Proterodiplostome Parasites (Digenea, Proterodiplostomidae) of the Caiman, Caiman crocodilus yacare (Reptilia, Crocodylia) in the Pantanal mato-Grossense, Brazil, with the description of two new species

Two new species are described from the caiman. Caiman crocodilus yacare. Proterodiplostomum breve n. sp. differs from all other species in the genus by the following chacacteristics: (1) the paraprostate gland is shorter and club-shaped; (2) the genital cone is, in average, eight times longer than that of P. medusae; (3) the genital atrium is larger and without pseudosuckers; (4) the oral sucker and pharynx are longer; and (5) there are larger numbers of papillae surrounding the tribocytic organ (40) against 20 in P. longum, 16 in p. tumidilum, 8 in P. ophidum, and 16-18 in P. medusae. Proterodiplostomum globulare n. sp. differs from all the other species in the fenus by the following characteristics: (1) from P. tumidilum, P. lomgum, P. medusae, and P. breve n. sp. for the absense of pseudosuckers or muscular bunches in the inferior wall of the genital atrium; (2) the shape of the paraprostate gland, which is globular and not cylindrical as in P. longum, P. tumidilum, P. medusae, and P. ophidum; (3) the size of the tribocytic organ 201-407 long, 183-495 wide, while is 138-270 long, 102-292 wide in P. medusae, and 138-270 long, 255 wide in P. ophidum; (4) the number of papillae in the tribocytic organ (18-20) in P. globulare and 16-18 in P. medusae, and 8 in P. ophidum. Specimens belonging to six other species of proterodiplostomes are recorded for the first time infecting the caiman, C. c. yacare in the Pantanal Mato-grossense, Brazil, namely: Proterodiplostomum medusae, P. tumidilum, Cystodiplostomum hollyi, Prolecithodiplostomum constrictum, Paradiplostomum abbreviatum, and Herpetodiplostomum caimancola.

Eimeria peltocephali n. sp., (Apicomplexa:Eimeriidae) from the Freshwater Turtle Peltocephalus dumerilianus (Chelonia:Pelomusidae) and Eimeria molossi n. sp., from the Bat, Molossus ater (Mammalia:Chiroptera)

The oocyst is described of Eimeria peltocephali n.sp. from faeces of the freshwater turtle Peltocephalus dumerilianus from Barcelos, State of Amazonas, Brazil. Sporulation is exogenous and fully developed oocysts are elongate, ellipsoidal or cylindrical, frequently curved to a banana-shape, 54.4 x19.1 (37.5 - 68.7 x 18.7-20.0 µm), shape-index 2.8 (1.8 -3.9). The oocyst wall is a single thin, colourless layer about 1 µm thick, with no micropyle. There is a bulky oocyst residuum, at first spherical to ellipsoidal, 19 x 16 (16. 2 -26.2 x 16 - 21.5µm) , but becoming dispersed on maturation. There are no polar bodies. The sporocysts, 19.1 x 6.8 ( 17.5 -21.2 x 6.2 -7.5 µm), shape- index 2.8 (2.3 -3.2), are usually disposed in pairs at each end of the oocyst, and bear an inconspicuous Stieda body in the form of a flat cap. The sporozoites are elongate and slightly curved around the residuum. No refractile bodies were seen. Eimeria molossi n.sp., is described from the molossid bat Molossus ater. Sporulation is exogenous and the mature oocysts are predominantly broadly ellipsoidal, 23.4 x 17.5 (18-30 x 15-22.5 µm), shape-index 1.3 (1-1.6). The oocyst wall is about 2 µm thick, and of three layers: an inner thin, colourless one and two outer layers which are thicker, yellowish-brown, prominently striated and in close apposition. There is no micropyle or oocyst residuum, but one and occasionally two polar bodies are usually present. Sporocysts are ellipsoidal, 10.2 x 7.5 (10-12.5 x 7.5 µm), shape-index 1.4 (1.3-1.7) with an inconspicuous Stieda body. Endogenous stages are described in the epithelial cells of the small intestine

Ultrastructure of the membrane attack complex of complement: detection of the tetramolecular C9-polymerizing complex C5b-8.

