Characteristics of Melanomacrophage Centers in the Liver and Spleen of the Roach Rutilus rutilus (Cypriniformes: Cyprinidae) in Lake Kotokel during the Haff Disease Outbreak

ISSN 0032�9452, Journal of Ichthyology, 2014, Vol. 54, No. 1, pp. 104–110. © Pleiades Publishing, Ltd., 2014.Original Russian Text © S.V. Pronina, M.D.�D. Batueva, N.M. Pronin, 2014, published in Voprosy Ikhtiologii, 2014, Vol. 54, No. 1, pp. 107–114.

104

Lake Kotokel is the largest lake in Cisbaikalia andhas an area of 69 km2 and an average depth of 3.5 m. Itis located 2 km from the eastern shore of Lake Baikal(50°00′–50°50′ N, 108°05′–108°15′ E) and flows intoit through the Istok channel–Kotochik River–TurkaRiver system. According to the results of comprehen�sive research conducted in 1985–1986, this lake wasdescribed as a bicarbonate�calcium low�salinity waterbody of an eutrophic type with a roach and bass fishcommunity. Fish production in this lake in that periodreached 500 tons/year (Bioproduktivnost’ …, 1988).Twenty years ago, catches in this water body began todecrease, and today it is almost withdrawn from thefell fishery fund of the Buryat Republic. Exposure tonatural and anthropogenic factors, including high�level recreational pressure and unintended invasionand rapid development of the alien species Elodeacanadensis with its subsequent dying off, caused signif�icant changes in the structure of the biota of the lake(Pronin et al., 2007), as a result of which eutrophica�tion of the water body shifted towards dystrophication.In spring 2008, deaths of domestic cats that were fedwith fish and wild fish�eating birds were registerednear Lake Kotokel. In May and August 2008, massdeath of fish (roach Rutilus rutilus, perch Percae fluvi�atilis, and bream Abramis brama) was observed, whichcontinued sporadically in June–July 2009 (Pronin

1

2 2 3

et al., 2009). In July 2008, an outbreak of a diseasecaused by eating fish from Lake Kotokel by the localpopulation was registered, which was diagnosed asHaff disease (synonyms: Iuksov, Sartlan, Nariman,and Haff–Sartlan disease) or alimentary toxic parox�ysmal myoglobinuria (ATPM) (Kachin and Komar�ova, 2009). This disease was first registered in 1924 inthe inhabitants of the coast of the Frisches–Haff Bay(hence the name, Haff disease) in East Prussia (nowKaliningrad oblast). To date, a large number of out�breaks of this disease in different regions of Russia andabroad have been described; however, this is the firstcase for East Siberia and Baikal. Despite the long his�tory of studying APTM, the etiology of this diseaseremains obscure. Nevertheless, in each case, its occur�rence was associated with eating fish, and the toxicsource of it is heat�resistant and fat�soluble. Data onthe nature of the toxic substance accumulating in fishin APTM are ambiguous, which is probably due to thedifferent sources of its formation in different waterbodies. In view of this, a comprehensive research ofthe natural conditions and biota of Lake Kotokel wasperformed, including the ichthyoparasitological, his�topathological, and toxicological studies, whoseresults were described in part in a number of papers(Pronin et al., 2009, 2010; Pronina and Pronin, 2009;Bazova et al., 2010; Belykh et al., 2011).

Characteristics of Melanomacrophage Centers in the Liverand Spleen of the Roach Rutilus rutilus (Cypriniformes: Cyprinidae)

in Lake Kotokel during the Haff Disease OutbreakS. V. Proninaa, M.D.�D. Batuevab, and N. M. Proninb

a Buryat State University, Ulan�Ude, Buryat Republic, Russiab Institute of General and Experimental Biology, Siberian Branch, Russian Academy of Sciences,

Ulan�Ude, Buryat Republic, Russiae�mail: [email protected]

Received July 18, 2012

Abstract—Characteristics of morphology and number of melanomacrophage centers (MMCs) in the liverand spleen of the roach Rutilus rutilus and the amount of pigments in MMCs during the Haff disease outbreakand the death of fish in Lake Kotokel in relation to these parameters in the roach from Lake Baikal aredescribed. Pathological changes in the microvasculature and parenchyma in the liver of the roach from LakeKotokel were found. The area of melanomacrophage centers in the liver of the roach from this lake was sig�nificantly smaller, whereas the number and size of these centers in the spleen was significantly larger than inthe roaches from Lake Baikal. Among the pigments studied, the strongest response to the content of this toxinin the water body was shown by hemosiderin. An increase in its amount in the spleen MMCs testifies to anenhanced degradation of erythrocytes and iron release, which may be caused by the damage of cells of theerythrocyte lineage by the toxin.

