Biochemical and toxicopathic biomarkers assessed in smallmouth bass recovered from a polychlorinated biphenyl-contaminated river

Smallmouth bass (Micropterus dolomieu) were collected to quantify the nature and prevalence of biomarker responses, including biochemical indices, toxicopathic lesions and general health indices, among fish collected from polychlorinated biphenyl (PCB)-contaminated and nearby uncontaminated reaches of the Kalamazoo River, Michigan, USA. Blood and tissue samples (gill, liver, spleen, head kidney, trunk kidney, thyroid and gonads) were collected and preserved at necropsy for biochemical and histological analyses. The body condition factor and liver somatic index were significantly lower in fish collected from the downstream, contaminated site. Plasma vitellogenin was not detected in male fish collected from either site. Liver ethoxyresorufin-O-deethylase activity and liver and spleen superoxide dismutase activity were significantly depressed in fish collected from the downstream site. Significant toxicopathic lesions such as glycogen depletion, enhanced macrophage aggregates, hepatic foci of cellular alteration (i.e. preneoplastic lesions) and neoplasia were also detected in the liver of fish collected from the downstream site. This study indicates that many of the biochemical and histopathological biomarker responses were associated with liver and body tissue PCB concentrations. Taken together, the biomarkers of exposure and effect strongly suggest that fish within the downstream site are adversely affected by PCBs and other chemical stressors.     

Effects of some pesticides on the vital organs of juvenile rainbow trout (Oncorhynchus mykiss)

Gill, trunk kidney, spleen, and liver of rainbow trout (Oncorhynchus mykiss) were examined after exposure to different sublethal concentrations of carbosulfan (25, 50 and 200 μg L⁻¹), propineb (3, 6 and 24 mg L⁻¹), and benomyl (2, 5 and 20 mg L⁻¹) for 14 days. Lesions were observed in gill, trunk kidney, spleen, and liver of rainbow trout exposed to either concentration of pesticides. The most important lesions were determined in the highest concentrations of pesticides. Lamellar fusion, lamellar hyperplasia, epithelial lifting, vacuolization of epithelial tissue, epithelial necrosis, hypertrophy and sloughing of epithelium were observed on fish exposed to carbosulfan, propineb and benomyl. Fish had cell necrosis, degeneration and oedemas in liver, trunk kidney and spleen. None of these lesions were seen in control fish.     

Seasonal changes in morphometric and biochemical endpoints in northern pike (Esox lucius), burbot (Lota lota) and slimy sculpin (Cottus cognatus)

1. Morphometric and biochemical endpoints were used to assess growth and energetics in young‐of‐the‐year (YOY) northern pike (Esox lucius), YOY burbot (Lota lota) and slimy sculpin (Cottus cognatus) collected upstream and downstream of a uranium mining operation in northern Saskatchewan, Canada. 2. Water sampled at sites downstream of uranium mining and milling effluent discharge was higher in certain trace metals (arsenic, molybdenum), ions (conductivity, hardness, ammonia and sulphate) and total dissolved solids compared with ecologically similar reference sites. 3. Overwinter survival potential was estimated by determining total body lipids and total body triglycerides in fishes collected in late autumn and early spring. Two‐factor anova indicated no site or season differences in total body lipids or triglycerides in YOY northern pike. In spring, burbot collected from the exposure site were higher in total body lipids and triglycerides compared with the previous autumn and compared with fish collected from the reference site. Slimy sculpin from both exposure and reference sites had lower total body lipid and triglyceride content in spring compared with the previous autumn. In addition, sculpin from the exposure site had lower total body lipids and triglycerides in spring compared with fish collected from the reference site. 4. Biochemical estimates of fish growth (muscle RNA/DNA ratio and muscle protein concentration) exhibited clear seasonal differences, with consistent decreases in this endpoint for all fish species in spring relative to the previous autumn at both exposure and reference sites. 5. Results for total body lipids and triglycerides were inconclusive and inconsistent between species and exposure scenarios. However, all species exhibited a similar seasonal decrease in muscle protein and to a lesser extent, muscle RNA/DNA ratio.     

