Umbra Krameri

4. В.Зенкин, Е.Рязанцева, О.Лосев „Полиморфизм мышечных эстераз и анализ популяционной структуры обыкновенной и капской ставрид шельфа западной Африки” (Биохимическая и популяционная генетика рыб), Ленинград,1979. c.94 5. В.Кирпичников “Генетика и селекция рыб”, Ленинград, 1987. pag.519 6. Л.Смирнов “Сравнительная оценка белковых спектров печени и мускулатуры рыб, птиц и млекопитающих, получаемых методом диск-электрофореза в полиакриламидном геле” (Сравнительная биохимия рыб и их гельминтов) Петрoзаводск, 1977. Pag.85 7. Л.Шарт, Ю.Илясов „О типах трансферpинов и эстераз у производителей карпа селекционируемых нa устойчивость к краснухе”, (Биохимическая и популяционная генетика рыб), Ленинград, 1979. Pag.147

TWO NEW CNIDOSPOREAN SPECIES (CNIDOSPORA: SPHAERO- SPORIDAE, MYXOBOLIDAE), PARASITES OF THE EUROPEAN MUDMIN- NOW (UMBRA KRAMERI) FROM LOWER DNIESTER RIVER Alexander Moshu, Ilya Trombitsky* Institute of Zoology, Academy of the Sciences of Moldova; *Fisheries Research Station Introduction The European mudminnow – Umbra krameri Walbaum, 1792 (Esociformes: Umbridae) from the zoogeographical point of view is as a relict endemic of the Danube and Lower Dniester River basins and of the Black Sea region of Ponto-Aralo-Caspian province of Holarctic [1]. At present this freshwater fish is highly rare and figures in Lists of the Red Books of the World, Europe, Romania, Republic of Moldova and Ukraine as an especially threatened species [13]. The analysis of the collected material showed, that parasitic fauna of U.krameri is rich and diverse (about 45 species of different taxonomical groups) and is included on the whole with the common and widely-distributed species of the Dniester River and other waterbodies of the region. Among them only nine species seem to be more or less specific for this fish and pike (Esox lucius L., 1758) [28]. Conforming opinion of parasitologists [24, 26, 27, 31], parasitic fauna of the relict endemic fish are mostly not specific. However, among European mudminnow parasites we discovered, several species prove to be unknown to science [23, 28]. The description of two of them is presented below. Materials and Methods

The host specimens from the lower flow of the Dniester basin (Cuciurgan reservoir - South-Eastern part of the Republic of Moldova; Turunciuc branch and Putrino Lake - South-Western part of Ukraine) were collected between February and March of 2000 [21]. The data was obtained through a complete parasitological investigation of 30 specimens of U.krameri resulting from different ages - 1-5 years old, sizes - 7,2 (5,5-9,5) cm length, and genders - 19 males and 11 females. In addition, 34 fish fixed in 4% formaldehyde (5.0-7.0 cm long) from the middle Danube basin (Cernica Lake - near Bucharest, Romania) in May 2003 were examined. The material was routinely processed according to standard ichthyoparasitologi-

78 cal procedures [8, 11, 22, 29, 30]. Description, measurements (in micrometers) and drawings of parasites were taken under phase-contrast optics from fresh or preserved smears in glycerine-jelly and are based upon no less than 50 spores. Infected tissues were fixed in buffered 4% neutral formaldehyde. The identified parasites were compared with those from slides of Sphaerospora and Myxobolus species from the Pro- tozoological Collection at the Institute of Zoology of the Academy of the Sciences of Moldova (Chisinau) and at the Zoological Institute of the Russian Academy of Sci- ences (Saint-Petersburg). Results and Discussions

Description of the species Phylum Cnidospora Doflein, 1901, Class Myxosporea Buetschli, 1881, Order Bivalvulida Schulman, 1959, Suborder Varrisporina Lom et Noble, 1984, Family Sphaerosporidae Davis, 1917, Genus Sphaerospora Thélohan, 1892 Species Sphaerospora umbrae, sp. n. (Fig. 1)

Fig. 1. Line-drawing of the Sphaerospora umbrae sp. n. spores from kidney of the European mudminnow, Umbra krameri (bar = 5 µ)