DOI: 10.1134/S003294521401010X

Keywords: roach Rutilus rutilus, histopathology, melanomacrophage centers, spleen, liver, Haff disease, hepa�totoxin, microcystin

2 2

2 2

JOURNAL OF ICHTHYOLOGY Vol. 54 No. 1 2014

CHARACTERISTICS OF MELANOMACROPHAGE CENTERS 105

Melanomacrophage centers (MMCs) of thespleen, liver, and kidney as part of the defense systemof fish, amphibians, and reptiles are more and moreoften used as an object of micropathomorphologicaland toxicological studies (Mackmull and Michels,1932; Roberts, 1975; Wolke et al., 1985; Scalia et al.,1988; Pintucci et al., 1990; Wolke, 1992; Khan et al.,1994; Ahti Haaparanta et al., 1996; Couillard andHodson, 1996; Zuasti et al., 1998). The main func�tions of MMCs are the storage, destruction, anddetoxification of exogenous and endogenous materi�als, including parasites (Roberts, 1975; Wolke, 1992;Agius and Roberts, 2003).

The goal of this work was to perform a comparativestudy of MMCs in organs of one fish species fromLakes Baikal and Kotokel with contrasting ecologicalsituations.

MATERIALS AND METHODS

To study the histopathological changes in liver andspleen of roach, 16 individuals aged 6+ to 7+ wereused, which were caught June 5 and 6, 2009, in LakeKotokel. The liver and spleen of 15 roaches of thesame age caught June 8 and 9, 2009, in an environ�mentally safe water body (Chivyrkui Bay of LakeBaikal, 53°39′ N, 109°00′ ' E) served as a control.Chivyrkui Bay (area, 270 km2; length, 27 km; width,13 km) of Lake Baikal was taken as a control waterbody because it to the least extent experiences anthro�pogenic impact due to scarce residential populationon the adjacent territory, which belongs to the Trans�Baikal National Park, and due to the presence of aneutrophic area with an ichthyofauna similar to that ofLake Kotokel on its unique longitudinal transect withdepths ranging from 2 to 150 m (Pronin, 2000).

The organs were subjected to histological process�ing by the conventional methods used in pathohistol�ogy (Merkulov, 1969). After dehydration, fragments oforgans were embedded in paraffin. Paraffin blockswere cut into 4–6 μm thick sections with an Accu�CutSRM 200 microtome and stained with Ehrlich hema�toxylin—eosin, azure II—eosin, Mallory’s method,PAS reaction with an amylase control, 0.5% aqueoussolution of toluidine blue, and Berlin blue accordingto Pearls for differential identification of pigments(hemosiderin, melanin, and lipofuscin). Melanomac�rophage centers were counted per 1 mm2, the averagearea occupied by all MMC per 1 mm2 section was cal�culated, and the average content of pigments wasdetermined through an eyepiece disc with 100 pointsin 100 fields of view at a magnification of ×1000(Avtandilov, 1990).

Data were statistically processed using one�wayANOVA (Statistica and StatSoft (Russia) programpackages).

RESULTS

Liver (Fig. 1). In the roach from Lake Kotokel,unlike the fish from the control water body, significantpathological changes in the microvasculature andparenchyma were identified: mosaic perfusion of ves�sels (hyperemia and erythrostasis in the central veinsand sinusoids of some lobes and anemia in others);foci of perivenular edema of the parenchyma, diffusegranular and focal small�drop fatty degeneration; andsmall�focus (three to four cells) necrobiosis andnecrosis of hepatocytes (Fig. 1c). Staining accordingto Pearls showed the presence of small spherical darkblue granules of hemosiderin in the cytoplasm ofhepatocytes and stellate cells (macrophages) of sinu�soids, indicating the accumulation of iron in hepato�cytes (Fig. 1d). In one specimen (6.6%), histopatho�logical changes in the liver were most profound. Thebeam�type location of hepatocytes was disturbed,blood filling of vessels was weak, and many hepato�cytes had the signs of atrophy. Such cells had smallnuclei with eccentrically located nucleoli or weredevoid of them, with a coarse�grained dense cyto�plasm around the nucleus. The peripheral part of suchcells was light, with single basophilic granules. Theorgan contained foci of large�drop fatty degeneration.

