Alburnus Tarichi Research Articles

Determination of cytotoxic, genotoxic, and oxidative damage from deltamethrin on primary hepatocyte culture of Lake Van fish, Alburnus tarichi

ABSTRACT Deltamethrin is a widely used pyrethroits worldwide. Although the chemical is used to combat insects, it has effects on other non-target organisms. Deltamethrin is extensively used in agriculture, animal husbandry, and domestic areas in the Lake Van basin. The aim of this study was to determine the cytotoxic, genotoxic, and oxidative damage from deltamethrin on the primary hepatocyte culture of Lake Van fish (Alburnus tarichi). In this study, the toxic effects of different concentrations (0.01, 0.1, 1, and 10 µM) of deltamethrin in the primary hepatocyte culture of Lake Van fish were investigated via liver enzymes aspartase aminotransferase (AST) and alanine aminotransferase (ALT), and the total antioxidant status (TAS), total oxidant status (TOS), lipid peroxidation (MDA), and DNA damage (8-OHdG). Deltamethrin caused an increase in the AST and ALT levels dependent on the dosage and time. The TAS and TOS levels increased at the end of 24 h and there was no difference at the end of 48 h. Deltamethrin did not affect the MDA level, but increased the 8-OHdG (P < 0.05). In conclusion, it can be said that high doses of deltamethrin (1 and 10 µM) have a toxic effect on the primary hepatocyte culture of Lake Van fish.

Roe protein hydrolysate of Alburnus tarichi induces apoptosis in breast cancer MCF-7 and MDA-MB-231 cells through a caspase-dependent pathway.

The protein hydrolysates of fishes have been reported to be a potential source of many health benefits components for pharmaceutical or nutritional applications. The aim of this study is to examine the possible antiproliferative function of roe protein hydrolysates of Alburnus tarichi using enzymatic hydrolysis against breast cancer cells and explore its detailed mechanisms. In addition, we evaluated the effects of protein hydrolysate on the proliferation and apoptosis of two human breast cancer cell lines (MCF-7 and MDA-MB-231). The cultured cells were treated with protein hydrolysate at concentrations of 0-5 μg/ml for 24 h and 48 h. Inhibition of cell proliferation, percentage of apoptotic cells, cell cycle distribution, morphological changes, DNA fragmentation, intracellular reactive oxygen species (ROS) production, and apoptotic protein levels were also examined. Decreases in proliferation of MCF-7 and MDA-MB-231 cells were observed after treatment with the protein hydrolysate in a dose-dependent manner. Distinct morphological changes, a typical pattern of fragmented DNA, and increased intracellular ROS production and apoptotic protein levels were observed in both cell lines after hydrolysate treatment (p < 0.05). The results suggested that the protein hydrolysate inhibits the proliferation of human breast cancer cell lines by introducing apoptosis through a caspase-dependent pathway in a dose-dependent manner.

Histopathology and immunohistochemistry of gills of Van fish (Alburnus tarichi Güldenstädt, 1814) infected with myxosporean parasites

ABSTRACT The Van fish are a cyprinid species endemic to Turkey’s largest soda lake, Lake Van, and have great economic value because they are a food source. Once a year, the fish take part in reproductive migration to the fresh waters flowing into the lake. The fish migrate from an extreme environment with high salinity (2.2%) and high pH (9.8). These fish are unable to reproduce in this alkaline environment and must migrate to fresh water during their breeding season. The aim of the present study is to report the presence of the myxosporean parasites on the gills and the pathological changes. Changes in gill histopathology, mucocytes, mitochondria-rich cells, expression of Heat Shock Protein 70 (Hsp70), and ATPase (NKA) were observed in the gill tissue. As a result of the histopathological changes in gills, infected fish had abundant plasmodia with different sizes. Plasmodia were found on gill filaments inside white ovoid-shaped structures. It was observed that plasmodia were contained on the primary filament which changed the histological structure of the gill tissue to a large extent. It was determined that the density and size of mucocytes in the infected areas of the gill tissue increased, whereas the number of mitochondria-rich cells decreased. Hsp70, an indicator of stress, was not different between normal and infected fish.

