Estudo imunológico e histopatológico da infecção experimental por Schistosoma mansoni em camundongos geneticamente selecionados para tolerância oral

A esquistossomose acomete 207 milhões de pessoas, com mais de 200 mil mortes anuais. Seu principal agente etiológico é o helminto Schistosoma e o principal modelo experimental, o camundongo. Linhagens de camundongos selecionadas geneticamente para susceptibilidade (TS) e resistência (TR) a tolerância imunológica constituem bons modelos para o estudo da resposta imunológica específica e inespecífica nas infecções. O objetivo deste trabalho foi caracterizar a infecção experimental por S. mansoni nestes camundongos, evidenciando a imunopatologia por diversos parâmetros na fase aguda da infecção. TR e TS não diferiram quanto a penetração de cercárias, recuperação de vermes adultos, fecundidade/produtividade de ovos das fêmeas de S. mansoni, mas predominaram ovos mortos em TS. Quanto maior o número de casais, maior a probabilidade de troca de casais e regressão sexual da fêmea, além de pequena redução da produtividade de ovos. Análise ultraestrutural dos parasitos machos recuperados de TS apresentaram tubérculos edemaciados, espinhos encurtados e em menor densidade que os parasitos dos TR. O tegumento dos parasitos recuperados de TS apresentou-se desorganizado, intensamente vacuolizado e com tendência a se desprender da superfície e espinhos internalizados e células vitelínicas desorganizadas. TS desenvolveram granulomas hepáticos grandes, com fibras radiais e predomínio do estágio exsudativo-produtivo com características de fase produtiva (EP/P), enquanto camundongos TR desenvolveram granulomas menores, com fibras concêntricas e predomínio de granulomas exsudativo-produtivos. TS desenvolveu hepatomegalia mais acentuada na fase aguda da infecção e exacerbada esplenomegalia na fase crônica. A aspartato aminotransferase mais elevada nos TR foi coerente com a acentuada histólise nos granulomas iniciais dos TR. É possível que a histólise menor em TS tenha contribuído para sua intensa hepatomegalia na fase aguda. Leucócitos totais séricos aumentaram em TS, nas fases aguda e crônica, mas não em TR. TS apresentaram anemia durante a fase crônica da infecção, possivelmente devido ao desvio na hematopoiese medular para a produção de leucócitos ou apoptose das hemácias. A mieloperoxidase neutrofílica hepática e no íleo foi maior em TS e a peroxidase de eosinófilos foi mais elevada no íleo do TS. Ambas as linhagens produziram IFN-γ, mas os níveis funcionais de IFN-γ foram diferentes nas duas linhagens em cultura de células. É possível que a imunopatologia hepática grave na linhagem TS possa estar relacionada aos altos títulos IFN-γ. TS produziu IL-10 em maior quantidade, entretanto esta citocina não foi capaz de regular o crescimento exacerbado dos granulomas hepáticos. Altos títulos de IL-4 na linhagem TS também são coerentes com a exacerbação dos granulomas, pois, como a IL-13, a IL-4 induz síntese de colágeno e está relacionada ao desenvolvimento da fibrose no granuloma esquistossomótico. Observamos redução do percentual relativo de células T CD4+ hepáticas de animais infectados em ambas as linhagens e redução percentual nas subpopulações de linfócitos B na medula óssea (precursores, linfócitos B imaturos, maduros e plasmócitos) mais acentuada em TS que em TR, possivelmente devido a extensa mobilização de B imaturos induzida pela inflamação ou desvio da hematopoiese para síntese de granulócitos em TS. Quantitativamente, TR não alterou suas subpopulações de linfócitos B. TS e TR são bons modelos para estudo da resposta imunológica na infecção esquistossomótica experimental. Novos estudos são necessários para confirmar nossas propostas e compreender os mecanismos envolvidos na diferença da resposta imunológica dessas linhagens na relação schistosoma-hospedeiro.