The ultrastructure of the membrane attack complex (MAC) of complement had been described as representing a hollow cylinder of defined dimensions that is composed of the proteins C5b, C6, C7, C8, and C9. After the characteristic cylindrical structure was identified as polymerized C9 [poly(C9)], the question arose as to the ultrastructural identity and topology of the C9-polymerizing complex C5b-8. An electron microscopic analysis of isolated MAC revealed an asymmetry of individual complexes with respect to their length. Whereas the length of one boundary (+/- SEM) was always 16 +/- 1 nm, the length of the other varied between 16 and 32 nm. In contrast, poly(C9), formed spontaneously from isolated C9, had a uniform tubule length (+/- SEM) of 16 +/- 1 nm. On examination of MAC-phospholipid vesicle complexes, an elongated structure was detected that was closely associated with the poly(C9) tubule and that extended 16-18 nm beyond the torus of the tubule and 28-30 nm above the membrane surface. The width of this structure varied depending on its two-dimensional projection in the electron microscope. By using biotinyl C5b-6 in the formation of the MAC and avidin-coated colloidal gold particles for the ultrastructural analysis, this heretofore unrecognized subunit of the MAC could be identified as the tetramolecular C5b-8 complex. Identification also was achieved by using anti-C5 Fab-coated colloidal gold particles. A similar elongated structure of 25 nm length (above the surface of the membrane) was observed on single C5b-8-vesicle complexes. It is concluded that the C5b-8 complex, which catalyzes poly(C9) formation, constitutes a structure of discrete morphology that remains as such identifiable in the fully assembled MAC, in which it is closely associated with the poly(C9) tubule.

Description of Eimeria motelo sp. n. (Apicomplexa: Eimeriidae) from the yellow footed tortoise, Geochelone denticulata (Chelonia: Testudinidae), and replacement of Eimeria carinii Lainson, Costa & Shaw, 1990 by Eimeria lainsoni nom. nov.

Eimeria motelo sp. n. is described from faeces of the yellow-footed tortoise, Geochelone denticulata (L.). Oocysts are irregularly ellipsoidal or cylindrical, with slightly expressed lobed protrusions and irregularities at the poles, possibly caused by wrinkling of the oocyst wall, 17 (15-19) × 9.4 (8.5-11) µm, shape index (length/width) being 1.81 (1.45-2). The oocyst wall is smooth, single-layered, 0.5 µm thick with no micropyle. There are no polar bodies. Sporocysts are ellipsoidal, 8.9 (7.5-10) × 4.4 (4-5) µm, shape index 2.03 (1.7-2.5). A sporocyst residuum is present, composed of many granules of irregular size. The sporozoites are elongate, lying lengthwise in the sporocysts. Comparison with other species of the genus Eimeria parasitising members of family Testudinidae indicates that the presently described coccidium represents a new species. The name of Eimeria carinii Lainson, Costa & Shaw, 1990 is found to be preoccupied by a homonym, Eimeria carinii Pinto 1928 given to a coccidium from Rattus norvegicus. Therefore, it is replaced by Eimeria lainsoni nom. nov.

Avaluació de la teratogènia in vitro mitjançant el cultiu d’embrions sencers (WEC)