Polyclonal antibody-based immunoassay of vitellogenin in Van fish (Alburnus tarichi)

ABSTRACT Van Lake is the third largest closed lake in the world and the biggest lake in Turkey. An ELISA method has developed with the aim of determining the pollution caused by estrogens and estrogen-like chemicals that have come to the lake Van in recent years. First, the vitellogenin in estrogen-treated male fish plasma was purified by ion exchange chromatography, injected into rats, and the obtained polyclonal antibodies were tested for specificity by Western blot and immunohistochemical methods. Immunohistochemical labeling of the vitellogenin-synthesized liver resulted in the intense marking of the liver of the animals injected with estrogen, while no markings were observed in the control group. The limit of detection of the developed enzyme-linked immunosorbent assay was 4.6 µg L−1, and the working range was 7.8 to 2000 µg L−1. Intra- and inter-assay variations were 13.0 % and 13.3%. The highest level of vitellogenin in male fishes measured was 23.56 µg mL−1.

Anesthetic Activity of Clove Oil (Eugenol) on the Lake Van Fish (Chalcalburnus tarichi Pallas, 1811)

The use of clove oil (eugenol) as an anesthetic for Lake Van fish(Chalcalburnus tarichi), an endemic cyprinid fish species ofLake Van basin, was examined. Induction times were determined to be ≥35, &amp;lt;4, &amp;lt;3, &amp;lt;2, &amp;lt; 2 and ≥1 min at 20, 40,50, 60, 80 and 100 mg/L clove oil concentrations, respectively. Recovery times upon 10 min of exposure to eugenol were determined respectively to be &amp;gt;3, &amp;gt;4,&amp;gt;3, &amp;gt;4,&amp;gt;6 and &amp;gt;8 min clove oil can be considered as a suitable anesthetic for Chalcalburnus tarichi and an optimum anesthetic concentration of 50 mg/L is recommended.

Microbiological and chemical quality of different types of salted pearl mullet (Chalcalburnus tarichi Pallas, 1811)

AbstractThis research aimed to determine microbiological and chemical quality of different types of salted pearl mullet (Chalcalburnus tarichi Pallas, 1811). Samples of salted pearl mullet (C. tarichi Pallas, 1811), produced by six different methods, were obtained from three different local producers at the end of the hunting season of 2016 and 2017 (May–June) with wooden boxes, plastic cans, or in bottom perforated plastic cans. Samples stored in a cold storage (2 ± 2°C) were analyzed in August and September of 2017. Six fish were used in each sampling day and thus the analyses were completed in 18 days. So, 108 fish samples were analyzed (3 producer × 6 method × 3 sample for each method × 2 hunting season). Analysis results were calculated as mean ± SD. One‐way analysis of variance was used to determine whether there was a significant difference between the sample groups analyzed, and p &lt; .05 was considered significant. All sample groups were found to be appropriate according to total mesophilic aerobic bacteria count, but one sample group (OS11‐old sample, gutted and cleaned fish producted in wooden box according to method 1) included Salmonella spp. and Vibrio parahaemolyticus, it was found to be inappropriate (p &lt; .05). The total volatile basic nitrogen acceptability upper limit (25–30 mg/100 g) were found to be exceeded in three sample groups (OS12‐old sample, ungutted, and uncleaned fish producted in wooden box according to method 1, OS31‐old sample, gutted, and cleaned fish producted in bottom perforated plastic can according to method 3, OS32‐old sample, ungutted, and uncleaned fish producted in bottom perforated plastic can according to method 3) (p &lt; .05) and the tiobarbituric acid index value was found to vary between 13.32 ± 0.08 and 31.43 ± 0.04 μg malondialdehyde/g (p &lt; .05). According to histamine results, all sample groups were determined at acceptable values (p &gt; .05). However, it was determined that the salt amount of the sample groups was very high in raw, and that more than half of the salt amount remained in the samples, even after baked waiting for 3 hr in warm water (p &lt; .05).