The size of MMCs in the liver of the roach fromLake Kotokel and the control water body varied in therange from 86 to 2802 μm (table); the largest of themwere usually spherical. There was no distinct pattern inthe distribution of MMCs over the organ; however,some of them adjoined blood vessel walls. The leuko�cyte reaction around the MMC was missing, and theydirectly contacted with the parenchyma or blood ves�sel adventitial structures (Fig. 1a). Staining accordingto Mallory showed the presence of a thin collagenplate around some MMCs. It was found that mela�nomacrophage centers contained partly destructedhepatocytes and granules whose color varied fromlight yellow to brown (staining with hematoxylin–eosin) and from blue to violet (staining according toPearls). Melanomacrophage centers in the liver of theroach from Lake Kotokel, as compared to those in theliver of the control fish, did not contain melanin anddid not differ in the content of hemosiderin but weremuch more enriched in lipofuscin.

The liver of the roaches from the compared lakesdid not differ in the number of MMCs (table). How�ever, the area occupied by MMCs in the liver of theroach from Lake Kotokel was significantly smallerthan that of the roach from Lake Baikal due to thesmaller size of MMCs in the fish from Lake Kotokel(Figs. 1a, 1b).

Spleen. In the histological structure, this organhardly differed in the roach from the compared waterbodies; however, the number of MMCs in the spleen ofthe individuals from Lake Kotokel was 2.3 times greaterand they were much larger (49630 vs. 16450 μm2)(table). They are more intensively stained with hema�

106


PRONINA et al.

toxylin–eosin since they are richer in dark (almostblack) granules and simultaneously contain granulesof various shades of yellow and brown colors. Stainingaccording to Pearls revealed the presence of three pig�ments (hemosiderin, melanin, and lipofuscin) in thespleen MMCs of the roach from Lake Kotokel.Hemosiderin was detected in MMCs only in the spleenof the roach from Lake Kotokel. The spleen of fish fromthis lake contained significantly higher amounts of lipo�fuscin but less melanin (Table, Fig. 2a).

Melanomacrophage centers of medium and largesize were surrounded by a lymphocytic bank. Smalllymphocytes were found not only in the interstitiumbut also in blood vessels. In the last case, some MMCswere similar to the lymph nodules in the spleen ofmammals, except that there were pigment granules inplace of the reactive centers. On the border of somecenters and lymphocytic bank, a layer of flattened cellswith hyperchromatic nuclei was located, which wasexternally surrounded by a thin fibrillar plate that issometimes interrupted by inversion of leukocytes inthe interstitium (Fig. 2b). The flattened cells resem�bled endothelial cells. Probably, these MMCs formedin the lumen of sinus capillaries. Small MMCs directlycontacted with the hematopoietic tissue, because theywere not surrounded with a fibrillar plate or leuko�cytes.

In the PAS reaction, MMCs were stained in differ�ent shades of red that hardly changed after enzymatictreatment, which indicated the presence of neutralmucopolysaccharides in them. In the fish from LakeKotokel, cells in MMCs had the signs of necrobiosisand necrosis. Some of these cells had no nuclei, andothers had the signs of degenerative changes in thenuclei (karyorrhexis, karyolysis, and karyopyknosis).The number of MMCs in the spleen of the roach fromthe control water body was relatively small (on aver�age, 9.14 ± 0.62 per 1 mm2) (table). In small lightMMCs, the lymphocytic bank was absent or formed athin layer of two or three cells (Fig. 2c).

DISCUSSION

Circulatory disorders, diffuse fatty degeneration,and the presence of foci of fatty degeneration andnecrosis of hepatocytes in the liver of roach in the iceperiod in 2009 (Pronina and Pronin, 2009) indicatethat the toxin that caused the mass death of fish inLake Kotokel has a strong hepatotoxic effect. It wasalso established that the toxin has a strong hepatotoxiceffect in albino mice (Pronina et al., 2010). Presum�

(а)

(b)

(c)

(d) Fig. 1. Liver of the roach Rutilus rutilus from (a) ChivyrkuiBay and (b–d) Lake Kotokel: (a, b) melanomacrophagecenters (→), (c) erythrostasis in the sinusoids and vacuolardegeneration of hepatocytes; and (d) hemosiderin gran�ules (→) in the cytoplasm of hepatocytes. Staining: (a–c)with hematoxylin–eosin, (d) according to Pearls. Magnifica�tion: (a–c) ×200, (d) ×1000. Scale: (a–c) 50 μm, (d) 10 μm.