Projecte de recerca elaborat a partir d’una estada a la Charité - Universitätsmedizin Berlin, Alemanya, entre novembre i desembre del 2007. En aquest treball es presenta el protocol a seguir per a dur a terme el cultiu d’embrions sencers in vitro (Whole Embryo Culture, WEC). Amb aquest protocol es pretén implementar la tècnica del WEC en el laboratori de la Unitat de Toxicologia de la Facultat de Farmàca (UB), seguint la metodologia apresa durant l’estada i deixant per escrit tots els passos seguits i el material i la metodologia concreta de cadascun d’ells. En el WEC es cultiven embrions de rata de 9.5 dies durant 48h en ampolles rotatòries en un medi líquid i amb una fase gasosa controlats. Durant el cultiu, tenen lloc dos processos principals: el plegament de l’embrió i l’organogènesi. Els embrions durant els dos dies que dura el cultiu es pleguen en els plans transversal i sagital, passant d’un embrió pla a un altre de cilíndric en forma de “C”. En aquest període, a més, es produeixen importants processos d’organogènesi com la neurulació, la formació de la cresta neural, dels somites, dels vasos sanguinis - el cor inclòs- i de la sang. Es comencen a formar la placoda nasal, la vesícula oftàlmica, la vesícula òtica, les extremitats superiors i inferiors i la cua. En la memòria adjunta es descriuen amb detall els processos d'aparellament dels animals, preparació del material i del medi de cultiu, el procés d'aïllament del embrions en el dia 9.5, les condicions de cultiu i l'avaluació dels embrions en el dia 11.5. Finalment es presenten resultats d'embrions en situació control amb un correcte desenvolupament i es mostra com, al final de l'estada, es va aconseguir el cultiu d’embrions control amb un desenvolupament correcte i estadísticament sense diferències respecte als diferents paràmetres mesurats en comparació amb els embrions control de la Charité-Universitätsmedizin de Berlin.

Description of immature stages of Culex ocossa Dyar & Knab, Culex delpontei Duret and Culex pereyrai Duret of the Melanoconion subgenus (Diptera: Culicidae)

The larva and pupa of Culex (Melanoconion) ocossa Dyar & Knab are redescribed and those of Culex (Melanoconion) delpontei Duret and Culex (Melanoconion) pereyrai Duret are described from specimens collected in the states of São Paulo and Paraná, Brazil. The pupa of Cx. ocossa differs from those of the other two species in having seta 5-IV-VI dark with strongly aciculated branches, and caudolateral angle of segment VIII produced into sharp point, and seta 1-P present; Cx. delpontei can be distinguished from Cx. pereyrai in possessing paddle lightly tanned, trumpet flared, and wing and leg cases lightly tanned, without pattern of dark spots; Cx. pereyrai can be recognized by having wing case with pattern of dark, discontinuously pigmented, longitudinal lines, and trumpet cylindrical, not flared. The larvae of the three species share the presence of seta 2-C placed medially to seta 1-C.

Gregarine Cephaloidophora communis mawrodiadi, 1908 in the barnacle Euraphia rhyzophorae, Oliveira, 1940 from Brazil

The gregarine Cephaloidophora communis was observed for the first time in Brazil in the barnacles Euraphia rhyzophorae collected in Angra dos Reis, Rio de Janeiro, Brazil, between 1990 and 1996. Histological studies showed growth phases of the parasite in specific parts of the digestive system. The intracellular forms occurred in the vacuoles of the intestinal cells. Syzygy was frequent, and the most common form following syzygy was cylindrical, with a single membrane. The cytoplasm of the gregarines was always irregular, dense, and occasionally presenting a dark stoch area.

Aquàriums i educació : una evolució paral·lela

Com es pot fer que la visita a una instal·lació museogràfica sigui informativa, lúdica i a la vegada instructiva? Aquest treball es proposa esbrinar quines orientacions museogràfiques amb finalitat educativa s'han portat a terme en els aquàriums, contextualitzades amb una perspectiva històrica i exemplaritzades en una instal·lació concreta, que és L'Aquàrium de Barcelona.