Larval development and identification of the genus Triglops (Scorpaeniformes: Cottidae)

Prior to Pietsch’s (1993) revision of the genus Triglops, identification of their larvae was difficult; six species co-occur in the eastern North Pacific Ocean and Bering Sea and three co-occur in the western North Atlantic Ocean. We examined larvae from collections of the Alaska Fisheries Science Center and Atlantic Reference Centre and used updated meristic data, pigment patterns, and morphological characters to identify larvae of Triglops forficatus, T. macellus, T. murrayi, T. nybelini, T. pingeli, and T. scepticus; larvae of T. metopias, T. dorothy, T. jordani, and T. xenostethus have yet to be identified and are thus not included in this paper. Larval Triglops are characterized by a high myomere count (42–54), heavy dorsolateral pigmentation on the gut, and a pointed snout. Among species co-occurring in the eastern North Pacific Ocean, T. forficatus, T. macellus, and T. pingeli larvae are distinguished from each other by meristic counts and presence or absence of a series of postanal ventral melanophores. Triglops scepticus is differentiated from other eastern North Pacific Ocean larvae by having 0–3 postanal ventral melanophores, a large eye, and a large body depth. Among species co-occurring in the western North Atlantic Ocean, T. murrayi and T. pingeli larvae are distinguished from each other by meristic counts (vertebrae, dorsal-fin rays, and anal-fin rays once formed), number of postanal ventral melanophores, and first appearance and size of head spines. Triglops nybelini is distinguished from T. murrayi and T. pingeli by a large eye, pigment on the lateral line and dorsal midline in flexion larvae, and a greater number of dorsal-fin rays and pectoral-fin rays once formed.

Age and growth of sailfish (Istiophorus platypterus) in waters off eastern Taiwan

Age and growth of sailfish (Istiophorus platypterus) in waters off eastern Taiwan were examined from counts of growth rings on cross sections of the fourth spine of the first dorsal fin. Length and weight data and the dorsal fin spines were collected monthly at the fishing port of Shinkang (southeast of Taiwan) from July 1998 to August 1999. In total, 1166 dorsal fins were collected, of which 1135 (97%) (699 males and 436 females) were aged successfully. Trends in the monthly mean marginal increment ratio indicated that growth rings are formed once a year. Two methods were used to back-calculate the length of presumed ages, and growth was described by using the standard von Bertalanffy growth function and the Richards function. The most reasonable and conservative description of growth assumes that length-at-age follows the Richards function and that the relationship between spine radius and lower jaw fork length (LJFL) follows a power function. Growth differed significantly between the sexes; females grew faster and reached larger sizes than did males. The maximum sizes in our sample were 232 cm LJFL for female and 221 cm LJFL for male.

Some aspects of the biology of Trypauchen vagina Bloch and Schneider of Bombay coast

Morphometrics of 14 metric and 6 meristic characters of Trypauchen vagina of Bombay coast is presented. Number of spines and rays of dorsal, anal and pectoral fins differ with earlier studies. Sexual dimorphism in the length of caudal and anal fins was observed. Length weight relationship of males and females have been worked out to be W = 0.00000 687L super(2.8440) and W = 0.00000 2747L super(3.0274) respectively. The fish is a prolific breeder and peak breeding activity occurs in December. The growth rate was found to be 15.20 mm/month.

Developmental characters of Psettina Iijimae (Jordan and Starks), bothid flat fishes - Pisces

Post larval stages of Psettina Iijimae ranging from 1.8 mm NL to 44.6 mm SL collected during Naga Expedition and International Indian Ocean Expedition (IIOE) are described. The characteristics which help to identify larval stages of Psettina are: the presence of pigmented urohyal appendage in early stages which is progressively reduced during flexion stages and which disappears in later postflexion stages, the meristics, the spines on urohyal and posterior basipterygial processes and the absence of spines on cleithra. The P. iijimae can be distinguished by the presence of spines on the median fin rays which differentiate near the baseosts along the dorsal and ventral body wall much before the fin rays. The larvae of P.iijimae were more abundant in the Gulf of Thailand compared to South China Sea and Indian Ocean.

Anchorage of air bladder with the interspinous bone of the anal fin in Lactarius lactarius (Bloch and Schneider, 1801)

Specimens of the false trevally (Lactarius lactarius ), 127 to 221 mm in total length, were studied for the mode of anchorage of the air-bladder with the interspinous bone of the anal fin. The 1st and 2nd interspinous bones are fused together into a single piece (named here as the anchor bone) which pierces through the air-bladder, dividing it into two intercommunicating chambers at its upper end, and ultimately articulates with the 10th vertebral bone. The lower end of the bone is broad, fan like with one side affording articulation with the 1st and 2nd anal spines. This is an unique feature of great taxonomical importance to L. lactarius, the only species in the family Lactariidae. The anal fin counts (23-27) and vertebral counts (23) are also given.