22



ably, such a toxin in the lake could be represented bythe microcystin of cyanobacteria (blue�green algae).Microcystis spp. and Anabaena lemmermannii, whichproduce microcystins RR, LR, and YR in the ratio49.0 : 42.5 : 8.5, were isolated from the phytoplanktonof the water body (Belykh et al., 2011). Among these,the most toxic microcystin is LR, which causes seriousdisturbances primarily in the liver (Atencio et al.,2008). However, the analysis of published data on thecases of APTM showed that many authors are skepti�cal about the role of cyanobacteria in the occurrenceof this disease (Arshanitsa et al., 1989). This was con�firmed by the experiment on feeding roaches withcyanobacteria M. aeruginosa (Kamjunke et al., 2002),which showed that microcystin had no toxic effects onfish.

The results of our study showed that the micro�structures of the liver of roaches recovered one yearafter the APTM outbreak in Lake Kotokel. It wasexperimentally established (Fournie and Courtney,2002) that an exposure to microcystin for one daycaused severe pathologic changes, including the deathof a large number of hepatocytes, in the liver of the

catfish Arius felis. Three days after the exposure to thetoxin, regenerative processes in the liver of A. felis wereobserved, and on day nine the organ recovered almostcompletely. A similar process could take place in theroaches from Lake Kotokel. However, the presence ofdeep pathological changes in the liver of one of thefishes and the presence of focal changes in themicrovasculature and parenchyma of the remainingfishes indicate the presence of a toxic component inthe water of Lake Kotokel one year after the Haff dis�ease outbreak.

The factors that caused the reduction in the areaand, respectively, the size of MMCs in the liver of theroach from Lake Kotokel remain unclear. Typically, anexposure of fish to pollutants causes an increase in thesize of MMCs, melanin accumulation, and inflamma�tory responses (Barker et al., 1994; Blazer et al., 1994;Couillard and Hodson 1996; Boorman et al., 1997;Meinelt et al., 1997; Fournie et al., 2001; Syasina,2011). However, Payne and Fancey (1989) observedsevere degenerative changes and a decrease in thenumber of MMCs in the liver of flounders Pseudopleu�ronectes americanus kept in an aquarium with high lev�

Characteristics of melanomacrophage centers (MMC) of the liver and spleen of the roach Rutilus rutilus from Lake Kotokeland Chivyrkui Bay of Lake Baikal

Parameter Kotokel Lake Chivyrkui Bay, Lake Baikal

Liver

Number of MMC, pcs./mm2

MMC area, µm2 × 10–3/mm2

Pigments, number of points:

– hemosiderin

– lipofuscin

– melanin0

Spleen

Number of MMC, pcs./mm2

MMC area, µm2 × 10–3/mm2

Pigments, number of points:

– hemosiderin0

– lipofuscin

– melanin

Mean value and error are shown above the line, and limits of variation of parameter are shown below the line. The asterisk marks the dif�ferences that were significant at p ≤ 0.05.

2 2

12.40 0.99±

8–20�� 12.06 1.22±

5–20��

7.75 0.03*±

0.09–2.80�� 11.80 0.06*±

1.09–10.68��

1.15 0.18±

0–4�� 1.25 0.14±

0–4��

3.66 0.24*±

1–6�� 2.88 0.23*±

0–7��

0.48 0.13*±

0–5��

21.20 1.16*±

13–31�� 9.14 0.62*±

4–13��

127.06 0.35*±

0.50–49.63�� 41.21 0.25*±

0.70–16.45��

11.74 1.25±

0–50��

5.26 0.57*±

0–29�� 3.22 0.6*±

0–10��

0.70 0.13*±

0–5�� 5.77 0.87*±

2–19��

108


PRONINA et al.

els of polycyclic aromatic hydrocarbons in water. Theyassumed that a high level of pollution causes adecrease in the phagocytic activity of macrophagesand the immune system of fish, leading to the degen�eration of MMCs.

The spleen of roaches from Lake Kotokel differsfrom that of fish from Lake Baikal mostly in the char�acteristics of MMCs. A significant increase in the con�tent of hemosiderin in the spleen indicates anincreased degradation of erythrocytes and the releaseof iron, which could be the consequence of damage ofthese cells by the toxin present in the water body.Hemosiderin in MMCs was detected in the fish withhemolytic anemia (Roberts, 2001) and in fish living inwater polluted with dioxin (Van der Weiden et al.,1994). According to Bucke et al. (1992), an increasedcontent of hemosiderin in the MMCs of fish spleen isthe most significant criterion for detection of waterpollution.