Establishing the Causal Relationship between Polycyclic Aromatic Hydrocarbon (PAH) Exposure and Hepatic Neoplasms and Neoplasia-Related Liver Lesions in English Sole (Pleuronectes vetulus)

For almost 25 years our laboratory has studied the impact of PAHs and related industrial contaminants on benthic fish, following an interdisciplinary approach involving chemical exposure assessment linked to synoptic detection of various effects at several levels of biological organization. These data demonstrate a cause-and-effect relationship between neoplastic and neoplasia-related liver lesions in English sole, and exposure to PAHs, and to a lesser degree, chlorinated hydrocarbons such as PCBs. In statistical analyses of data from multiple field studies conducted since 1978, exposure to PAHs measured in various compartments has consistently been identified as a highly significant, major risk factor for neoplasms and related lesions in this species, with PCB exposure shown to be a significant, but less consistent and less strong risk factor for these lesions. A cause-and-effect relationship between PAHs and toxicopathic liver lesions in this species is further supported by the experimental induction of toxicopathic lesions identical to those observed in field-collected fish, in sole exposed in the laboratory to model carcinogenic PAHs such as BaP or to PAH-rich extracts of sediments from Eagle Harbor, a severely PAH-contaminated site in Puget Sound. More recent field studies have identified significant associations between hepatic cytochrome P4501A (CYP1A) induction and xenobiotic-DNA adduct formation, and hepatic lesion prevalences in wild subadult English sole. Field studies in Eagle Harbor subsequent to capping of the most PAH-contaminated region of this harbor with clean dredge spoils have shown a decline in exposure to PAHs as assessed by biliary fluorescent aromatic compounds (FACs) and hepatic xenobiotic-DNA adducts. This decline in PAH exposure has been accompanied by a dramatic decline in risk of occurrence of toxicopathic hepatic lesions in English sole from Eagle Harbor. Further, laboratory studies have induced lesions in English sole by injections of extracts from PAHcontaminated sediments. Overall, these findings relating to exposure to PAHs and chlorinated hydrocarbons and the occurrence of hepatic neoplasms and neoplasiarelated lesions in English sole fulfill the classic criteria for causality in epizootiological or ecological risk assessment studies, including: (1) strength of association, (2) consistency of association, (3) specificity of association, (4) toxicological and biological plausibility, (5) temporal sequence/timing (i.e., exposure precedes disease, effect decreases when the cause is decreased or removed), (6) dose-response or biological gradient, and (7) supportive experimental evidence.     

Field Studies Using European Starlings to Establish Causality between PCB Exposure and Reproductive Effects

Accumulation and effects of polychlorinated biphenyls (PCBs) in avian species were evaluated at a Superfund site located at Crab Orchard National Wildlife Refuge, Illinois, and seven criteria were used to assess whether there was a causal relationship between PCB exposure and observed reproductive effects. European starlings (Sturnus vulgaris) were monitored at nest boxes constructed at each of two exposed and two reference sites. During the breeding season, starling productivity (number of chicks produced per nest) and adult nest attentiveness behavior (provisioning behavior) were monitored. At 15 days post-hatch, chicks were collected for contaminant and biomarker analyses. Chicks were necropsied, ethoxyresorufin-O-deethylase (EROD) activity measured in liver tissue, and PCB (Aroclor 1254) and 34 chlorinated biphenyl (CB) congener concentrations measured in carcasses using gas chromatography. PCB and CB concentrations also were measured in eggs that failed to hatch. Mean Aroclor 1254 and quantified CB concentrations were greater (P<0.001) in eggs that failed to hatch and 15-day-old chicks collected from PCB sites compared to those collected from reference sites. EROD activity was greater (P=0.005) in 15-day-old chicks collected from PCB sites and was correlated with carcass PCB concentrations. Reduced adult nest attentiveness behavior and decreased chick survival were observed at PCB sites. Six of the seven causal criteria evaluated provided evidence that observed reproductive effects resulted from exposure to PCBs. Using this weight-of-evidence approach provided a means for establishing the likely cause of effects and thus provided managers with information needed in decision-making processes.     