Effects of cypermethrin on antioxidant enzymes and lipid peroxidation of Lake Van fish (Alburnus tarichi)

Lake Van fish is the only vertebrate species that lives in extreme condition (salinity of 2.2%, alkalinity of 151.2 meq/kg, and pH of 9.8) of largest soda lake of the world. This fish is an important source of protein for the people living in Van and surrounding provinces. . Cypermethrin (CYP) is used extensively against insects in agricultural, livestock, and domestic applications in Lake Van basin. CYP has toxic effects highly on non-target organisms as aquatic animals. The aim of this study was to evaluate the effect of commercial formulas of CYP (0.05 µg/L) on Van Lake fish (Alburnus tarichi) for 24, 48, 72, 96 h under semi-static conditions using oxidative damage biomarkers. In this study, antioxidant defense system enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) activities and lipid peroxidation (LPO) product malondialdehyde (MDA) levels were determined in the liver, kidney, gills, and brain tissues. According to the results of this study, the decrease in SOD (at 48, 72, and 96 h) and CAT and GPx (at 24, 48, 72, and 96 h) enzyme activities were significant with exposure to CYP (p < 0.05). In addition, the increase in MDA levels, which is the indicator of LPO, was found to be significant (p < 0.05). However, in the 24 h period exposed to CYP, a significant increase in SOD activity was determined in kidney and brain compared to the control group (p < 0.05). In conclusion, sublethal concentrate application of CYP is thought to cause oxidative stress and disrupt the oxidant, antioxidant balance of Lake Van fish.

Variations in the lipid peroxidation and antioxidant biomarkers in some tissues of anadromous cyprinid fish during migration

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.

Van Balığında (Alburnus tarichi Guldenstadt, 1814) Üreme Göçü Sırasında Oksidatif Stresin Araştırılması

Van Fish is an anadromous species of endemic to Turkey's largest lake. The fish migrate to the freshwater pouring from the alkaline Lake Van to the lake for the reproduction every year. Fish are exposed to different stress factors such as fasting, salt and pH adaptation during migration. In this study, total oxidant and antioxidant, lipid peroxidation and DNA damages in tissues such as gill, liver, plasma, brain and muscle were compared before and after reproductive migration. Total oxidant status (TOS) of the fish caught in the lake before reproduction, liver and kidney tissues, total antioxidant status (TAS), fish liver caught from freshwater, fish from the lake were observed to be high in the brain and kidney tissues (P &amp;lt;0.05). Malondialdehyde (MDA) levels were found to be higher in the liver and kidney of the fish sampled from the freshwater and lake, respectively. (P &amp;lt;0.05). DNA damage was found only in the liver of fish sampled from freshwater. As a result, the reactions of the organs against stress effects encountered during reproduction migration were determined.

Immunohistochemical distribution of insulin-, glucagon- and somatostatin-containing cells in the pancreas of Lake Van fish (Alburnus tarichi Güldenstädt, 1814) (Cyprinidae).

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 three 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.

Apoptosis and cell proliferation in the epithelia of the esophagus and intestine of Alburnus tarichi Güldenstädt, 1814 (Cyprinidae) during migration from highly alkaline and brackish water to fresh water

