Hemoglobin patterns in different populations of Synbranchus marmoratus Bloch, 1795 (Pisces, Synbranchidae)

Total hemolysates of Synbranchus marmoratus Bloch, 1795 captured at four different sites in the State of S?o Paulo, Brazil, showed two different hemoglobin phenotypes when submitted to agar-starch gel electrophoresis on glass slides in basic buffer. Phenotype I was characterized by 3 hemoglobin bands. When the total hemolysate was submitted to cellulose acetate electrophoresis in basic buffer containing 6 M urea and beta-mercaptoethanol, Phenotype I showed four globins of the alpha 1, alpha 2, beta and gamma types, with 11.9 +/- 1.9 g% total hemoglobin, 45.3 +/- 3.6% globular volume, and 26.8 +/- 4.4% mean corpuscular hemoglobin concentration (MCHC). Phenotype II showed three groups of hemoglobins, with a total of up to 12 hemoglobin bands. When the total hemolysate was submitted to cellulose acetate electrophoresis in basic buffer containing 6 M urea and beta-mercaptoethanol, phenotype II showed five types of globins, denoted types alpha 1, alpha 2, gamma 1, gamma 2 and beta, having electrophoretic positions different from those of Phenotype I globins, with 18.1 +/- 3.3% total hemoglobin, 47.9 +/- 6.4% globular volume, and 37.8 +/- 4.4% MCHC. The distribution of the specimens having the two hemoglobin phenotypes is associated with the different geomorphological provinces of the State of S?o Paulo, suggesting the existence of at least two populational groups of Synbranchus marmoratus.     

Characterization of the hemoglobins and globins of Synbranchus marmoratus Bloch, 1795 (Pisces, Synbranchidae)

Total hemolysates of Synbranchus marmoratus Bloch, 1795, captured in Vitoriana, district of Botucatu, State of S?o Paulo, Brazil, were submitted to agar-starch gel electrophoresis on glass slides using 42 mM-Tris 1.7 mM EDTA-6.1 mM borate buffer, pH 8.8, for the gel and 10 mM borate-1.7 mM NaOH buffer, pH 8.6, for the cuvette. Three distinct hemoglobin bands were detected, with Hb I being of the cathodic type. Cellulose acetate electrophoresis in 800 mM Tris-2.1 mM EDTA buffer, pH 8.9, containing 6 M urea and 2.25 mM beta-mercaptoethanol indicated the presence of four globin chains denoted alpha 1, alpha 2, beta and gamma. It is suggested that the probable tetrameric constitution of the hemoglobin of Synbranchus marmoratus Bloch, 1795 is Hb I (alpha (2)2 gamma 2), Hb II (alpha (1)2 gamma 2) and Hb III (alpha (1)2 beta 2).     

Ultrastructure of the testis in Synbranchus marmoratus (Teleostei,Synbranchidae): the germinal compartment

Synbranchus marmoratus, is a protogynic diandric species in which two types of males, primary and secondary, are found. In both types, the germinal compartment in the testes is of the unrestricted lobular type, but in secondary (sex reversed females) males the lobules develop within the former ovarian lamellae. In the present study, the germinal compartment was examined in both types of males using light microscopy as well as scanning and transmission electron microscopy. Germinal compartment is limited by a basement membrane and contains Sertoli and germ cells. During maturation, processes of Sertoli cells form the borders of spermatocysts containing isogenic germ cells. Characteristically, type A and type B spermatogonia have a single nucleolus and grouped mitochondria associated with dense bodies or nuage. Type B spermatogonia, spermatocytes and spermatids are joined by cytoplasmatic bridges and are confined within spermatocysts. Secondary spermatocytes are difficult to find, indicating that this stage is of short duration. Biflagellated spermatozoa have a rounded head, no acrosome, and possess a midpiece consisting of two basal bodies, each of which produces a flagellum with a typical 9+2 microtubular composition. No associations occur between sperm and Sertoli cells. There were no differences between spermatogenesis in primary and secondary males in this protogynic, diandric fish.     

