Vesna Poleksić

Liver, gills, and skin histopathology and heavy metal content of the Danube sterlet (Acipenser ruthenus Linnaeus, 1758).

The sterlet (Acipenser ruthenus L.) is a bottom-feeding fish species with a direct exposure to contaminants from water and sediments. Although heavy metal pollution is believed to be one of the main threats to the sterlet population in the Danube River basin, there is a lack of knowledge of the exact impact of heavy metals on their survival. In the present study, effects of heavy metal pollution on sterlet in the Danube basin were assessed as well as the utility of different sterlet organs and tissues as indicators of heavy metal contamination. The sterlet were sampled at three different sites in the Danube basin, in Hungary and Serbia, isolated from each other by dams. Heavy metal analysis included measurement of Cd, As, Pb, Cr, Hg, Cu, Ni, Fe, Mn, and Zn concentrations in sterlet gills, muscle, liver, and intestine, and histopathological analyses comprised assessment and scoring of the extent and intensity of alterations in skin, gills, and liver tissue. Analysis revealed a significant presence of sublethal histopathological changes that were most pronounced in the liver and skin and increased accumulation of heavy metals, with the highest concentrations in the liver. Canonical discriminant analysis showed significant differentiation among the three studied localities, suggesting that the heavy metal concentrations in sterlet populations were site specific. The present study concludes that the accumulation of heavy metals is a response to the presence of these pollutants in the environment, and, together with other pollutants, it affects the vital organs of natural sterlet populations.

Histological methods in the assessment of different feed effects on liver and intestine of fish.

This manuscript describes the importance of using histological methods to assess the effects of feed on the liver and intestine of fish. Due to the constantly increasing world production of fish and other aquatic organisms, it is necessary to replace fishmeal and fish oil in diets with less expensive raw materials of plant origin. Due to the increased fiber content, increased presence of carbohydrates, antinutritional factors, and inappropriate content of amino acids and new compounds can have negative effects on the digestive system of fish and therefore on fitness, health and production characteristics of cultivated fish. The liver and intestines are the most important organs for the digestion and absorption of nutrients from feed. Therefore, monitoring histological structure of fish liver and intestine is the method of choice in assessing the effects of nutrient mixtures that use raw materials of plant origin. For both of these vital organs the normal histological structure and the most important results obtained by research are discussed. This paper presents a critical review of the histological methods used in research on feed effects. Results related to the negative effects of raw soy-based feed on the occurrence of enteritis in carnivorous fish species are discussed. The results point out that use of modern approach in fish pathology such as improved histochemical, stereological, scoring, and other analytical methods could be a beneficial approach in an accurate assessment of new feed effects on fish.

Histopathological indicators: a useful fish health monitoring tool in common carp (Cyprinus carpio Linnaeus, 1758) culture

In order to evaluate the relationship between water quality in ponds and indices of histopathological changes occurring in the vital organs of the common carp (Cyprinus carpio L., 1758), two six-month field experiments were carried out using two different water supplies: from the nearby stream and a tube well. The fish were fed supplemental feed: raw cereals, pelleted and extruded compound feed. Histopathological analysis, alteration frequencies, and semi-quantitative scoring of the changes were used to assess the health status of the fish. Ponds supplied by stream water were characterized by higher water hardness, dissolved oxygen and pH values, while those supplied by the tube well had higher electroconductivity, total ammonium and orthophosphates content. Fish survival rate and habitat suitability index were lower in ponds supplied by stream water, while the weight gain did not differ between the two water supplies. The use of stream water resulted in a higher level of histopathological changes in gills and liver. Among the water quality parameters, pH level had the strongest influence on fish. Differences in water supply produced greater influence on the level of histopathological changes than the type of feed applied. Gills were the most sensitive organ, while the kidney was the least responsive.

Use of histopathology and elemental accumulation in different organs of two benthophagous fish species as indicators of river pollution.

The bottom feeding fish species have a good potential to be used for assessments of pollution, as they are under pressure from pollutants from both water and sediments. In this study, the level of similarity of histopathological responses to pollution in gills and liver between barbel (Barbus barbus) and sterlet (Acipenser ruthenus) from the Danube River was assessed, and compared with elemental concentrations in their gills, liver, and muscle. Results indicate that the detected metal concentrations were likely cause of different tissue responses in gills and liver of the two investigated fish species. Statistical analysis indicated a clear differentiation of the two species based on elemental concentrations and the level of histopathological changes in gills and liver. Metal concentrations exceeded maximum acceptable concentrations in a number of analyzed specimens, which indicates the importance of this type of monitoring. Results indicate that barbel is a better indicator for specific, rather narrow sites, whereas sterlet is a better indicator of larger (longer) water current segments. Obtained information could be of importance for both scientists and fishery and water management authorities working on the development of water monitoring programs.