Nevirapine in African surface waters induces liver histopathology in Oreochromis mossambicus: A laboratory exposure study

Nevirapine (NVP) is one of the HIV antiretrovirals detected recurrently in African surface waters. Liver side effects in humans were reported. Because the effects on fish are unknown, this study investigated the potential effects of NVP on selected biometric indices and liver histology of Oreochromis mossambicus (Mozambique tilapia) in a chronic exposure. Adult fish were exposed to two concentrations of NVP (1.48 and 3.74 µg l^?1) in a static renewal system under controlled conditions for 30 days. At the end of the exposure, each fish was weighed, the total length recorded, and blood collected from the caudal vein. A standard necropsy was done, selected organs were sampled and weighed, and biometric indices calculated. Liver tissue was processed for histological assessment. Results showed significant differences (p < 0.001) in hepatosomatic indices between the exposed fish and the control, as well as in spleen indices of the fish exposed to the higher NVP concentration (p = 0.008). Liver tissue of exposed fish showed significant histological changes (p < 0.001), including hepatocyte apoptosis, vacuolation, and a mild fibrosis around some of the veins and bile ducts. Nevirapine in African surface waters could have long-term negative effects on the health of fish.     

Health and chemical burdens of fish species from polluted and hyper-eutrophic freshwater ecosystems in South Africa

Three aquatic ecosystems in South Africa, the Hartbeespoort, Klipvoor and Bospoort Dams, are classified as hyper-eutrophic, because of high nutrient loads and chemical pollution. Water and two fish species, Clarias gariepinus and Cyprinus carpio, were collected from these dams to assess the impact of eutrophication and chemical pollutants on their health status. Water and muscle samples were analysed for organic and inorganic chemicals. Condition factor was determined and a necropsy performed to note any macroscopic abnormalities. A histology-based fish health assessment was done on the liver, kidney, gills and gonads. A number of fish from the three dams exhibited livers with fatty change and focal discoloration, skin lesions and parasites within the visceral cavity. The prevalence and severity of histopathology in the liver resulted in higher liver index values than the index values for kidneys and gills. Aluminium, silicon and chromium were detected in the water and muscle tissue. The DDT metabolite p,p’-DDE was present in both species, as well as in fish from the reference site, Marico-Bosveld Dam. Only C. gariepinus from Hartbeespoort Dam had p,p’-DDD levels higher than 5 µg g^?1 per edible portion. Water from hyper-eutrophic dams adversely affects the health of freshwater fish.     

Remarkable Shrimps: Adaptations and Natural History of the Carideans

A histology-based fish health assessment protocol was used in 2009–2010 to assess the health status of Clarias gariepinus from Pongolapoort Dam, South Africa. Nineteen fish were collected by angling. The histology of liver, kidney, gills and testes or ovaries was semi-quantitatively assessed and compared to that of fish from a reference site in the Okavango Panhandle, Botswana. Necropsy observations indicated that C. gariepinus from Pongolapoort Dam were in poorer condition than those from the Okavango Panhandle. Microscopic analysis showed that the mean liver index (I _L), gill index (I _G), testis index (I _T) and ovary index (I _O) were higher, although not statistically significant, in fish from the Okavango Panhandle. The difference in the results between the necropsy-based observations and semi-quantitative histological results was due to the external abnormalities detected in the necropsy analysis, including parasites and skin lesions, not expressing as changes to cell and tissue structure. According to the parameters measured, C. gariepinus from Pongolapoort Dam were in a healthy state.     

Interdemic variation in haematocrit and lactate dehydrogenase in the African cyprinid Barbus neumayeri

This study evaluated whether the African cyprinid Barbus neumayeri from Rwembaita Swamp (low‐oxygen) and Njuguta River (high‐oxygen) in the Kibale National Park, Uganda differed in traits related to aerobic and anaerobic metabolic potential. Haematocrit was measured as an index of blood oxygen‐carrying capacity, and tissue activities and isozyme composition of lactate dehydrogenase (LDH) were measured as indices of tissue anaerobic capacity. To address whether site‐dependent differences were acute responses v . longer‐term adjustments to environmental conditions, these variables were measured in fish sampled shortly after collection and after laboratory maintenance under well‐aerated conditions. In fish sampled in the field, those from the low‐oxygen site had significantly higher haematocrit, but this difference disappeared after long‐term laboratory maintenance. In contrast, fish from the low‐oxygen site had higher liver LDH activities than fish from the high‐oxygen site, and this difference persisted during laboratory maintenance. Polymorphism was detected at both the LDH‐A and LDH‐B loci, and genotype frequencies for LDH‐B differed significantly between collection sites. These results demonstrate physiological, biochemical and genetic differences in B. neumayeri from habitats differing in dissolved oxygen availability and suggest both acute and long‐term responses to local environmental conditions.     