Testicular interstitial cells, and steroidogenic detection in the protogynous fish, Synbranchus marmoratus (Teleostei, Synbranchidae)

The swamp eel, Synbranchus marmoratus, is a freshwater protogynic diandric species. Primary males develop directly as males while secondary males arise from the sex reversal of females. Fishes from Argentine and Brazil inland waters were collected, examined and compared for this study. In order to characterize the interstitial testicular compartment, light and electron microscopy techniques and an enzyme histochemical examination for steroidogenic cells detection were used. The interstitial compartment of S. marmoratus is composed of Leydig and myoid cells, collagen fibers, blood cells, macrophages,and amyelinic nerves. At the ultrastructural level, no differences were observed in the interstitial tissue, either between specimens from the different sampling sites or between primary and secondary males. Leydig cells are present in all testes examined throughout the year. A cytoplasmatic reaction of 3beta-HSD was detected only in Leydig cells during sex reversal and in both type of males, mainly during the regressed and early maturation classes (autumn and winter). Leydig cells possess the typical fine structural characteristics associated with steroidogenesis. Furthermore, in both type of males, during sex reversal and after the spawning period, the number of granulocytes and macrophages present in the testes increased, suggesting that they could be involved in phagocytosis and resorption of damaged cells.     

Ovarian follicle ultrastructure in the teleost Synbranchus marmoratus (Bloch, 1795), with special reference to the vitelline envelope development

Synbranchus marmoratus is a protogynous diandric teleost fish widely distributed throughout South America. The aim of this work was to study the ultrastructure of the vitelline envelope and the relationship among oocyte and their follicular cells during oogenesis. During perinucleolar stage, the oocyte and the follicular cells form microvillar processes that project into the perivitelline space. The oocyte secretes a dense and amorphous material, which appears as the first evidence of the vitelline envelope (VE) development. The VE passes from a double to a multilayered structure during oocyte growth. In mature oocytes, the VE reach a mean thickness of 11 microm, having up to 30 layers. Oocyte microvilli are thinner than the follicular ones and were seen in contact with the follicular plasmalema, however we could not find any contact between the follicular microvilli and the oolemma. Before ovulation, microvillar processes retract and the pore canals seem to collapse. An outer electron dense layer occludes the superficial pore and forms a continuous layer. No jelly or adhesive coatings were seen at least in ovulated eggs sampled from ovarian lumen. Follicular cell and oocyte cytological characteristics do not differ from those described in other teleosts species.     

Metabolic adjustments during semi-aestivation of the marble swamp eel (Synbranchus marmoratus, Bloch 1795)--a facultative air breathing fish.

Metabolic changes, principally in intermediary metabolism and nitrogen excretion, were investigated in the marble swamp eel (Synbranchus marmoratus) after 15 and 45 days of artificially induced semi-aestivation. Glucose, glycogen, lactate, pyruvate, free amino acids, triglycerides, ammonia, urea, and urate contents were determined in liver, kidney, white muscle, heart, brain, and plasma. Lactate dehydrogenase, glutamate dehydrogenase, malate dehydrogenase, aspartate amino transferase, alanine amino transferase, glutamine synthase, ornithine carbamoyl transferase, and arginase enzymes were assayed. The teleost S. marmoratus maintained initial energetic demands by lipid oxidation. The course of normal oxidative processes was observed through tissue enzyme profiles. After the lipid stores were exhausted, the fish consumed body proteins. Constant values of hematocrit during induced semi-aestivation suggested that the water balance remained normal. Therefore, the surrounding water was probably did not trigger the semi-aestivation in this teleost. Decrease of ammonia and increase of renal urea synthesis after 45 days of semi-aestivation led to the assumption that an alternative form of eliminating ammonia exists. Metabolic changes entailed by starvation were proposed to explain the biosynthesis of small molecules involved in the semi-aestivation of S. marmoratus.     