Toxic effects of 2,4‐D herbicide on fish

Abstract Acute and subacute 2,4‐D toxicity to carp (Cyprinus carpio L.) were investigated. Acute toxicity (LC‐ 50) was investigated in semi‐static test during a 24, 48, and 96‐ hours exposition. Subacute toxicity was investigated by exposing fish to different 2,4,‐D concentrations (150, 200, and 250 mg/L) for 14 days. Biochemical and morphological changes in certain organs and tissues were investigated. LC‐ 50 values at 24 hours exposure was 310.0 mg/L, 295.0 mg/L for 48 hours, and 270.0 mg/L for 96 hours exposure. Subacute toxicity tests show that 2,4‐D induce changes in the enzyme activities (AP, GOT, GPT) and morphological changes in the gills, liver and kidneys, but changes are of limited biological importance.

The impact of multiple stressors on the biomarkers response in gills and liver of freshwater breams during different seasons

Biomarkers attract increasing attention in environmental studies, as a tool for detection of exposure and effects of pollution, from both natural and anthropogenic sources. This study aims to assess the impact of multiple stressors during distinctive seasons, covering also extreme hydrological events (extensive flooding in the mid May 2014), on different levels of biological organization in the liver and gills of three closely related freshwater breams. Our previous study on DNA damage in blood cells of these specimens showed increased DNA damage in June 2014, one month after the flooding event. As a continuation of that research, the present study was conducted. As a biomarker of exposure DNA damage was measured by applying the alkaline comet assay, while histopathological alterations were monitored as a biomarker of effect. Additionally, concentrations of metals and metalloids in gills, liver and muscle were assessed. Sampling of fish tissues was performed in 2014, during winter (January and February), spring (March and early June) and summer (late June, July and August). Significant seasonal difference in DNA damage was observed for both tissues. During spring and summer the level of DNA damage in gills was significantly higher when compared to the liver. Histopathological analyses showed higher frequency of alterations in gills during spring, and in liver during summer, but without a significant seasonal difference. Gills had the highest concentration of metals and metalloids during the spring and summer, and liver during winter. Muscle was the least affected tissue during all three seasons. This study highlighted the importance of the multiple biomarker approach and the use of different fish tissues in assessment of surface water pollution.

Effect of supplemental feeds on liver and intestine of common carp (Cyprinus carpio) in semi-intensive rearing system: histological implications

Abstract In order to determine effects of different types of supplementary feed on the histology of liver and intestine of carp reared in a semi-intensive system, a six-month field experiment was carried out. Carp yearlings were fed different supplemental feeds: cereals (CF), pelleted (PF) and extruded (EF) compound feeds. Hepatocytes histomorphometry revealed larger nuclear profile area of EF-fed fish compared to other two groups, while cytoplasmic profile area of the hepatocytes decreased in following sequence: CF > EF > PF. Lower nucleus/cytoplasm ratio was found in CF-fed fish compared to EF-and PF-fed fish. Striking, season dependant finding, was that regardless of the type of supplementary feed the number of lipid droplets in hepatocytes was highest in July and August, lipid depletion occurred in September and October, while glycogen was present in hepatocytes throughout the study. No size difference between groups was observed for enterocytes height, absorptive surface height and number of goblet cells in tunica mucosa, but a strong relationship between length of intestinal folds and fish weight was found. Histological analysis revealed a difference among three types of supplemental feed used in this field study: extruded or steam-pelleted feed induced better nutritional status of common carp in comparison to cereals. At the same time, histomorphometry indicated some differences between EF-and PF-fed fish.

Effects of mine tailing and mixed contamination on metals, trace elements accumulation and histopathology of the chub ( Squalius cephalus ) tissues: Evidence from three differently contaminated sites in Serbia

Chub (Squalius cephalus) specimens were collected in Korenita River seven months after spillover from the waste water of antimony mine tailing pond and compared with chub living in Kruščica reservoir (intended for water supply) and Međuvršje reservoir (influenced by intense emission of industrial, urban and rural wastewater). Concentrations of 15 elements (Al, As, B, Ba, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Si, Sr, Zn) were determined in muscle, liver and gills of chub by inductively coupled plasma optical spectrometry (ICP-OES) and histopathological alterations in liver and gills were assessed. Chub specimens from Korenita River had higher concentrations of As, Ba and Pb in all three investigated tissues as well as higher total histopathological index values than chub from reservoirs. Specimens from Međuvršje reservoir were characterized by higher values for concentrations of Cu and Si in muscle tissue and higher values for regressive histopathological alterations in gills. Individuals of chub from Kruščica reservoir had the highest concentrations of Fe in liver, Hg in muscle and Sr and Zn in muscle while gills had the lowest value of total histopathological index. The results from the present study showed higher level of histopathological alterations as a result of mine tailing accident. As a result of mixed contamination on the Međuvršje site, histopathological index values of gills were in line with the index value from Korenita River. Increased values for Fe and Sr in chub tissue from Kruščica reservoir could be explained by geological structure of the site which is characterized by magmatic rock rich in Cu, Fe and Ni as well as dominant carbonate sediment complex of marine origin with increased level of Sr.