The interactive effect of elevated temperature on deltamethrin-induced biochemical stress responses in Channa punctata Bloch

There are reports showing interactive effect of environmental factors with the toxic outcome of chemicals. We studied the interactive effect of elevated temperature as an abiotic stressor on deltamethrin-induced biochemical stress responses in a freshwater fish, Channa punctata Bloch. Heat stress (∼12 °C above ambient temperature for 3 h) and pesticide exposure (deltamethrin 0.75 ppb for 48 h) showed significant induction of heat shock protein-70 (HSP70) in liver, kidney and gills of fishes. Elevated temperature when followed by deltamethrin exposure showed synergistic effect showing a high level of HSP70 in liver and gills whereas response in the kidney was opposite. On the contrary, when deltamethrin exposure followed the heat stress, no significant difference was observed. Protein carbonylation was found to be more pronounced in heat-stressed group compared with control fish group. A significant increase in lipid peroxidation (LPO) was observed in different tissues of fish exposed to either of the stressors. In the kidney of fish exposed to heat stress followed by deltamethrin, LPO was relatively lower as compared to other treatments. Thiols content such as reduced glutathione (GSH), total thiols (T-SH), non-protein thiols (NP-SH) and protein thiols (P-SH) showed no consistent pattern in different tissues. In deltamethrin-exposed group that was subsequently exposed to heat stress, the GSH content was higher in liver and lower in both kidney and gills when compared with other groups. Alteration in the activities of antioxidant enzymes such as catalase (CAT), glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx) was also observed when fish were exposed to heat stress and/or deltamethrin. Our study demonstrated that heat stress modulated biochemical stress responses in fish showing a tissue specific pattern. This implies that fish has the capacity to elicit differential response to exposure to abiotic stressors in order to reduce the systemic magnitude of stress which may otherwise lead to severe dysfunction of vital tissues.     

Isolation and characterization of Pseudomonas sp. KUMS3 from Asian sea bass (Lates calcarifer) with fin rot

Bacteria were isolated from Asian Sea bass, Lates calcarifer kept in a farm, on the South-east coast of India. During an outbreak of fin rot, the affected fish had hemorrhages at the base of fins, mouth and skin muscles and faded pigments. Pure colonies were isolated on NA and ZMA from internal organs of the fish and the bacterial morphology was identified as gram-negative rod-shaped bacteria. Based on different biochemical tests and sequence of 16S rDNA, the causative bacteria were identified as Pseudomonas sp. KUMS3. Bacterial cells were isolated from liver and kidney of all artificially infected moribund fish and confirmed as Pseudomonas sp. KUMS3 by morphological and biochemical characteristics. During the experimental infection, the first incidence of dead fish was observed on 2nd day after exposure to Pseudomonas sp. KUMS3 and no fish died after 12 days post exposure and the cumulative percent of mortality was 70. Histological lesions were observed in the spleen, liver and kidney of the infected fish. Pseudomonas sp. KUMS3 could be considered as an opportunistic pathogen, which can survive on the fish surface or in water or in the gut and may cause disease when unfavorable conditions develop.     

Functional dissimilarity of melanomacrophage centres in the liver and spleen from females of the teleost fish <Emphasis Type="Italic">Prochilodus argenteus</Emphasis>

Melanomacrophage centres (MMCs) are formed by macrophage aggregates containing pigments such as hemosiderin, melanin and lipofuscin. MMCs are found in animals such as reptiles, amphibians and, mainly, fishes, in organs such as the kidney, spleen, thymus and liver. In teleost fish, several functions have been attributed to MMCs, including the capture and storage of cations, the phagocytosis of cellular debris and immunological reactions. As the use of MMCs has been suggested as a tool for the assessment of environmental impacts, our aim has been to describe the various metabolic processes performed by MMCs in diverse organs (liver and spleen) by using the teleost Prochilodus argenteus as an animal model. MMCs from the liver and spleen were assessed by histochemistry, transmission electron microscopy, scanning electron microscopy, X-ray microanalysis techniques and biochemical assay for N-acetylglucosaminidase activity. The data showed metabolic differences in MMCs between the liver and spleen of P. argenteus in their morphometric characteristics and biochemical and elemental composition. The implications of these findings are discussed, focusing on their role in organ metabolism.     