Stephanorhinus hundsheimensis (Mammalia,Rhinocerotidae) from the late early Pleistocene deposits of the Denizli Basin (Anatolia,Turkey)

Pleistocene rhinoceroses are poorly documented in Turkey where they have been reported only from the late early Pleistocene (1.3–1.1 Ma) travertine deposits of the Denizli Basin. In this work, new rhinoceros remains collected from this basin are assigned to a relatively large-sized Stephanorhinus hundsheimensis on the basis of their morphology and morphometry. The first Turkish record of this species is approximately coeval with the first appearance of S. hundsheimensis in Europe, chronologically referred to the late early Pleistocene, ca. 1.2 Ma. During that time, S. etruscus still survived in Iberian Peninsula, central Italy and Dacian Basin. The presence of two successive evolutionary morphs for S. hundsheimensis during the Pleistocene is not confirmed.     

New data on the historical and expanded range of Proterorhinus marmoratus (Pallas, 1814) (Teleostei: Gobiidae) in eastern Europe

A review of historical and recent area of distribution of tubenose goby Proterorhinus marmoratus was undertaken using critical analysis of published literature, data from two large fish collections (Natural History Museum, Kiev, Ukraine, and Zoological Institute, St Petersburg, Russia), personal collections of the authors sampled during extensive field activities in 1998–2004, and unpublished reports of the Institute of Lake and River Fisheries in Volgograd. Historically, riverine P. marmoratus from the Black Sea–Azov basin were distributed in the upper reaches of the rivers Dnieper and the Don even before the construction of water retention structures; this form at present has demonstrated only limited upstream range expansion. The native ‘Caspian tubenose goby’ is mostly connected with sea and delta habitats. We adduce some arguments for the tubenose goby's recent invasion of the Middle and Upper Volga as a result of introductions of the Don–Azov riverine form rather than upstream dispersal of the Caspian marine form.     

A new species of Youngomyia Felt from Brazil and new morphological data on Youngomyia pouteriae Maia (Insecta,Diptera, Cecidomyiidae)

Youngomyia matogrossensis Proença & Maia a new species of Cecidomyiidae (Insecta: Diptera) that induces cylindrical hairy galls on leaves of Pouteria torta (Mart.) Radlk. (Sapotaceae) is herein described and illustrated (larvae, pupal exuviae, male and female). The galler, gall and host plant were collected at Parque Nacional da Chapada dos Guimarães, in the state of Mato Grosso (Brazil). New morphological data and photographs of pupal exuviae, male and female of Youngomyia pouteriae Maia, 2001 are also provided.     

Brain-derived neurotrophic factor (BDNF)-like immunoreactivity localization in the retina and brain of Cichlasoma dimerus (Teleostei, Perciformes)

Brain-derived neurotrophic factor (BDNF) is a neurotrophin involved in the development and maintenance of vertebrate nervous systems. Although there were several studies in classical animal models, scarce information for fish was available. The main purpose of this study was to analyze the distribution of BDNF in the brain and retina of the cichlid fish Cichlasoma dimerus. By immunohistochemistry we detected BDNF-like immunoreactive cells in the cytoplasm and the nuclei of the ganglion cell layer and the inner nuclear layer of the retina. In the optic tectum, BDNF-like immunoreactivity was detected in the nucleus of neurons of the stratum periventriculare and the stratum marginale and in neurons of the intermediate layers. In the hypothalamus we found BDNF-like immunoreactivity mainly in the cytoplasm of the nucleus lateralis tuberis and the nucleus of the lateral recess. To confirm the nuclear and cytoplasm localization of BDNF we performed subcellular fractionation, followed by Western blot, detecting a 39 kDa immunoreactive-band corresponding to a possible precursor form of BDNF in both fractions. BDNF-like immunoreactivity was distributed in areas related with photoreception (retina), the integration center of retinal projections (optic tectum) and the control center of background and stress adaptation (hypothalamus). These results provide baseline anatomical information for future research about the role of neurotrophins in the adult fish central nervous system.     