The increase in the content of lipofuscin in MMCsin the spleen and liver of the roach from Lake Kotokelcan probably be explained by the development ofstructural and functional abnormalities in the organsof fish. According to published data, lipofuscin canaccumulate in fish MMCs with age and in the case oftissue degeneration (Agius, 1981) and in bacterial andviral diseases and exposure to toxins (Agius and Rob�erts, 2003). However, the decrease in the content ofmelanin in the MMCs of the roaches from LakeKotokel is difficult to explain.

According to published data, MMCs concentratemacrophages after active phagocytosis, which containdead cells, pigment melanin, lipofuscin and hemosid�erin granules (Agius and Agbede, 1984; Agius, 1985),lipid droplets, and other substances with a positivestaining for the basic protein and neutral muco�polysaccharides (Herraez and Zapata, 1986). Accord�ing to many researchers (Ferguson, 1976; Ellis, 1980;Herraez and Zapata, 1986; Agius and Roberts, 2003),the main function of MMCs is the accumulation ofcellular debris and potentially toxic tissue products,

such as melanin, free radicals, and products of cata�bolic degradation in detoxification or processing ofendogenous and exogenous materials.

The number and morphology of MMCs maychange in various pathological states of the organism:starvation, inflammatory and immunological pro�cesses, viral infections, parasitic infestations, andchanging environmental conditions (Ellis et al., 1976;Wishkovsky et al., 1989; Syasina, 2011). The increasein the number and size of MMCs in the spleen of theroach from Lake Kotokel compared to the controlwater body and the presence of destructive cells andnuclei in them confirm the existing view of their sig�nificant protective role, which consists in the accumu�lation, retention, and processing of the products ofcatabolic degradation of cellular and tissue structuresthat can be hazardous for the normal functioning oforgans and organism.

In fish spleen, unlike in mammalian spleen, lym�phoid tissue is poorly developed and is located prima�rily diffusely or forms small clusters around arterioles(Ellis, 1980; Agius and Roberts, 1981; Zapata, 1982;Fänge and Nilsson, 1985; Espenes et al., 1995). Theviews of researchers about the functions of the spleenin fish are discrepant. Zapata (1982) does not considerthe spleen as an essential organ of the immune systemof fish, appealing to the fact that splenectomy has noeffect on the production of antibodies against bacteriain some fish species. At the same time, there is a stand�point that the spleen of teleost fish is the main organ oferythropoiesis and thrombopoiesis (Mikryakov andBalabanov, 1979) and the deposit of blood (Zolotov,1989; Grevati, 1991). A number of studies (Anderson,1990; Rice et al., 1996) showed that the spleen of fishis involved in the immune responses to pollutants.

The results of our study showed that lymphoid tis�sue in the spleen of the roaches from the control waterbody (Chivyrkui Bay, Lake Baikal), indeed, is poorlydeveloped, is located diffusely, and forms small shape�less clusters. The amount of lymphoid tissue in thespleen of the roaches from Lake Kotokel is much

(а) (b) (c)

Fig. 2. Spleen of the roach from (a, b) Lake Kotokel and (c) Chivyrkui Bay: (a) accumulation of hemosiderin pigment in the mel�anomacrophage center (→); (b, c) melanomacrophage center (→). Staining: (a) according to Pearls, (b, c) with hematoxylin–eosin. Magnification: ×1000; scale, 100 μm.



higher. Leukocytes form a large number of aggrega�tions and surround medium�size and large MMCswith a dense wide ring.

The increase in the number of MMCs and amountof lymphoid tissue in the spleen of the roaches fromLake Kotokel relative to the roaches from the controlwater body testifies to its active involvement in thedefense responses against exogenous toxic substances.The accumulation of small lymphocytes near MMCsindicates their direct association with the immuneresponses of this organ.

Thus, the results of the comparative study of thecharacteristic features and morphology of MMCs inthe liver and spleen of the roaches from Lake Kotokelone year after the Haff disease outbreak in Baikal ledus to the following conclusions. Since MMCs are bio�indicators of the impact of not only anthropogenicpollutants but also pathogens of natural origin on theorganism, the death and found abnormalities could becaused by cyanobacterial toxins. For assessing thedegree of intoxication of roaches in Lake Kotokel,quantitative and qualitative characteristics of MMCsin the spleen are more indicative than those in the liver.The histopathological changes observed in the liverand the response of MMTS in the spleen of roachesare indicative of the persistence of the toxin in thewater body even one year after the mass death of fish inLake Kotokel.