Some coccidial parasites of the lizard Amphisbaena alba (Reptilia: Amphisbaenia: Amphisbaenidae)

Five parasites are described in the lizard Amphisbaena alba (Amphisbaenidae) from the state of Pará, North Brazil. Mature oocysts of Choleoeimeria amphisbaenae n. sp., are passed already mature in the faeces. They are ellipsoidal-cylindrical, average 33.7 x 22.8 µm and are devoid of micropyle, oocyst residuum or polar body. The colourless wall is smooth and of 2 layers. The 4 dizoic sporocysts have no Stieda body and average 13 x 9.3 µm. Endogenous stages develop in the epithelial cells of the gall-bladder in the manner described for the genus and may cause extensive tissue damage. Sporulation of Isospora capanemaensis n. sp., is completed 3 days after the oocysts are voided in the faeces. They average 14.8 x 14.5 µm and have no micropyle, oocyst residuum or polar body. The 2 tetrazoic sporocysts are pear-shaped, average 8.6 x 6.6 and have an inconspicuous Stieda body. Endogenous development is in the epithelial cells of the ileum, and heavy infections cause considerable tissue destruction. Multisporocystic oocysts passed in the faeces of one A. alba possibly originated from an invertebrate host ingested by the lizard. A globidium-like cyst in the digestive tract of A. alba measured 105 x 85 µm and contained many hundreds of merozoites. A stained kidney smear of the same lizard revealed the presence of an unidentified parasite producing multinucleate cyst-like stages.

Diagnostic of Biomphalaria snails and Schistosoma mansoni: DNA obtained from traces of shell organic materials

Freshwater snails belonging to the genus Biomphalaria act as intermediate hosts for the parasite trematode Schistosoma mansoni in Africa and in the neotropical region. Identification of such molluscs is carried out based on morphological characters and the presence of cercariae is verified through squeezing snails between two glass slides or by exposing them to artificial light. However, sometimes, the material collected includes molluscs with decomposed bodies or, yet, only empty shells, which precludes their identification and S. mansoni detection. Due to these difficulties, we have developed a methodology in which DNA may be extracted from traces of organic material from inside shells in order to identify molluscs through polymerase chain reaction and restriction fragment length polymorphism and to detect S. mansoni into these snails, by using low stringency polymerase chain reaction. Species-specific profiles obtained from B. glabrata, B. straminea, and B. tenagophila snails and their shells, maintained in laboratory for ten years, showed the same profiles. S. mansoni profiles showed to be present in shell specimens as far as the eighth week after being removed from aquarium.

Evaluation of Mesocyclops annulatus (Copepoda: Cyclopoidea) as a control agent of Aedes aegypti (Diptera: Culicidae) in Argentina

We evaluated the potential of Mesocyclops annulatus as a control agent of Aedes aegypti in La Plata city (Argentina). Mosquito larval survivorship due to predation by these copepods was estimated at weekly intervals during the oviposition period of A. aegypti. Mean weekly A. aegypti larval survivorship in cylindrical plastic containers (12 cm height and 11 cm diameter) with copepods was significantly lower than in control containers. Furthermore, weekly larval survival was negatively correlated with M. annulatus adult density, and approximately 23 adult copepods/container would be a threshold density over which the weekly mosquito larval survivorship approached zero. The copepods were able to persist in all containers during approximately 100 days (in three of them until the end of the experiment: 155 days) without the resource represented by A. aegypti larvae. The predation and persistence observed suggest that M. annulatus is a potential control agent to be considered in biological control programs.

Standardisation of 18F by a coincidence method using full solid angle detectors.

A solution of (18)F was standardised with a 4pibeta-4pigamma coincidence counting system in which the beta detector is a one-inch diameter cylindrical UPS89 plastic scintillator, positioned at the bottom of a well-type 5''x5'' NaI(Tl) gamma-ray detector. Almost full detection efficiency-which was varied downwards electronically-was achieved in the beta-channel. Aliquots of this (18)F solution were also measured using 4pigamma NaI(Tl) integral counting and Monte Carlo calculated efficiencies as well as the CIEMAT-NIST method. Secondary measurements of the same solution were also performed with an IG11 ionisation chamber whose equivalent activity is traceable to the Système International de Référence through the contribution IRA-METAS made to it in 2001; IRA's degree of equivalence was found to be close to the key comparison reference value (KCRV). The (18)F activity predicted by this coincidence system agrees closely with the ionisation chamber measurement and is compatible within one standard deviation of the other primary measurements. This work demonstrates that our new coincidence system can standardise short-lived radionuclides used in nuclear medicine.