The histophysiology of antibody-forming sites in the marine toad

Summary The normal histologioal features of the lymphatic organs — pericardial nodes, jugular bodies, spleen, and kidney — of the marine toad, Bufo marinus, are decribed. The lymph nodes and spleen of the marine toad lack the compartmental organization of corresponding mammalian organs and contain relatively less internal connective tissue. The cellular stroma composed of reticulum cells and fixed macrophages plays a more important role in maintaining structural organization than do the connective tissue.Changes in the cellular composition of the lymphatic parenchyma were observed in animals immunized with bovine serum albumin suspended in Freund's complete adjuvant. In addition to an increase in the number of lymphocytes and the presence of lymphoid hemocytoblasts, cells occurred which possessed many of the morphological characteristics of mammalian plasma cells. These plasma cells, which exhibited positive fluorescent antibody reaction, were more abundant in the kidney than in the lymph nodes or spleen of an immunized animal.Granulomas developed at the site (gastrocnemius muscle) of injection of antigen in complete adjuvant, and similar cystic lesions arose in the kidney. Apparently, the antigen-adjuvant mixture found its way from the site of injection (gastrocnemius muscle) into the kidney, probably via the renal portal system, and established lesions in the kidney. Appreciable numbers of antibody-forming cells, or plasma cells, were found in the muscle granulomas and in the kidney lesions.The lymphoid tissue of the kidney is considered the principal site of antibody formation in the marine toad, Bufo marinus.This investigation was supported by grants HD-2614-1 and GM-11782 from the United States Public Health Service administered by Dr. Ronald R. Cowden and Dr. E. Peter Volpe, respectively.     

Histopathological biomarkers and genotoxicity in gill and liver tissues of Nile tilapia Oreochromis niloticus from a polluted part of the Nile River,Egypt

Fish health is affected by water pollution. Oreochromis niloticus collected during summer 2014 from El-Serw, a polluted site on the Nile River, were compared with fish from a reference site, El-Zamalek. Histopathological changes were detected in gill and liver tissue samples using light and electron microscopy. In addition, the degree of DNA damage was measured using the comet assay. To indicate the severity of water pollution at the two sites, physico-chemical properties and heavy metal concentrations were investigated. Gill damage, including lamellar cell hyperplasia and aneurysm, was observed in the fish samples from the polluted site. The livers of fish from the polluted area showed necrosis and an increase in melanomacrophage centres. Histochemical results confirmed a marked rise of gill mucopolysaccharides and a reduction of carbohydrate stored in hepatocytes. Electron microscopy revealed clear alterations in gill and liver tissue of fish from the polluted site. The comet assay showed highly significant DNA damage in tilapia collected from the polluted site, compared to those from the reference site. Histopathological biomarkers and the comet assay may therefore be sensitive indicators of exposure to mixtures of aquatic pollutants in Nile tilapia.     

D-Amino acid oxidase in various fishes*

A systematic procedure is provided for determination of D-amino acid oxidase (EC 1.4.3.3) activity in fish tissues. Activity was surveyed in crude liver homogenates of twenty-three species of fish. Activity ranged from 0-047 to 3-12 units (ρmol product produced per min) per gram wet weight of tissue. Activity was highest in liver, with significant levels occurring in kidney and pyloric caecae. It was below the limit of detection in brain, heart, spleen, testis, gill filaments, and muscle. Some properties of the enzyme from sockeye salmon, Oncorhynchus nerka (Walbaum), were studied. Knowledge of occurrence and levels of D-amino acid oxidase in fish may be useful to fish aquaculturists contemplating supplementation of feed with racemic mixtures of amino acids.     

Variation in the summer diet of age-0 brown trout in a regulated mountain stream

We describe the age-0 brown trout (Salmo trutta) collected from a mountain stream downstream from a reservoir in June, July, and August 1990. Sample sites were 1, 12, and 20 km downstream from the reservoir at elevations of 2840, 2600, and 2230 m above mean sea level. The mean length of fish in the samples increased with downstream progression and from June through August. Zooplankton predominated the diet of fish collected in June from the most upstream sampling site, while Diptera larvae predominated the diet of fish from more downstream sampling sites in June. In July and August, the diet of fish from the upstream site was predominated by Diptera larvae, while the diet of fish from the other two sites was predominated by Ephemeroptera nymphs. Variation in diet among sampling sites and sampling months was related not only to increasing fish sizes over the summer but also to the availability of zooplankton flushed from the reservoir.     