Host: European mudminnow - Umbra krameri Walbaum, 1792. Age: 2-5. Locality: lower flow of the Dniester basin (Cuciurgan reservoir - South-Eastern part of the Republic of Moldova; Turunciuc branch and Putrino lake - South-Western part of Ukraine), February-March 2000; middle Danube basin (Cernica lake - South- Eastern part of Romania), May 2003. Site of infestation: inside the Bowman’s capsule, in the lumens and epithelia of renal tubules, in ureteres and urinary bladder. Early presporogonic developmental stages of parasite, more often - in the blood of kidney (its head portion) and gills, rarely – in the blood of choroidal and ocular rete mirabile, spleen and peripheral vessels. Prevalence and intensity of invasion: Plasmodia and pseudoplasmodia (= pansporoblasts) - in 22 out of 30 (73,3%) examined specimens, spores - in 9 specimens (30%) only. Intensity of invasion of fish was very low (up to 14 spores in a field of view of a microscope, 7x40). Blood presporogonic stages of parasite development in all examined fish were found (kidney - 27/30, gills -23/30, rete mirabile - 4/30, spleen - 3/30, peripheral stream - 1/30).

79 Description: Vegetative stages are large round-oval or oval-elongate shaped mother plasmodia, 25.0-30.0 x 35.0-50.0 µ in size, with large-granulated endoplasm and slight distinct thin ectoplasm, which forms at the edge several short lobopodia. Within each plasmodium 4-8 rounded (8.75-17.5 µ in size) to oval (6.87-8.5 x 10.0- 12.5 µ in size) disporous pseudoplasmodia are formed. They also contain the refractive inclusions and droplets, black and yellow coloured granules. The plasmodia and pseudoplasmodia in lumen of the renal tubules can be seen solitary or in conglomerations, arranged free or, rarely - attached by rootlike shoots of ectoplasm to the epithelia. Para- site development was asynchronous. Mature and young plasmodia, as well as pseudoplasmodia, occurred in the same tubular portion. Moreover, the plasmodia contained spores in varying states of maturity at the same time. In vital blood samples from the kidney and the gills, but rarely in other organs, the immobile rounded multinucleate bodies were detected (6.2-10.0 µ in diameter) with granulated endoplasm, sometimes having fringed ectoplasm. These bodies, revealed often by other authors for Sphaero- spora invasion of different fish species, are considered as early proliferative stages of parasitic development [3, 4, 9, 11, 12, 15, 16]. In sutural view the mature spores are almost subspherical, with a somewhat prominent anterior end (on apex of each valve a short thickening are observed), where distinct suture bulge forms a small short knob on each valve near suture and in posterior rounded end bulge form a well visible keel. Valves are thick-walled, its surface is striated (3-4 well-distinguished longitudinal fine rows on the posterior end of each valve, which oriented, and meridionally, especially well distinguished on the posterior surface of spores like as smalls ridges), which comes off in the middle of spore light (fine ridges running anteriorly). The sutural line is slightly prominent and suture margin not run out. Spores measure 6.25 (4.75-6.87) µ long, 6.2 (6.0-6.25) µ wide and 6.25 (3.75-6.87) µ thick. Each spore contains two slight distinct, oval-pyriform shapes and equally sized extrusive capsules, 1.2-2.5 µ in size, which are positioned at some distance from anterior apex of spore and opens on each side of suture margin. The extrusive filaments with 3-3.5 coils, are arranged almost parallel or 45о to the transverse axis of capsule. Extended filament is comparatively thick and measures up to 8.5-8.8 µ in length. The transversally elongate and binuclear sporoplasm occupies all remained volume of the spore. The collected material shows the presence of specific polymorphism at this species, which is expressed by a high variability of the morphometrical data of the struc- tures (vegetative forms and spores) and it depends a great deal on the maturity of the parasite and the intensity of the kidney’s invasion. The juvenile spores, unlike mature ones, are bigger, round or broadly shaped (in valvular view), with a clearly expressed and projected suture margin, with non-smooth and thicker valves, on posterior surface of which the slightly expressed knobbly was observed. Occasionally the lateral-proximal surface of each shell valve of the maturing spores, opposite the sutural plane, appeared to have a small lateral ear-like process and on the proximal surface – surrounded by mucous envelope. On the whole, valves architectonic are more pronounced in immature spores. Differential diagnosis and remarks: Previously no Sphaerospora species had been reported from U.krameri. Based upon the morphometrical characteristics the species in question finds out some looking alike to S.minuta Konovalov, 1967, first de-

80 scribed from ureteres and urinary bladder of the common pike fingerlings (Penjina river - border between the Chukot and the Kamchatka, North-Eastern part of Russia) [30] and was repeatedly found by us throughout pikes in Moldova’s waterbodies (Prut and Dniester basins, 1988-1995). Moreover, S.umbrae sp. n. clearly differs from S.minuta by fish-host, the spores less oval shape, the variability of the valves ornamental pattern (not pronounced suture and their margin, less thick valves, presence of striation on the rounded posterior pole of spores), by smaller length of spores, and by the smaller extrusive capsules and their arrangement inside the spores (Fig. 2 and Tab. l).