In this study, cellular turnover in the epithelia of the esophagus and intestine of the anadromous Lake Van fish (Alburnus tarichi) that migrated from the highly alkaline and brackish water of Lake Van to fresh water were investigated by apoptosis and cell proliferation. Toward this aim, fish were collected from or near the entrance to Lake Van, and upstream in a freshwater stream (Karasu). The tissues were subjected to terminal deoxynucleotidyl transferase nick end labeling and proliferating cell nuclear antigen stainings for the detection of apoptosis or cell proliferation, and were then quantified. In the esophagus epithelium, the apoptotic index (AI) displayed no differences between fish from the lake and those from the freshwater stream. The AI increased significantly at the anterior or mid-intestine epithelia and throughout the intestine epithelium, whereas it remained unchanged in the posterior intestine epithelium of fish from both upstream and at the entrance of the freshwater stream. In the esophagus epithelium, the proliferation index (PI) values decreased gradually in fish from the freshwater stream. The PI increased significantly in the anterior intestine epithelium or throughout the intestinal epithelium of fish collected from the entrance of the stream; however, it did not change along the intestinal epithelium of fish collected upstream in the freshwater stream. It was determined that cellular changes occurred in the esophageal and intestinal epithelium of Lake Van fish during freshwater acclimation.

Oxidative and genotoxic effects of bisphenol A on primary gill cell culture of Lake Van Fish (Alburnus tarichi Güldenstädt, 1814)

ABSTRACTLake Van is the largest lake in Turkey. The lake limits lifespan due to its high pH and brackish water. For this reason, only a single species of fish (Van Fish) is living in the lake that has been adapted to these conditions. In the present study, we investigated the total oxidant status (TOS), total antioxidant status (TAS), malondialdehyde (MDA) level and DNA damage effect of bisphenol A (BPA) (10−7, 10−6 and 10−5 M) on primary gill cell culture of Van Fish for 24 and 48 h of incubation periods. TAS levels were not changed when compared to those of the control group, but TOS levels were decreased in both 24 and 48 h. The MDA level increased only at the highest concentration (10−5) at the end of 12 and 24 h (p < .05). DNA damage increased only at the 10−5 M concentration after 48 h. At the end of the experiment, BPA exposure caused lipid peroxidation and genotoxic effect. These results indicate that high levels of BPA exposure induced oxidative stress and DNA damage by time- and concentration-dependent fashion in the gill cell culture of Van Fish. Gill cell culture is a useful model for the rapid identification of the harmful effects of chemicals in the aquatic environment.

Antioxidant responses and DNA damage in primary hepatocytes of Van fish (Alburnus tarichi, Güldenstadt 1814) exposed to nonylphenol or octylphenol

Alkylphenols, a nonionic surface-active agent group, such as nonylphenol (NP) and octylphenol (OP) are important endocrine-disrupting chemicals (EDC). In this study, the dose- and time-dependent effects of NP and OP were investigated in the primary hepatocyte culture of Van Fish. In this study, samples were taken at different times and biochemical parameters were studied separately. The effects of the chemicals used on SOD, CAT, GSH-Px, MDA, and 8-OHdG were investigated in hepatocyte culture. The antioxidants SOD and CAT were observed to increase in all groups in the primary hepatocyte cultures at the 24th hour after NP and OP administration, whereas the GSH-Px level was observed to increase with OP at the 24th hour and with NP at the 48th hour. The MDA level was observed to reach its highest value for both chemicals in the 24th hour, and the 8-OHdG level was observed to increase toward the end of the follow-up time, compared to the control group (p < 0.05). In conclusion, different doses of NP and OP were found to induce an increase in the levels of antioxidants and the MDA level in Van Fish primary hepatocyte culture. DNA damage, on the other hand, may be considered to appear after longer-term exposure to NP and OP.

Inhibitory properties of some heavy metals on carbonic anhydrase I and II isozymes activities purified from Van Lake fish (Chalcalburnus Tarichi) gill.