Embryogenesis in mouth-breeding cichlids (Osteichthyes,Teleostei) structure and fate of the enveloping layer

The eggs of African mouth-brooders are of unusual size and shape. Studying their development may help to more clearly understand epiboly, gastrulation, and the relation between enveloping layer (periderm) and epidermis. When epiboly has progressed over just one fifth of the yolk mass, the germ ring and embryonic shield are already well established. Behind the germ ring very few deep cells are present at this early stage of epiboly, except in the embryonic shield. When the blastodisc covers the animal half of the yolk mass, the future body is already well established with notochord, somites and developing neural keel. Apart from these structures, no deep cells can be detected between enveloping layer and yolk surface; not even a germ ring remains behind the advancing edge of the enveloping layer. Epiboly over the greater part of the yolk is achieved only by the enveloping layer and the yolk syncytial layer. As the margin of the enveloping layer begins to reduce its circumference when closing around the vegetal pole, groups of cells in the advancing edge become spindle-shaped, with a single cell in between of each of these groups broadening along the edge. The enveloping layer (called periderm after epiboly) remains intact until after hatching, when, together with the underlying ectoderm, it forms the double-layered skin of the larval fish. Thereafter, cells deriving from the subperipheral ectoderm gradually replace the decaying periderm cells to form the final epidermis. Thus, in the cichlids studied, the enveloping layer alone forms the yolk sac to begin with, and it covers the larval body until some days after hatching.     

Sexual differences in the increase of white muscle fibres in Argentine hake, Merluccius hubbsi, from the San Matias Gulf (Argentina)

Growth dynamics of white fibres from axial muscle has been investigated in Argentine hake, Merluccius hubbsi , discriminating between sexes for the first time. The frequency distributions of fibre diameters are remarkably different in both sexes at sizes between 42 and 63·9 cm total length (T.l.). Males have a much lower proportion of newly recruited fibres (0–10 μm) than females; at 52–53·9 cm T.l., females have 4% of fibres in that category and males 0·5%. It appears that, from this size interval onwards, the increase in muscle mass is due only to the increase in diameter of individual fibres, which may exceed 300 μm. The lower recruitment rate of new fibres in males, and its relationship to lower growth rates and smaller final sizes, are discussed, and possible effects of reproductive activity are considered.     

Ultrastructure of Myxobolus brycon n. sp. (Phylum Myxozoa), parasite of the piraputanga fish Brycon hilarii (Teleostei) from Pantanal (Brazil)

Light and electron microscopy studies of a myxosporean, parasitizing the gill filaments of the freshwater fish Brycon hilarii (Valenciennes, 1850) (Characidae) collected in the Paraguay River (18°49'S, 57°39'W) (Pantanal), Brazil, is described. This parasite produces spherical to ellipsoidal polysporic histozoic plasmodia (Pmd) (up to ～180 μm in diameter) delimited by a double membrane and with several pinocytic channels. The plasmodial cyst contained the youngest developmental stages at the cortical periphery and immature and mature spores more internally. The Pmd developed near the cartilaginous structure of the gill filament, forming a prominent deformation where the gill lamellae disappear. Pyriform spores measured 6.9±0.6 (range 6.5-7.2) μm long, 4.2±0.5 (range 3.9-4.8) μm wide, and 2.5±0.7 (range 1.9-2.8) μm thick. The spores composed of two equal shell valves (～70 nm thick), adhering together along the straight suture line, surrounded two equal symmetric and elongated to pyriform polar capsules (PC) 4.2±0.6 (range 3.8-4.7) × 1.9±0.6 (1.7-2.5) μm; each PC contained a coiled polar filament with eight or nine (rarely 10) turns and a binucleated sporoplasm cell. Dense irregular masses were observed among the polar filaments coils. An intercapsular appendix was not observed. The sporoplasm contained several globular sporoplasmosomes randomly distributed among an extensive rough endoplasmic reticulum system with numerous vesicles and cisternae. Based on the morphological and ultrastructural differences and specificity of the host, we establish the new species, Myxobolus brycon n. sp.     

