GST activities in some of the experimental groups and vitamin C could not restore the GSH content. Malondialdehyde (MDA) levels were observed to be unaffected in all exposure groups. These results show that BPA causes alterations in the antioxidant defenses of the isolated fish hepatocytes. In addition, vitamin C co-administration along with BPA was found to be non-protective against BPA-induced oxidative stress, consequently, aggravated a negative BPA impact.
The Lake Van fish (Alburnus tarichi) is a species that is endemic to Turkey’s Lake Van basin. In this study, the regional distribution, volume density, and relative frequency of some pancreatic endocrine cells in Lake Van fish were investigated via immunohistochemistry using specific mammalian antibodies. The pancreatic tissue was observed to be surrounded by adipose tissue, which was adjacent to the gall bladder or extrahepatic bile duct, or dispersed in the adipose tissue ranked among coils of post-esophageal swelling and intestine. The pancreatic endocrine cells were examined, including the islets, exocrine pancreas, and pancreatic ducts. According to the modified aldehyde fuchsin staining and immunohistochemistry, insulin-secreting beta cells were observed to localize throughout the islets. Glucagon immune-reactive (IR) cells were observed to be situated moderately on the islet periphery, and were rarely determined in the islet central region. A small number of somatostatin-IR cells were observed in the islet centers and peripheries. Similar distributions of those 3 endocrine cells were also determined in the secondary islets. Additionally, the endocrine cell percentages did not differ between the primary and secondary islets; insulin-, glucagon- and somatostatin-IR cells comprised approximately 54%, 29%, and 11% of the endocrine cells in the principal islets, whereas they comprised 52%, 27%, and 14% in the secondary islets, respectively. Insulin-, glucagon- and somatostatin-IR cells were also determined among the epithelium and subepithelial connective tissue in the pancreatic ducts or exocrine areas of the pancreas. With this study, the existence, regional distribution, and relative frequency of the insulin-, glucagon- and somatostatin-IR cells were first investigated in the pancreatic tissue of Lake Van fish and the results were discussed.
Alburnus tarichi is an endemic fish species inhabiting the Lake Van basin of Turkey. This anadromous cyprinid fish migrates for spawning to the freshwater inlets pouring into Lake Van, which has highly alkaline and brackish water. During the migration, the fish moves to a new habitat from a different habitat and encounters many challenges. The objective of the present study is to investigate whether antioxidant responses occur in the tissues of A. tarichi during its anadromous migration. To this end, fish were sampled at three different points and in two different periods from the migration route, including in Lake Van at prespawning, at the entrance of a freshwater stream (Karasu) at spawning and in the upstream of the freshwater stream at spawning. Malondialdehyde (MDA) content and antioxidant defenses including activities of the enzymes superoxide dismutase, glutathione peroxidase, glutathione-S-transferase and catalase and the level of glutathione were assayed in liver, gill, white muscle, trunk kidney and anterior intestine tissues. Our results showed increased MDA levels in liver, white muscle and anterior intestine and tissue-specific antioxidant responses in the freshwater environment. This study reports that alterations occurred in the antioxidant defense system indicators in the tissues of anadromous cyprinid fish during migration and that the antioxidant defenses might reflect an important role in spawning migration that ultimately leads to accomplishment of reproductive activity.
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