ACKNOWLEDGMENTS

The study in Lake Kotokel was supported by theMinistry of Natural Resources and Environment ofthe Buryat Republic.

REFERENCES

Agius, C., Preliminary studies on the ontogeny of the mel�ano�macrophages of teleost haemopoietic tissue and agerelated changes, Dev. Comp. Immunol., 1981, vol. 5,pp. 597–606.

Agius, C., The melano�macrophage centres in fish: areview, in Fish Immunology, Manning, M.J. and Tatner,M.F., Eds., London: Academic Press, 1985, pp. 85–105.

Agius, C. and Agbede, S.A., Electron microscopical studieson the genesis of lipofuscin, melanin and haemosiderin inthe haemopoietic tissues of fish, J. Fish. Biol., 1981, vol. 24,pp. 471–488.

Agius, C. and Roberts, R.J., Effects of starvation on themelano�macrophage centres of fish, J. Fish Biol., 1981,vol. 19, pp. 161–169.

Agius, C. and Roberts, R.J., Melano�macrophage centersand their role in fish pathology, J. Fish Dis., 2003, vol. 26,pp. 499 – 509.

Anderson, D.P., Immunological indicators: effects of envi�ronmental stress on immune protection and disease out�breaks, Amer. Fish. Soc. Symp., 1990, vol. 8, pp. 38–51.

Arshanitsa, N.M., Onishchenko, L.S., and Voronin, V.N.,Data on pathological and epizootic studies on the Sartlan

disease in fish, Sb. Nauch. Tr. GosNIORKh, 1989, no. 293,pp. 113–129.Atencio, L., Moreno, I., Prieto, A.I., et al., Acute effects ofmicrocystins phosphatase activities and pathologicalchanges in intraperitoneally exposed tilapia fish (Oreocho�mis sp.), Toxicol. Pathol., 2008, vol. 36, pp. 449–458.Avtandilov, G.G., Meditsinskaya morfometriya: rukovodstvo(Medical Morphometry: A Guide), Moscow: Meditsina,1990.Barker, D.E., Khan, R.A., Lee, E.M., et al., Anomalies insculpins (Myoxocephalus spp.) sampled near a pulp andpaper mill, Arch. Environ. Contam. Toxicol., 1994, vol. 26,pp. 491–496.Bazova, N.V., Matafonov, D.V., and Pronin, N.M., On thestructural changes in the communities of benthic inverte�brates of Lake Kotokel (Lake Baikal basin), Vestn. Buryat.Gossel’khozakad., 2010, no. 19, pp. 101–106.Belykh, O.I., Sorokovikova, E.G., Fedorova, G.A., et al.,Presence and genetic diversity of microcystin�producingcyanobacteria in Lake Kotokel (Russia, Lake Baikalregion), Hydrobiologia, 2011, vol. 671, pp. 241–252.Bioproduktivnost’ evtrofnykh ozer Irkana i Kotokel’ basseinaozera Baikal (Bioproductivity of Eutrophic Lakes Irkan andKotokel of the Lake Baikal Basin), Sb. Nauch. Tr. Gos�NIORKh., 1988, vol. 279.Blazer, V.S., Facey, D.E., Fournie, J.W., et al., Macrophageaggregates as indicators of environmental stress, in Modulatorsof Fish Immune Responses, Stolen, J.S. and Fletcher, T.C.V.,Eds., Fair Haven, New Jersey: SOS Publ., 1994, vol. 1,pp. 169–185.Boorman, G.A., Botts, S., Bunton, T.E., et al., Diagnosticcriteria for degenerative, inflammatory, proliferative non�neoplastic and neoplastic liver lesions in medaka (Oryziaslatipes): consensus of a national toxicology program pathol�ogy working group, Toxicol. Pathol., 1997, vol. 25, no. 2,pp. 202–210.Bucke, D., Vethaal, A.D., and Lang, T., Quantitative assess�ment of melanomacrophage centres (MMCs) in dabLimanda limanda along a pollution transect in the Germanbight, Mar. Ecol. Progr. Ser., 1992, vol. 91, pp. 193–196.Couillard, C.M. and Hodson, P.V., Pigmented macrophageaggregates: a toxic response in fish exposed to bleached�kraft mill effluent?, Environ. Toxicol. Chem., 1996, vol. 15,pp. 1844–1854.Ellis, A.E., Munroe, A.L.S., and Roberts, R.J., A study ofthe phagocytic system and the fate of intraperitoneallyinjected particulate material in the plaice (Pleuronectes pla�tessa L.), J. Fish. Biol., 1976, vol. 8, pp. 67–78.Ellis, A.E., Antigen�trapping in the spleen and kidney of theplaice, Pleuronectes platessa (L.), J. Fish Dis., 1980, vol. 3,pp. 413–426.Espenes, A., Press, C.McL., Dannevig, B.H., and Lands�verk, T., Investigation of the structural and functional fea�tures of splenic ellipsoids in rainbow trout (Oncorhynchusmykiss), Cell Tiss. Res., 1995, vol. 279, pp. 469–474.Fange, R. and Nilsson, S., The fish spleen: structure andfunction, Experientia, 1985, vol. 41, pp. 152–158.Ferguson, H.W., The relationship between ellipsoids andmelano�macrophage centres in the spleen of turbot (Scoph�thalmus maximus), J. Comp. Pathol., 1976, vol. 86, pp. 377–380.