A new marine gobiid species of the genus Clariger Jordan & Snyder (Gobiidae, Teleostei) from Taiwan

A new species of Clariger Jordan & Snyder, 1901 was collected from northern Taiwan. The genus was previously known only from Japanese waters. This discovery is the first formal and southernmost record of these marine gobies from the waters of subtropical Taiwan. The new species, Clariger taiwanensissp. n., is distinguished from its congeners by a unique combination of features: (1) fin rays: dorsal-fin rays III, I/8; anal-fin rays modally I/8; and pectoral-fin rays modally 19 (2+16+1); (2) longitudinal dermal ridge on head with 6 barbels; and (3) specific coloration pattern: head and trunk dark brown with scattered pale spots and blotches; cheek, ventral portion of head sometimes pale with deep brown spots; pectoral-fin base with a dark brown band; and caudal fin mostly dark brown proximally and with alternating and irregular dark brown and pale bands distally. A diagnostic key to all nominal species from Japan and Taiwan is provided.     

Chromosomes, RAPDs and evolutionary trends of the Neotropical fish Mimagoniates microlepis (Teleostei: Characidae: Glandulocaudinae) from coastal and continental regions of the Atlantic forest, Southern Brazil

Chromosome and random amplified polymorphic DNA (RAPD) markers of samples of Mimagoniates microlepis were studied to test the hypothesis that a vicariant event occurred as the result of the orogeny of the coastal mountain range (Serra do Mar; southeastern and southern Brazil). Conventional karyotypes and nucleolar organizer region (Ag-NOR) phenotypes of two samples of M. microlepis from the headwaters of the Iguaçu River (southern Brazil) were compared both with each other and with other local populations of the species in the coastal drainage of southeastern Brazil. Additional molecular data (RAPD markers and genetic diversity) were obtained from specimens from coastal and continental regions of southern Brazil. The same diploid number (52 chromosomes), karyotypic formula and Ag-NOR phenotype were found for both analysed samples from the Iguaçu River. A genetic discontinuity was discovered in the comparison of the karyotypical formula of the Iguaçu samples with those from coastal drainages of the region. Polymerase chain reaction-RAPD markers revealed strikingly different molecular profiles between coastal and continental samples and indications of a high degree of genetic variation. Based on these results, we provide some comments on the biogeographical patterns and evolutionary trends for M. microlepis from coastal and continental regions of southeastern/southern Brazil.     

Monophyly, phylogenetic position and inter-familial relationships of the Alepocephaliformes (Teleostei) based on whole mitogenome sequences

Recent mitogenomic studies suggest a new position for the deep-sea fishes of the order Alepocephaliformes, placing them within the Otocephala in contrast to their traditional placement within the Euteleostei. However, these studies included only two alepocephaliform taxa and left several questions unsolved about their systematics. Here we use whole mitogenome sequences to reconstruct phylogenetic relationships for 11 alepocephaliform taxa, sampled from all five nominal families, and a large selection of non-alepocephaliform teleosts, to address the following three questions: (1) is the Alepocephaliformes monophyletic, (2) what is its phylogenetic position within the Teleostei and (3) what are the relationships among the alepocephaliform families? Our character sets, including unambiguously aligned, concatenated mitogenome sequences that we have divided into four (first and second codon positions, tRNA genes, and rRNA genes) or five partitions (same as before plus the transversions at third codon positions, using "RY" coding), were analyzed by the partitioned maximum likelihood and Bayesian methods. Our result strongly supported the monophyly of the Alepocephaliformes and its close relationship to the Clupeiformes and Ostariophysi. Altogether, these three groups comprise the Otocephala. Statistical comparison using likelihood-based SH test confidently rejected the monophyly of the Euteleostei when including the Alepocephaliformes. However, increasing the taxonomic sampling within the Alepocephaliformes did not resolve its position relative to the Clupeiformes and Ostariophysi. Within the Alepocephaliformes, our results strongly supported the monophyly of the platytroctid genera but not that of the remaining taxa. From one analysis to other, platytroctids were either the sister group of the remaining taxa or nested within the alepocephalids. Inferred relationships among alepocephaliform taxa were not congruent with any of the previously published phylogenetic hypotheses based on morphological characters.     

