Testing hypotheses on the origin and dispersion of <Emphasis Type="Italic">Limnoperna fortunei</Emphasis> (Bivalvia,Mytilidae) in the Iguassu River (Paraná, Brazil): molecular markers in larvae and adults

The invasive bivalve Limnoperna fortunei (Dunker 1857) was introduced in South America in 1991, with the first occurrence in Brazil in 1998. In the Iguassu River, the species was recorded in 2001; however, it is unknown how it was introduced and spread. Adults and larvae were sampled in Iguassu and Paraná Rivers, and the genetic profiles were compared. The species was absent in the upper reaches and only larvae were found, in low densities, in intermediary reaches. The L. fortunei populations from the lower Iguassu River presented no genetic differentiation among themselves, suggesting strong connectivity, and were significantly different from the Paraná River populations, most likely because of Iguassu Falls. Furthermore, the results suggest that the Paraná River represents the source of propagules to the Iguassu River. Generally, no significant differences were observed between the genetic structure inferred from adults and larvae. Only the population from the Iguassu National Park, a lotic environment, differed from the remaining subpopulations. The characterization of genetic profile using larval stages of L. fortunei populations was satisfactory, and represents an important protocol for studying the population genetics of aquatic species with planktonic larval stages.     

Neotropical Monogenoidea. 43. Diplectanum monticellii n. sp. (Diplectanidae) from the gills of Cynoscion leiarchus (Perciformes: Sciaenidae) in Brazil

Diplectanum monticellii n. sp. is described from the gills of Cynoscion leiarchus, a marine Sciaenidae, from Itacuru?á, Rio de Janeiro (type locality); Baía de Guaratuba, Paraná; and Pontal do Sul, Paraná in Brazil. The new species is characterized by the following features: bell-shaped male copulatory organ with a sleevelike base, accessory piece absent, vas deferens looping left intestinal cecum, vaginal aperture sinistroventral, and ventral anchor with elongate superficial and deep roots.     

Neotropical Monogenoidea. 50. Oviparous gyrodactylids from loricariid and pimelodid catfishes in Brazil, with the proposal of Phanerothecioides n. g., Onychogyrodactylus n. g. and Aglaiogyrodactylus n. g. (Polyonchoinea: Gyrodactylidea)

The diagnosis of Phanerothecium Kritsky & Thatcher, 1977 is amended, and Phanerothecioides n. g., Onychogyrodactylus n. g. and Aglaiogyrodactylus n. g., all comprised of oviparous gyrodactylids (Monogenoidea: Polyonchoinea), are proposed to accommodate 11 of the 15 species (14 new to science) herein described and/or reported from loricariid and pimelodid catfishes (Teleostei: Siluriformes) in Brazil: Phanerothecium harrisi Kritsky & Boeger, 1991, P. spinatoides n. sp. and P. deiropedeum n. sp. all from Hypostomus spp.; P. spinulatum n. sp. from Hypostomus cf. regani; Phanerothecioides agostinhoi n. g., n. sp. (type-species) from Hypostomus spp. and Pseudoplatystoma fasciatum; Onychogyrodactylus sudis n. g., n. sp. (type-species) and O. hydaticus n. sp. both from Ancistrus multispinus; Aglaiogyrodactylus forficulatus n. g., n. sp. (type-species) from Kronichthys lacerta; A. pedunculatus n. sp. from Hisonotus sp.; A. guttus n. sp. from Pseudotothyris obtusa; A. salebrosus n. sp., A. conei n. sp. and A. ctenistus n. sp. all from Pareiorhaphis parmula; and A. calamus n. sp. and A. forficuloides n. sp. both from Schizolecis guntheri. Phanerothecioides n. g. is characterised by oviparous forms lacking superficial and deep haptoral bars and pregermarial vitelline follicles, and by having a conspicuous testis in adult specimens, a syncytial prostatic gland, a reduced copulatory sac and vitelline ducts in the form of an inverted ‘U’. Onychogyrodactylus n. g. is differentiated from all other oviparous gyrodactylid genera by its members having a spine-like accessory sclerite enclosed in a separate pouch associated with the terminal male genitalia. Species of Aglaiogyrodactylus n. g. possess H-shaped vitelline ducts and a complex accessory piece and sclerotised or non-sclerotised male copulatory organ enclosed within the copulatory sac.     

Histopathology of the mangrove land crab Ucides cordatus (Ocypodidae) affected by lethargic crab disease

Lethargic crab disease (LCD) has caused extensive epizootic mortality of the mangrove land crab Ucides cordatus (Linnaeus 1763) (Brachyura: Ocypodidae) along the Brazilian coast. Direct culture of tissue samples from sick crabs and subsequent isolation and purification identified the causative agent as an Exophiala species of fungus. The histopathology of crabs with variable signs of LCD indicates that the most affected tissues are the epidermis, connective tissue, heart, hepatopancreas, nervous system, and gills. Gonads, somatic muscles, and digestive system are less affected by the fungus. The observed pathology is compatible with the clinical signs of LCD. Necrosis, tissue degeneration, and congestion of hemal sinuses and vessels are present in heavily infected organs. Nerve fibers may be compressed by accumulations of yeast-like cells. In heavy infections the tissue of gill lamellae is destroyed with subsequent dilation or compression. Cellular immune responses include hemocytic infiltration, agglutination and encapsulation, and phagocytosis. Phagocytosis of yeast-like cells is abundant in the connective tissue associated with the exoskeleton. These results indicate that LCD is the result of a systemic phaeohyphomycosis caused by a species of Exophiala. The present study also suggests that dispersal of the fungus within the crab occurs through the hemal system.     