Expression of 8-OHdG in Zosterisessor ophiocephalus from the Venetian lagoon,Italy

The aim of the present work was to evaluate the expression of 8-OHdG (8-hydroxy-2''-deoxyguanosine) in the benthic fish Zosterisessor ophiocephalus collected in two differently polluted sites of the Venetian lagoon (Porto Marghera and Caroman). We compared our data on 8-OHdG with those of CYP1A (Cytochrome P450, family 1, subfamily A, polypeptide 1), which is a well known biomarker for detoxification of contaminants. Immunohistochemistry with an antibody to 8-OHdG showed immunopositivity in nuclei of hepatocytes as well as in melanomacrophage centres of spleen and kidney, whereas an anti-CYP1A antibody exhibited positive immunostaining in the liver, kidney and ovary. The liver of males showed higher expression of both proteins than females. In animals from Porto Marghera site, the enzymatic assay for 8-OHdG exhibited higher levels in liver of males than in females. Western Blot analysis using the antibody anti-CYP1A recognized the presence of a band of about 60 kDa in the liver of males and females. Males exhibited a strong band, whereas in females the band showed a lower intensity. By using Real-Time PCR, the mRNA expression of CYP1A did not show any differences between males and females from each site, but it was at borderline significance level. Comparing the two sites, mRNA expression of CYP1A was significantly higher in the liver of both males and females from Porto Marghera than that of Caroman. The present data suggest that pollutants are bio-available as demonstrated by our biomarker analyses and may have a harmful effect on aquatic organisms such as Z. ophiocephalus. We report that the highest levels of hepatic 8-OHdG and CYP1A expression were detected in males, showing clear gender specificity.Key words: OHdG, CYP1A, fish, immunohistochemistry, molecular biology.     

饲料中维生素D3的添加水平对黄颡鱼幼鱼生长和Toll样受体TLR18、TLR19和TLR21的影响

为了探讨饲料中维生素D₃添加水平对黄颡鱼(Pelteobagrus fulvidraco)生长和Toll样受体的影响, 研究设计了5个不同浓度梯度的维生素D₃饲料(1120、2260、3950、8030和16600 IU/kg), 对体重为(5.0±0.2) g的黄颡鱼进行了为期12周的生长实验, 并在生长实验结束后进行鮰爱德华氏菌(Edwardsiella ictaluri)攻毒72h。于攻毒前(0)和攻毒后(72h)采样, 每个饲料组分别取6尾鱼的脾脏、头肾、肝脏和前肠四个组织, 检测不同浓度维生素D₃处理对攻毒前和攻毒后TLR18、TLR19和TLR21基因表达量的影响。同时另取6条新鲜黄颡鱼的肌肉、头肾、肾脏、皮肤、脑、鳃、脾脏、胃上皮、小肠和肝脏, 检测TLR18、TLR19和TLR21基因在黄颡鱼中的组织分布。结果表明: 不同的维生素D₃添加水平会显著影响黄颡鱼幼鱼的生长性能; TLR18、TLR19和TLR21基因在所检测的组织中均有表达, 但在脾脏中表达量最高; 饲料中不同维生素D₃含量在攻毒前后均会显著影响TLR18、TLR19和TLR21在头肾、脾脏、肝脏和前肠中的表达, 攻毒后基因的表达显著高于攻毒前; TLR18、TLR19和TLR21在不同组织中的表达和饲料中维生素D₃的浓度相关, 研究结果表明饲料中添加合适剂量的维生素D₃, 可以促进相关免疫基因的表达, 从而增强黄颡鱼对病原微生物的抵抗力。     

板齿鼠内脏器官的重量和含水量的季节变化

本文研究板齿鼠部分内脏器官的鲜重、相对重量（脏器指数）和含水量的季节变化.结果表明：板齿鼠肝、心、脾、肾脏指数存在季节性变化,同时肝脏指数存在年龄和性别差异;脾脏指数与体重呈正相关,肝、肾脏指数与体重呈负相关;脏器水分含量只有肝和心脏存在季节性差异,但不存在年龄和性别差异.说明季节因素对不同脏器指数影响不同.