Fig. 2. Sphaerospora minuta Konovalov, 1967 from the kidney of common pike (Esox lucius L.) from Moldova’s waterbodies (bar = 5 µ) The comparison with sphaerosporean species of Salmoniformes fish -S. rota Za- ika, 1961, S.krogiusi (Konovalov et Schulman in: Schulman, 1966), S. truttae Fischer- Scherl, El-Matbouli et Hoffmann, 1986 and S. coregoni El-Matbouli, Hoffman et Kern, 1995 [5, 6, 30, 32] has shown that discovered in U. krameri parasite is distinguished in the structure and sizes of the developmental stages, sporal and capsular shapes and di- mensions, pattern of valvular ornamentation. Besides different fish-hosts, other known species are morphologically different from it in other respects. From all compared Sphaerospora spp. described parasite differs in having the blood stages of development, which can be explained by insufficient studying of fish in this aspect before. Table. Size characteristics of S. umbrae and S. minuta spores (µ)

S.minuta S.minuta S.umbrae, sp. n. (Moldova’s waterbodies) (Penjina river) Spores 6.25 (4.75-6.87) long 6.0-7.3 long 6.2-7.3 long 6.2 (6.0-6.25) wide 5.7-6.3 wide 5.8-6.8 wide 6.25 (3.75-6.87) thick 5.7-6.8 thick 5.0 thick Extrusive 2.9-3.6 long 2.9-3.3 long capsules 2.25-2.5 in size 2.4-2.7 wide 2.6-2.8 wide

It should be noted, that S.minuta discovered in pikes of Moldova’s waters also has certain distinctions from its original description and as well as specimens of parasite in type-slides contained in the Protozoological collection of the Zoological Institute at the Russian Academy of Sciences (slide Nr.319, Sph.minuta, Myxobolus musculi, det. Konovalov, Esox lucius, age 0+, kidney, Penjina river, 18.08.1966. coll. Konovalov) (Tabl.). We are prone to surmise that parasites from fish of Penjina basin, apparently, were described by immature spores. We do not exclude the possibility of existence of the separate or close populations of the same species for each of the hosts (pike and

81 mudminnow). In this connection, it is necessary to compare the parasite of pike from different regions and gives more full description of this species. Pathological effects: In the kidney of some heavily infested fish specimens besides some local dystrophic changes on histological level (the signs of hyperplasia, vacuolisation and hyalinisation of the tubular epithelium, tissue with a huge number of so called “rodlet cells” [14, 20] and in enlarged local distension lumen of tubules agglomerations of the crystals mass – nephrocalcinosis [10, 11]), no other macroscopic lesions were showed. Type material: Syntypes in type slides of described parasite (with acquisition number 38-48), as well as fixed infected tissue have been deposited in the collection of the Laboratory of Ichthyology and Aquaculture, Institute of Zoology of the Academy of Sciences of Moldova. Suborder Platysporina Kudo, 1984, Family Myxobolidae Thélohan, 1892, Genus Myxobolus Thélohan, 1892 Species Myxobolus variatus, sp. n. (Fig. 3) Host: European mudminnow - Umbra krameri Walbaum, 1792. Age: 2-5. Locality: lower flow of the Dniester River basin (Cuciurgan reservoir - South- Eastern part of Republic of Moldova; Turunciuc branch and Putrino lake - South-West- ern part of Ukraine), February-March 2000; middle flow of the Danube basin (Cernica lake - South-Eastern part of Romania), May 2003. Site of infestation: muscle (in sarcoplasm of muscle fibres and along them); kidney parenchyma (in the melanomacrophages and limfohaemopoetic centres of the trunk kidney); dispersed spores - in the gills, gut, gall bladder and occasionally - in liver-pancreas.