110


PRONINA et al.

Fish pathology, Roberts, R.J., Ed., Philadelphia, PA: W.B.Saunders, 2001.Fournie, J.W. and Courtney, L.A., Histopathological evi�dence of regeneration following hepatotoxic effects of thecyanotoxin MC�LR in the hardhead catfish and gulf killi�fish, J. Aquat. Anim. Health, 2002, vol. 14, pp. 273–280.Fournie, J.W., Summers, J.K., Courtney, L.A., et al., Util�ity of splenic macrophage aggregates as an indicator of fishexposure to degraded environments, J. Aquat. Anim. Health,2001, vol. 13, pp. 105–116.Grevati, A.D., Electron�microscopic study of cells andblood�forming organs of the mirror carp, Extended Abstractof Cand. Sci. (Biol.) Dissertation, Moscow: Mosk. Gos.Univ., 1991.Haaparanta, E., Valtonen, T., Hoffmann, R., and Holmes, J.,Do macrophage centres in fresh water fishes reflect the dif�ferences in water quality?, Aquat. Toxicol., 1996, vol. 34,pp. 253–272.Herraez, M.P. and Zapata, A.G., Structure and function ofthe melano�macrophage centres of the goldfish Carassiusauritus, Vet. Immunol. Immunopathol., 1986, vol. 12,pp. 117–126.Kachina, E.A. and Komarova, E.V., Analysis of the clinicalpicture, diagnosis, and treatment of Haff disease in theRepublic of Buryatia, in Mater. student. nauch.�prakt. konf(Proc. student Sci.�Pract. Conf.), Krasnoyarsk: Verso,2009, pp. 314–315.Kamjunke, N., Schmidt, K., Pelugmacher, S., and Menher, T.,Consumption of cyanobacteria by roach (Rutilus rutilus):useful or harmful to the fish?, Freshwat. Biol., 2002, vol. 47,pp. 243–250.Khan, R.A., Barker, D.E., Hooper, R., et al., Histopathol�ogy in winter flounder (pleuronectes americanus) livingadjacent to a pulp and paper mill, Arch. Environ. Contam.Toxicol., 1994, vol. 26, pp. 95–102.Mackmull, G. and Michels, N.A., Absorption of colloidalcarbon from peritoneal cavity in the teleost Tautogolabrusadspersus, Amer. J. Anatom., 1932, vol. 51, pp. 3–47.Meinelt, T., Krueger, R., Pietrock, M., et al., Mercury pol�lution and macrophage centers in pike (Esox lucius) tissues,Environ. Sci. Pollut. Res., 1997, vol. 4, pp. 32–36.Merkulov, G.A., Kurs patogistologicheskoi tekhniki(A Course of Pathohistological Technique), Leningrad:Meditsina, 1969.Mikryakov, V.R. and Balabanova, L.V., Cellular basis of fishimmunity, in Fiziologiya i parazitologiya presnovodnykh zhi�votnykh (Physiology and Parasitology of Freshwater Ani�mals), Leningrad: Nauka, 1979, pp. 57–64.Payne, JF. and Fancey, L.F., Effect of polycyclic aromatichydrocarbons on immune responses in fish: changes in mel�ano�macrophage centers in flounder (Pseudopleuronectesamericanus) exposed to hydrocarbon�contaminated sedi�ments, Mar. Environ. Res., 1989, vol. 28, pp. 431–435.Pintucci, G., Manzionna, M., Maida, I., et al., Morpho�functional characterization of cultured pigment cells fromRana esculenta L. liver, In Vitro Cell Devel. Biol., 1990,vol. 26, pp. 659–664.Pronin, N.M., Chivirkui Bay in Lake Baikal as super�unique open ecosystem for complex interdisciplinaryresearch, Abstr. IIIth Vereschagin Baikal Conf., Irkutsk,2000, p. 188.