In this study, CA I and II isoenzymes were purified from Van Lake fish gills by using Sepharose-4B-L-tyrosine-sulfanilamide affinity chromatography and to determine the effects of some metals on the enzyme activities. For purified CA I isoenzyme, yield, specific activity, and purification fold were obtained as 42.07%, 4948.12 EU/mg protein, and 116.61 and for CA II isoenzyme, 7%, 1798.56 EU/mg protein, and 42.38 respectively. Activity of CA was determined by measuring "CO2-hydratase activity". Purity control was checked by SDS-PAGE. In vitro inhibitory effect of Cu2+, Ag+, Cd2+, Ni2+ metal ions, and arsenic (V) oxide were also examined for both isozymes activities. Whereas Cu2+, Ag+, Cd2+, and Ni2+ ions showed inhibitory effects on both isozymes, arsenic (V) oxide showed activation effect. IC50 values were calculated by drawing activity %-[I] graphs for metal ions exhibiting inhibitory effects. IC50 values were determined as 3.39, 6.38, 13.52, and 206μM for CA I isozyme and 6.16, 20.29, 46, and 223μM for CA II isozyme respectively.

Hemato-biochemical responses of Van fish (Alburnus tarichi Guldenstadt, 1814) during sublethal exposure to cypermethrin

ABSTRACTCypermethrin is a highly active type II pyrethroid that is widely used to control harmful insects. The present study was conducted to investigate the possible effects of the sublethal concentration of cypermethrin (0.05 μL/L) on the biometric, hematological, and biochemical parameters of Van fish (Alburnus tarichi Guldenstadt 1814) for 24, 48, 72, and 96 h. Behavioral and physiological changes were observed in the fish treated with cypermethrin. Hematological parameters revealed a significant decrease in the erythrocyte (RBC), hemoglobin (Hb), and hematocrit (Hct) values (p ≤ .05). Serum enzymes, aspartate aminotransferase (AST, EC 2.6.1.1), alanine aminotransferase (ALT, EC 2.6.1.2), and lactate dehydrogenase (LDH, EC 1.1.1.27) activities and serum cortisol level were significantly increased (p ≤ .05). The results showed that changes in the biometric, biochemical, and hematological values of the Van fish exposed to cypermethrin can be used as biomarkers for monitoring toxicity.

Some macro and trace elements in various tissues of Van fish variations according to gender and weight

ABSTRACT Van fish is a fish that can live in soda water at pH 9-10. Van fishing area is a very important source of protein for the people. The variations in tissue metal levels according to gender and weight in Van fish (Alburnus tarichi, Güldenstadt 1814) were studied. This study was conducted for the first time in Van fish. Which it is very important for living organisms Calcium(Ca), magnesium(Mg), sodium(Na), potassium(K), cobalt(Co), chromium(Cr) and lithium(Li) elements levels in the muscle, liver, gill, gonad, and brain tissues of Van fish were investigated by using inductively-coupled plasma-optic emission spectroscopy (ICP-OES). The results were evaluated in two groups as macro-elements(Ca, Mg, Na, K) and trace-elements(Co, Cr, Li). Cobalt concentration in brain tissue was found to be higher than those in other tissues. Also, male fish had higher metal concentrations than female fish and their metal levels in brain and gonad tissues varied with their weight (P&lt;0.05). This study shows that consumption of Van fish can be consumed to supply some necessary minerals required for a healthy nutrition.

Effect of fish feed prepared with trout organs and Van Fish on the growth performance of Rainbow Trout

In this study, it was aimed to evaluate more efficiently Rainbow trout ( Oncorhynchus mykiss ) internal organs not consumed by humans and Van fish ( Alburnus tarichi , Güldenstädt 1814), which have low economic value. Easy digestible feed for Rainbow trout were prepared that is low cost and similar to natural food. The wastes were processed in an easy way with the semi-wet feed construction method and the product was stored. The chemical analyzes of the prepared feeds were made. Protein, fat, ash, yeast, mold and Mesophilic Aerobic Bacteria values of the feeds were compared. Results of live weight gain and economic analysis obtained from the study; It has been shown that semi-aged feeds made with Van fish and trout internals can be used in feeding Rainbow trout and will reduce feed costs.