Larval ontogeny of a percichthyid fish, Synagrops philippinensis (Günther) in Kagoshima Bay, southern Japan

The larval ontogeny of a developmental series (1.2-8.3mm body length, BL) of Synagrops philippinensis from Kagoshima Bay, southern Japan is described and illustrated. The yolk was completely absorbed in larva of ≥1.5 mm BL. Notochord flexion commenced at about 3.5mm BL and was completed by about 4.0-4.5mm BL. S. philippinensis larvae were distinguished from their congeners based on melanophore patterns, head spination and fin spines and rays. Larvae of 7.5-8.3 mm BL were characterized by anteriorly serrated pelvic spine, two anal spines, nine inner preopercular spines and no melanophore on lateral side of the caudal peduncle; 7.0 to 7.5mm BL larvae by the above characters except serration on pelvic spine; and yolk-sac, pre-flexion, flexing and post-flexion larvae up to 7.0mm BL by unique melanophores on lower lobe of pectoral finfold/fin.

Larval development of a mangrove crab (Perisesarma bidens De Haan) (Crustacea: Brachyura: Sesarminae)

Larval development of Perisesarma bidens (De Haan) was investigated in laboratory conditions. Morphology of all larval stages and 1st crab stage was described and illustrated in detail, and compared with other species of sesarmid crabs. The zoeae morphological features of P. bidens are almost similar to other species of Sesarma in lacking a pair of lateral spines on carapace.

Early developmental stages of two Secutor species (Family: Leiognathidae) collected from the Bak-khali river estuary of the Bay of Bengal, Bangladesh

Early developmental stages of two Secutor species, Secutor insidiator (Bloch) (11.9-36.0 mm standard length, SL) and Secutor ruconius (Hamilton-Buchanan) (14.0-33.0 mm SL) collected by ichthyoplankton net from the Bak-khali river estuary of the Bay of Bengal, Cox's Bazar, Bangladesh are described and illustrated. All of the fins with supporting spines and rays were present in the smallest collected sizes of both species. With growth of the specimens, significant changes in melanophore patterns were found. S. insidiator is similar to S. ruconius in having upward protracting mouth parts and body colouration, but can be distinguished easily by its more elongate body shape (body depth 38-47% of SL compared with 46-52% of SL in Secutor ruconius). Both the species occurred round the year from August 1998 to July 1999. The surface water temperature and salinity during the study period varied from 22.0-32) C and 10-37 ppt respectively.

A new method for venom extraction from venomous fish, green scat

Scatophagus argus argus (Green Scat) is a pretty aquarium fish. Its hard spines are venomous and can cause painful injury. In this study 60 specimens of Green Scat were collected periodically from coastal waters of Boushehr (south of Iran) from May 2011 to April 2012. Anatomical features of venomous spines were investigated. Scat venom was extracted from the spines in a new manner for keeping the specimens alive. The nature of venom was tested by SDS-PAGE. Ethical issues and animal welfare principles such as rapid and instantaneous anesthetizing, post operation disinfection and fast recovery of the specimens was practiced in order to minimize the complications. This method enhanced the purity and quantity of venom as demonstrated by 12 separated proteins in electrophoresis. New ethical issues were developed to surviving the specimens and prolong viability as well.

Ecology and life history of penaeid shrimps

Fourteen species of penaeid shrimps with commercial value in Batan Bay and Tigbauan-Guimbal waters were identified as follows: Penaeus monodon, P.semisulcatus, P.japonicus, Metapenaeus ensis, M.burkenroadi, M.endeavouri, Metapenaeopsis palmensis, M.stridulans, Trachypenaeus fulvus, and Parapenaeus longipes. Among the 14 penaeids, P.semisulcatus, M.ensis and M.palmensis were found to be the dominant species within each genus. There are seven existing fishing gears for shrimping in the Batan Bay and Tigbauan-Guimbal waters: fish corrals, lift net, filter net, gill net, skimming net, baby trawler and commercial trawler. In general, female penaeids are larger than males. The largest P.monodon female measured was 81 mm in carapace length with 23 g in body weight. The largest male measuring 59 mm in carapace length with 119 g of body weight was caught in Batan Bay. Judging from spermatozoa occurrence on both sexes of P.monodon, the biological minimum size for male is 37 mm in carapace length and 49 mm for female. A total of 133 Penaeus postlarvae obtained from the offshore were identified by comparison with those reared in the laboratory. The postlarvae of P.japonicus-latisulcatus complex were quite dominant (60 . 2%), followed by P.semisulcatus (18 . 0%), and P.merguiensis-indicus complex (17 . 3%). The number of P.monodon postlarvae was relatively small (4 . 5%). The modal carapace length of P.monodon postlarvae from the offshore was 1 . 3 mm with three or four dorsal and no ventral spines on the rostrum, while P.monodon fry from the shoreline had 2 . 3 mm with five or six dorsal and one or two ventral spines.