Pronina, S.V. and Pronin, N.M., Pathological changes inthe liver of roach Rutilus rutilus from Lake Kotokel (Baikal)during the Haff disease outbreak, Sb. Nauch. Tr. Gos�NIORKh, 2009, no. 338, pp. 170–172.Pronin, N.M., Matveev, A.N., Samusenok, V.P., et al., Rybyozera Baikal i ego basseina (Fishes of Lake Baikal and ItsBasin), Ulan�Ude: Buryat. NTs SO RAN, 2007.Pronin, N.M., Burdukovskaya, T.G., Sondueva, L.D.,et al., Parasite fauna and features of the epizootic situationin the roach population in Lake Kotokel, Sb. Nauch. Tr.GosNIORKh, 2009, no. 338, pp. 171–173.Pronin, N.M., Burdukovskaya, T.G., Batueva, M.D., et al.,Parasite fauna of the perch in Lake Kotokel (Republic ofBuryatia: Baikal region) during the Haff disease outbreak,Vestn. BuryatGU, Ser. Biol. Georg., 2010a, no. 4, pp. 174–179.Pronina, S.V., Pronin, N.M., Shantanova, L.N., and Mon�dodoev, A.G., Pathomorphological changes in the tissues ofwhite mice fed with fish from Lake Kotokel during the Haffdisease outbreak, Byul. Vost.�Sib. NTs SO RAMN, 2010b,no. 3 (73), pp. 256–260.Rice, C.D., Kergosien, D.H., and Adams, M., Innateimmune function as a bioindicator of pollution stress infish, Ecotoxicol. Environ. Safety, 1996, vol. 33, pp. 186–192.Roberts, R.J., Melanin�containing cells of the teleost fishand their relation to disease, in The Pathology of Fishes,Ribelin, W.E. and Migaki, G., Eds., Madison: Univ. Wis�consin Press, 1975, pp. 399–428.Scalia, M., Geremia, E., Corsaro, C., et al., The extracuta�neous pigmentary system; evidence for melanosynthesis inAmphibia and Reptilia liver, Comp. Biochem. Physiol., 1988,vol. 85, pp. 715–717.Syasina, I.G., Characteristics of histopathological changesin organs of banded catfish Pelteobagrus fulvidraco(Bagridae) from the Amur River basin, J. Ichthyol., 2011,vol. 51, no. 3, pp. 263–276.Van der Weiden, M.E.J., Bleumink, R., Seinen, W., and Vander Berg, M., Concurrence of P450 1A induction and toxiceffects in the mirror carp (Cyprinus carpio), after adminis�tration of a low dose of 2,3,7,8�tetrachlorodibenzo�p�dioxin, Aquat. Toxicol., 1994, vol. 29, pp. 147–162.Wishkovsky, A., Mathews, E.S., and Weeks, B.A., Effect oftributyltin on the chemiluminescent response of phagocytesfrom three species of estuarine fish, Arch. Environ. Contam.Toxicol., 1989, vol. 18, pp. 826–831.Wolke, R.E., Murchelano, R.A., Dickstein, C., andGeorge, C.J., Preliminary evaluation of the use of mac�rophage aggregates (MA) as fish health monitors, Bull.Environ. Contam. Toxicol., 1985, vol. 35, pp. 222–227.Wolke, R.E., Piscine macrophage aggregates: a review, Ann.Rev. Fish. Dis., 1992, vol. 2, pp. 91–108.Zapata, A., 1982. lymphoid organs of teleost fish. iii. spleniclymphoid tissue of rutilus rutilus and gobio gobio, Devel.Comp. Immunol, vol. 6, pp. 87–94.Zolotova, T.E., 1989. Eksperimental’noe issledovaniekrovetvoreniya u ryb: Avtoref. dis. … kand. biol. nauk(TRANSLATION), Moscow: MGU.Zuasti, A., Jimenez�Cervanes, C., Garcia�Boron, J.C., andFerrer, C., 1998. the melanogenic system of xenopus laevis,Arch. Histol. Cytol, vol. 61, pp. 305–316.

Translated by M. Batrukova

2 2

22

22

SPELL: 1. dystrophication, 2. Rutilus, 3. Percae

Characteristics of Melanomacrophage Centers in the Liver and Spleen of the Roach Rutilus rutilus (Cypriniformes: Cyprinidae) in Lake Kotokel during the Haff Disease Outbreak

Documents