Anormal Gonadlı Van Balığının (Alburnus tarichi Güldenstädt, 1814) Bazı Dokularında Oksidatif Hasarın Belirlenmesi

Van Lake is the largest soda lake of Turkey and there is only one fish species (Van Fish, Alburnus tarichi Güldenstädt, 1814) that can survive under the extreme conditions of the lake. There are several researches carried out on the issue of lake contamination recently. Gonad abnormalities observed among these fish are one of the most important issues. In the present study, the malondialdehyde (MDA) and reduced glutathione (GSH), and the antioxidant enzyme levels such as superoxide dismutase (SOD) and glutathione peroxidase (GSH – Px) are compared in gills, livers, and gonads of the Van Fish (the ovary abnormality of which are determined in Van Lake) and the normal fish. Compared to the normal fish, the MDA levels of the abnormal fish show a considerable amount of increase in all of the tissues (P<0.05). There is a significant decrease in the SOD activity in the gill and ovary tissue and in the GSH – Px activity in the livers of the abnormal individuals compared to the normal individuals (P<0.05). As for the GSH levels, there is no difference found when the normal and abnormal tissues are compared. Consequently, when the fish with ovary abnormalities which are thought to be exposed to contaminators in a certain time period is compared to the normal fish, it can be said that their antioxidant defense systems are affected.

Development of osmoregulatory tissues in the Lake van fish (Alburnus tarichi) during larval development.

Lake Van is one of the largest alkaline lakes worldwide and Lake Van Fish (Alburnus tarichi Güldenstädt, 1814) is the only vertebrate species inhabiting it. Lake Van Fish is an anadromous species that migrates to the streams (salinity 0.02%, pH8.42) flowing into Lake Van (salinity 0.22%, pH9.8) during the spawning period (April-July). Following spawning, fish return to Lake Van while larvae remain in fresh water. This study examined the development of osmoregulatory organs and the distribution of ionocytes in Lake Van Fish larvae adapting to the highly alkaline water characterizing the lake. Ionocytes were marked immunohistochemically and observed in whole mounts with immunofluorescence staining using the Na+/K+ ATPase antibody. Ionocytes were first identified in the yolk sac membrane and skin, and then in the gills, digestive tract, and kidneys of larvae. The number of ionocytes on yolk sac membrane and skin decreased during larval development, indicating ionocytes on these tissues have a role in larvae osmoregulation. Larvae hatched from eggs in stream waters die when transferred to Lake Van water but survived in lake water diluted with deionized water. Thus, larvae need to go through certain alterations at the cellular and organ levels in order to adapt to the conditions of Lake Van water, indicating they do not enter this lake immediately after hatching.

Histological Organization of Thyroid and Interrenal Glands of the Pearl Mullet, Alburnus tarichi (Cypriniformes: Cyprinidae) from Lake Van Basin of Turkey

In this study, histological organizations of the thyroid and interrenal glands were first investigated in pearl mullet (Alburnus tarichi Guldenstadt, 1814), living in the Lake Van basin in Turkey. The serial sections taken from the subpharyngeal region and kidney were stained with hematoxylin and eosin, Mallory’s trichrome and periodic acid-Schiff/hematoxylin and then examined. The histological examinations showed that the thyroid gland of pearl mullet was unencapsulated and the thyroid follicles were scattered diffusely in the subpharyngeal region, located at the dorsal and lateral aspects of the ventral aorta. The follicles had colloid-filled lumens and were surrounded by simple follicular epithelial cells varying in shapes (squamose, cuboidal and cylindrical). Thyroid follicles were not found in any areas of the kidney. The interrenal and chromaffin tissue were observed to be associated with postcardinal vein with its branches in the head of the kidney. The interrenal cells having a deeply eosinophilic cytoplasm were determined adjacent to branches in the head kidney. The eosinophobic chromaffin cells were found to show two different distributions. They were observed as being intermingled with the interrenal cells and formed separate groups surrounded by a thin connective tissue in the walls of the major veins. Both interrenal and chromaffin cells displayed a negative periodic acid-Schiff reaction.