Fig. 3. Line-drawing of the Myxobolus variatus sp. n. frequently founded spores in muscle and kidney of European mudminnow, Umbra krameri (bar = µ) Prevalence and intensivity of invasion: in 6 out of 30 (20%) examined specimens from Dniester River basin and in 29 out of 34 (73,3%) specimens from Danube River basin were found to be infested. The level of intensity of invasion of fish varied widely but was usually low. Description: Vegetative stages of this parasite present the oval-elongate or sometimes irregularly shaped multinucleate mother-trophozoites (=plasmodia), up to 180.0 µ in size. Its protoplasm is slightly differentiated, having fine-granulated endoplasm, packed with numerous small refractive droplets/inclusions, sometimes (in kidney) black or yellow coloured granules and surrounded with thin layered and transparent

82 ectoplasm, which does not form pseudopodia-like processes. Within each plasmodium usually 4-8 or more rounded-oval (measured 12.5-23.2 µ in size) disporous pansporoblasts were formed. They can be seen in groups (usually with 2-4 specimens and more) in muscles and kidney or solitary - in other sites. The typical and mature spores in valvular view were of different shapes: the ma- jority are almost oval-ovoid shaped - commonly with slightly wider-rounded posterior pole than anterior one and vice versa. Usually in valvular view the spores are slightly asymmetrical by convex sides. In sutural view, spores are not varying. Valves are thick- walled (especially on the poles of the spore), with smooth surface, do not form by sutural thick intercapsular process or edge radial markings on the distal pole. The line suture between valves is straight and slightly prominent. In sutural view, suture mar- gins of valves are relatively wide, well expressed and protrude on both narrowing poles of spores like a small keel (more relieved at anterior pole and less at posterior one). The length of spores is 9.6 (8.8-11.2) µ, its width 6.4 (5.6-8.0) µ and thickness 6.0 (5.6-6.7) µ. Each spore contains two pyriform shaped and equal in size extrusive capsules. They are apically positioned in spore and occupy up to half of length of them. Their anterior slightly tapering ends converge and proximal ones are distant. Each capsule through a foramen in valve opens side by side (on each side near thickenings of suture) at the anterior pole of the spore. The extrusive capsules length is 4.8 (3.5-4.8) µ, its thickness 2.4 (1.9-2.9) µ. The extrusive filaments are coiled with 6 (5-7) turns (it is more evident in juvenile spores), oriented roughly 45o (rarely perpendicular) to the longitudinal axis of the capsule. The extended filaments are thin and measure 24.0 (19.2-27.2) µ long. In- side of the spores the fine-grained binuclear sporoplasm, with one round to oval shaped well-distinct iodinophillous vacuole and two small rests of capsulogenic/valvologenic nuclei lying near the posterior part of the capsules, were observed. The collected material shows the high variability of the morphometrical data of the spores, which depends a great deal on their maturity. The maturing spores are differentiated in size, rounded or broadly shaped in valvular plane, with thicker valves, clearly expressed suture margin, occasionally with non-smooth shell surface on the posterior end (surrounded by thin transparent extern mucous envelope around the posterior end), more pyriform capsules and more evidently turned extrusive filaments. The bigger iodinophillous vacuole and two small rests of the capsulogenic/valvulogenic nuclei were frequently visible inside the juvenile spores. It was signalled also the different aberrant spores: elliptic, elongated-oval, pyriform or wide pyriform shaped, with slight asymmetric narrowed anterior pole, dilated posterior pole, displacement of the capsules laterally, one or three capsules and with one small caudal processes on the posterior shell surface of each valve, etc. Furthermore, comparison of the spores from fish caught in different waterbodies has shown only some little variations in thickness of valves and sporal sizes (e.g. the spores from Dniester have appeared a little bit more large and with thick-walled of those from Danube). Differential diagnosis and remarks: For U.krameri the data about finding of Myxobolus species in the literature are absent. Based upon the morphometrical characteristics of the discovered parasite finds some similarity to M.dechtiari Cone et An- derson, 1977 [2], described in gills of Lepomis gibbosus L. from Canada. However the U.krameri parasite is distinguished in the little other shape of spores, absence of sutural