A prime inference on genetic diversity (RAPDs) in the marine fish Atherinella brasiliensis (Teleostei,Atherinopsidae) from Southern Brazil

Da Silva Cortinhas, M. C., Glienke, C., Prioli, A. J., Noleto, R. B., Matoso, D. A. and Cestari, M. M. 2010. A prime inference on genetic diversity (RAPDs) in the marine fish Atherinella brasiliensis (Teleostei, Atherinopsidae) from Southern Brazil. —Acta Zoologica (Stockholm) 91 : 242–248 As a result of the importance of Atherinella brasiliensis in estuarine environments, random amplified polymorphic DNA (RAPD) markers were used to verify the genetic diversity in A. brasiliensis from two different places in Paranaguá Bay (Paraná State) and one from the Conceição Lagoon (Santa Catarina State). Cytogenetic data have shown a high karyotypic diversity in some populations, although in others this peculiarity demonstrates rearrangements such as heterochromatinization. In the present study, a low level of genetic structuring between the samples from Conceição Lagoon compared with the others was observed through principal coordinate analysis (PCO), analysis of molecular variance and Mantel test according to 79 RAPD markers. As this specie does not perform horizontal migration and the individuals of Conceição Lagoon are isolated, three hypotheses are proposed to explain the results: (i) similar environments may show homogeneous populations not depending on the geographical distance, (ii) because vicariant events that formed the bays occurred in a recent period, the fragmentation effects over the structuring of the genetic diversity may still be low and not totally detectable by the RAPD technique and (iii) the isolation time or the number of generations may not be enough to promote a possible differentiation and genetic structuring between the specimens of these three places. The specimens of these places present a low level of differentiation and genetic structuring so we can consider them as a unique homogeneous population.     

A new species of Aphis Linnaeus from Guangxi,China (Hemiptera,Aphididae), based on morphological characters and DNA sequences

A new species of the genus Aphis with stridulatory apparatus, Aphis gnetuma Qiao sp. n., feeding on Gnetum parvifolium which is a new host plant record for aphids from Guangxi, China is described. The new species is distinguished by a well-developed median frontal tubercle and body bearing white wax powder in life. The taxonomic position of this new species is supported by analysis results of COI and gnd genes. A key to Aphis species with stridulatory spines on the hind tibiae of apterous viviparous females is provided. The type specimens studied are deposited in the National Zoological Museum of China, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.www.zoobank.org/urn:lsid:zoobank.org:act:BBB024DA-8454-4E22-8736-66ED92A3091B.     

The effects of testosterone implants on ovarian morphology in the bluehead wrasse, Thalassoma bifasciatum (Bloch) (Teleostei: Labridae)

The effects of exogenous testosterone pellet implants at 21 and 40 days on the ovary of Thalassoma bifasciatum , a protogynous marine fish, are described. The characteristic markings of the terminal blue phase appeared by 4 to 5 days and were complete by 18 days. There was no shift in sex ratio in the treated fish when compared to an untreated control group. None of the ovaries showed signs of precocious transformation, i.e., spermatogenic tissue was lacking, and there was no evidence of duct formation. Instead, however, the ovaries of the treated fish showed marked degenerative changes characterized by oocyte breakdown, fat infiltration, vacuolization, the accumulation of fibrous PAS+ material and the appearance of small eosinophilic cells.
These findings conflict with earlier studies on the same organism that employed different procedures. However, they resemble more closely the results obtained from work on another protogynoid, the ricefield eel, Monopterus albus. They add new light on the role of sex hormones in the reversal process of T. bifasciatum and hermaphroditic fishes in general.