Venomics and biological analysis of Scatophagus argus argus (Spotted scat) venom isolated in Persian Gulf, extraction and purification of hemolytic protein

Green scat namely as Scatophagus argus argus is a venomous aquarium fish belonging to Scatophagidae family. It can induce painful wounds in injured hand with partial paralysis to whom that touch the spines. Dorsal and ventral rough spines contain cells that produce venom with toxic activities. According to unpublished data collected from local hospitals in southern coastal region of Iran, S. argus is reported as a venomous fish. Envenomation induces clinical symptoms such as local pain, partial paralysis, erythema and itching. In the present study green scat (spotted scat) was collected from Persian Gulf coastal waters. SDS-PAGE indicated 12 distinct bands in the venom ranged between 10-250 KDa. The crude venom had hemolytic activity on human erythrocytes (1%) with an LC100 (Lytic Concentration) of about 1.7 μg. The crude venom can release 813 μg proteins from 0.5% casein. Phospholipase C activity was recorded at 3.125 μg of total venom. Our findings showed that the edematic activity remained over 48 h after injection. The purification of the venom was done by HPLC and 30 peaks were obtained within 80 min but only one peak in 68 min retention time showed hemolytic activity at 90% acetonitril was isolated. The area percentage of the hemolytic protein showed that this hemolytic protein consist of 32 percent of total proteins and its molecular weight was 72 KDa in SDS_PAGE. The results demonstrated that crude venom extracted from Iranian coastal border has different toxic and enzymatic activities.

Phylogenetic relationships of the mustache toads inferred from mtDNA sequences

Species of the genus Vibrissaphora are unique among all annurans in that males exhibit external cornified spines in the maxillary region during the breeding season. They were separated from species of the genus Leptobrachium based on this unique character. We construct a phylogeny using the 16S, ND4, and cytochrome b mitochondrial genes of 42 individuals from eight species of Vibrissaphora and five species of Leptobrachium from mainland China, Southeast Asia, and Hainan Island. Species of both Oreolalax and Scutiger were used as outgroups. The results indicate that: L. huashen and L. chapaense form a clade that is nested within Vibrissaphora, and L. hainanense is the sister taxon to the clade comprising all Vibrissaphora plus L. chapaense and L. huashen; V. boringiae is grouped with a clade consisting of V. leishanensis, V. liui, and V. yaoshanensis; and V. yaoshanensis is a species separate from V. liui. We propsed taxonomix changes that reflect these findings. Also based on the resulting phylogenetic trees, we propose that the mustache toads originated in the trans-Himalayan region of southwest China, and that the evolution of maxillary spines, large body size, and reverse sexual size dimorphism in these frogs was influenced by intrasexual selection due to adopting a resource-defense polygyny matting system.

MtDNA phylogeny provides evidence of generic polyphyleticism for East Asian bagrid catfishes

We explore the intrafamilial relationships of East Asian bagrid catfishes (Hemibagrus, Pseudobagrus, Pelteobagrus, and Leiocassis) based on 245 sequences of 1092 bp mitochondrial cytochrome b fragments. Four haplotypes were found to be shared by Pseudobagrus ussuriensis, Pelteobagrus vachelli and Pelteobagrus nitidus. Phylogenetic trees were performed using the neighbor joining, maximum parsimony, maximum likelihood, and Bayesian likelihood methods. The phylogenetic trees based on NJ, MP, ML and BL inferences strongly support polyphyleticism for the currently recognized genera Pseudobagrus, Pelteobagrus and Leiocassis. However, the species currently assigned to these three genera form a robustly monophyletic group with relatively low genetic divergence. The structure of maxillary barbels and serrations on the anterior edge of the pectoral spines seem to be indicatory of appropriate phylogenetic traits. We propose that only Hemibagrus and Pseudobagrus are the only valid genera of East Asian bagrids.

Age and growth of an endemic cyprinid fish, Spinibarbus yunnanensis, from Lake Fuxian, China

Age and growth of Spinibarbus yunnanensis Tsu in Lake Fuxian were investigated by examination of annuli from scales, dorsal fin spines, and otoliths. Sectioned otoliths exhibited the clearest and the most regular annuli. The formation of false annuli, which regularly arose on scales and dorsal fin spines during the juvenile stage, may be caused by a change in diet during development. The parameters for the von Bertalanffy growth curve were: K = 0.105, L-infinity = 950 mm, t(0) = -0.22 year for both sexes. W-infinity, calculated from the relationship W-infinity = aL(infinity)(b), was 10,352 g.