83 markings, shape of capsules, absence of intercapsular appendix, other orientation of filament coils, by little turns of filament, smaller sizes of spores and length of capsules and by shorter filament. The species in question has also certain similarity with M. cf. karelicus Petruschewsky, 1940 (gills of L.gibbosus from Europe) [7], but descriptions of M.karelicus [25, 32] have shown, that spores, nevertheless, have a small intercapsular appendix. It is evident that M.karelicus spores are narrower at anterior pole in comparison with ones of M.variatus sp. n. Furthermore, according to J. Lom (1969), extrusive filament forms in capsules 8-spiralled coils. Taking in consideration the dis- covery of these parasites (M.dechtiari and M.karelicus) in filogenetically relatives fish (order Perciformes), we do not exclude the possibility that they may be the same species. From M.subepithelialis Weiser, 1949 (gills, kidney, intestinal wall of Gobio obtu- sirostris, Alburnus alburnus and Perca fluviatilis) from Dniester basin M.variatus sp. n. clearly differs by structure of vegetative stages, spores shape, thicker valves, absence of intercapsular appendix, little length and width of spores, thicker spores and smaller capsules. From all compared Myxobolus spp. described parasite differs by having in valvular view the spores asymmetrical in convex sides. Besides different fish-hosts, other known till now Myxobolus species are morphologically different from it by other features. Thus, the recorded parasite could not be identified with any of the known species of the genus and the above-mentioned differences justify description of M.variatus as a new species. Pathological effects: No macroscopic or microscopic changes due to M.variatus sp. n. were observed in infested fishes. Type material: Syntypes in type slides of described parasite (with acquisition number 49-55), as well as the fixed infected tissues have been deposited in the collection of the Laboratory of Ichthyology and Aquaculture, Institute of Zoology of the Academy of Sciences of Moldova. Acknowledgments. The authors are grateful to ichthyologist Oleg Strugulea (Institute of Zoology of the Academy of Sciences of Moldova) for supplying the fish material from Dniester basin and to Dr. Grigore Davideanu (Natural History Museum, “Al. I. Cuza” University, Iasi, Romania) for put at authors disposal the fish specimens from Danube basin. Our thanks are due to Dr. Pavel Gerasev for permission to compare the identified material with slides of Sphaerospora species from Collection of the Zoological Institute of the Russian Academy of Sciences, Prof. Dr. Oleg Pugachev (Zoological Institute of the Russian Academy of Sciences) and Prof. Dr. Ion Toderash (Institute of Zoology of the Academy of Sciences of Moldova) for critically reviewing the manuscript. References

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Articolul este prezentat de academicianul I.Toderaş

НОВЫЕ ВИДЫ РАСТЕНИЕОБИТАЮЩИХ КЛЕЩЕЙ ДЛЯ ФАУНЫ МОЛДОВЫ Л.М. Куликова Институт зоологии Академии наук Молдовы Введение Данное сообщение является частью комплексных исследований акарокомплексов биоценозов Молдовы. Видовой состав и экологические особенности некоторых групп в акарокомплексах конкретных биоценозов оставались недостаточно изученными. Избирательность клещей к питанию на определенных видах растений связана с биохимическим составом клеточного сока последних. Проведенные сборы позволили пополнить данные по видовому разнообразию и трофическим связям растениеобитающих клещей. Существенные различия в диагностических признаках (сочетание и характер изменчивости) отдельных представителей растениеобитающих клещей послужили основанием для включения их как новые виды в фауну Молдовы [19]. Материал и методика Фаунистическая инвентаризация растениеобитающих клещей на территории Республики Молдова проводилась способом полевых маршрутных обследований. В лесных массивах на растениях естественного произрастания и садах на плодовых породах, в зависимости от подвижности клещей, пробы отбирались по методике: побеги – 10 см, листья - 100 листьев (с 10 деревьев по 10 листьев). Проведена ревизия коллекции растениеобитающих клещей Института Зоологии АН Молдовы. Результаты исследований Растениеобитающие клещи составляют элемент класса Arachnida и играют существенную роль в биоценозах. По своему систематическому положению ранее выявленные растениеобитающие клещи относятся к 2 отрядам, 16 семействам, 218 видам [7]. Из фитофагов самыми многоядными являются клещи семейств Acari- dae и Tetranychidae. Список повреждаемых ими растений состоит из нескольких десятков названий. Хищные клещи представлены семействами Cunaxidae и Phyto- seiidae, обладающие высокой поисковой активностью, прожорливостью, широким кругом растений-хозяев и достаточно высокой длительностью жизни [6, 7, 8]. В результате исследований были обнаружены 16 новых видов растениеобитающих клещей для фауны Молдовы.