Lanfrediella amphicirrus gen. nov. sp. nov. Nematotaeniidae (Cestoda: Cyclophyllidea), a tapeworm parasite of Rhinella marina (Linnaeus, 1758) (Amphibia: Bufonidae)

The family Nematotaeniidae, tapeworms commonly found in the small intestines of amphibians and reptiles, includes 27 recognised species distributed among four genera: Bitegmen Jones, Cylindrotaenia Jewell, Distoichometra Dickey and Nematotaenia Lühe. The taxonomy of these cestodes is poorly defined, due in part to the difficulties of observing many anatomical traits. This study presents and describes a new genus and species of nematotaeniid parasite found in cane toads (Rhinella marina) from eastern Brazilian Amazonia. The cestodes were collected during the necropsy of 20 hosts captured in the urban area of Belém, Pará. The specimens were fixed and processed for light microscopy, scanning electron microscopy (SEM) and three-dimensional (3D) reconstruction. Samples were also collected for molecular analyses. The specimens presented a cylindrical body, two testes and paruterine organs. However, they could not be allocated to any of the four existing nematotaeniid genera due to the presence of two each of dorsal compact medullary testes, cirri, cirrus pouches, genital pores, ovaries and vitelline glands per mature segment. Lanfrediella amphicirrus gen. nov. sp. nov. is the first nematotaeniid studied using Historesin analysis, SEM and 3D reconstruction, and it is the second taxon for which molecular data have been deposited in GenBank.

Real-time recording of circadian liver gene expression in freely moving mice reveals the phase-setting behavior of hepatocyte clocks.

The mammalian circadian timing system consists of a master pacemaker in the suprachiasmatic nucleus (SCN) in the hypothalamus, which is thought to set the phase of slave oscillators in virtually all body cells. However, due to the lack of appropriate in vivo recording technologies, it has been difficult to study how the SCN synchronizes oscillators in peripheral tissues. Here we describe the real-time recording of bioluminescence emitted by hepatocytes expressing circadian luciferase reporter genes in freely moving mice. The technology employs a device dubbed RT-Biolumicorder, which consists of a cylindrical cage with reflecting conical walls that channel photons toward a photomultiplier tube. The monitoring of circadian liver gene expression revealed that hepatocyte oscillators of SCN-lesioned mice synchronized more rapidly to feeding cycles than hepatocyte clocks of intact mice. Hence, the SCN uses signaling pathways that counteract those of feeding rhythms when their phase is in conflict with its own phase.

Culicidae (Diptera) selection of humans, chickens and rabbits in three different environments in the province of Chaco, Argentina

Studies were conducted to determine the selection of humans, chickens and rabbits by Culicidae in three different environments in the province of Chaco, Argentina. Mosquitoes were collected fortnightly using cylindrical metal traps containing animal bait (chickens and rabbits). The mosquitoes were collected between June 2001-May 2002. During the same period and with the same frequency, mosquitoes biting the human operators of the traps were collected during the first 15 min of exposure within different time intervals: from 09:00 am-11:00 am, 01:00 pm-03:00 pm, 05:00 pm-07:00 pm and 09:00 pm-10:00 pm. A total of 19,430 mosquitoes of 49 species belonging to 10 genera were collected. Culex species mainly selected chicken bait and Wyeomyia species selected rabbit bait. Ochlerotatus and Psorophora species were more abundant in rabbit-baited traps. Anopheles triannulatus, Coquillettidia nigricans, Ochlerotatus scapularis, Mansonia titillans and Psorophora albigenu showed a strong attraction for human bait. The Anopheles, Coquillettidia, Culex and Mansonia species were more active between 05:00 pm-09:00 pm, while Ochlerotatus, Psorophora, Haemagogus and Wyeomyia were most active from 09:00 am-07:00 pm. This study provides additional information about the biology and ecology of arbovirus vectors in Chaco.