Nonglutamate pore residues in ion selection and conduction in voltage-gated Ca(2+) channels

High-affinity, intrapore binding of Ca(2+) over competing ions is the essential feature in the ion selectivity mechanism of voltage-gated Ca(2+) channels. At the same time, several million Ca(2+) ions can travel each second through the pore of a single open Ca(2+) channel. How such high Ca(2+) flux is achieved in the face of tight Ca(2+) binding is a current area of inquiry, particularly from a structural point of view. The ion selectivity locus comprises four glutamate residues within the channel's pore. These glutamates make unequal contributions to Ca(2+) binding, underscoring a role for neighboring residues in pore function. By comparing two Ca(2+) channels (the L-type alpha(1C), and the non-L-type alpha(1A)) that differ in their pore properties but only differ at a single amino acid position near the selectivity locus, we have identified the amino-terminal neighbor of the glutamate residue in motif III as a determinant of pore function. This position is more important in the function of alpha(1C) channels than in alpha(1A) channels. For a systematic series of mutations at this pore position in alpha(1C), both unitary Ba(2+) conductance and Cd(2+) block of Ba(2+) current varied with residue volume. Pore mutations designed to make alpha(1C) more like alpha(1A) and vice versa revealed that relative selectivity for Ba(2+) over K(+) depended almost solely on pore sequence and not channel type. Analysis of thermodynamic mutant cycles indicates that the motif III neighbor normally interacts in a cooperative fashion with the locus, molding the functional behavior of the pore.     

拼接信号序列单碱基变异提高马传染性贫血病毒mRNA拼接效率

分别以马传染性贫血(马传贫)驴强毒(D—A EIAV)RNA和马传贫驴白细胞弱毒疫苗(DLA EIAV)RNA为模板,利用RT—PCR的方法,克隆到马传贫强、弱毒株基因组外显子2及其下游的核苷酸序列。然后将报告基因CAT插入到EIAV内含子2env阅读框架中,构成CAT拼接报告系统。同时在强毒株重组表达质粒的基础上,将其外显子-3上游拼接受体位点的核苷酸序列CAG突变为弱毒株相应位置的核苷酸序列TAG,得到强毒单核苷酸突变株重组表达质粒。用构建的3个重组表达质粒DNA转染驴血白细胞,ELISA检测转染细胞CAT浓度。结果表明：EIAV强毒株重组表达质粒中CAT蛋白表达量最高,EIAV强毒株重组表达质粒次之,EIAV强毒突变株重组表达质粒最低。由于CAT基因被插入于各重组质粒中的EIAV内含子-2里,EIAV外显子-2、3之间的拼接可导致该基因的删除,因而其拼接效率低于EIAVmRNA外显子-2、3之间的拼接效率。实验数据表明,EIAV SA2拼接信号序列单碱基变异提高了SD2-SA2拼接效率;D—AEIAV SA2-SD2拼接效率比DLA EIAV相应位点拼接效率高。     

结核分枝杆菌可视化抗体检测蛋白芯片的制备

目的：利用克隆表达的7种结核分枝杆菌优势表位抗原,建立可视化抗体检测蛋白芯片,用于结核病辅助诊断。方法：将7种结核分枝杆菌优势表位抗原,即38kD、ESAT-6、CFP10、MPT64、Mtb8、Mtb8.4和Mtb16.3点于修饰的基片上,制备可检测7种结核抗体的多靶点蛋白微阵列,建立免疫金银染色检测系统;使用该芯片对48例临床结核病患者血液样品进行检测,并与“金标准”痰涂片（48例）和痰培养（其中的29例）检测结果进行比较,分析其敏感性;对30名献血员血液样品进行检测,分析其特异性。结果：可视化抗体检测蛋白芯片的敏感性分别为98.5％和96.6％,而痰涂片和痰培养检测方法的敏感性分别为35.4％和48.3％;可视化抗体检测蛋白芯片的特异性为93.3％。结论：建立的结核分枝杆菌可视化抗体检测蛋白芯片检测敏感性显著高于痰涂片和痰培养方法,可用于结核病的临床辅助诊断,提高痰涂片和痰培养假阴性的检出率。     

Coevolution of the Monogenoidea (Platyhelminthes) based on a revised hypothesis of parasite phylogeny

A revised hypothesis for the phylogeny of the Subclass Polyonchoinea (Monogenoidea) was contructed employing phylogenetic systematics. The Acanthocotylidae (formerly of the Order Capsalidea) is transferred to the Order Gyrodactylidea based on this analysis. The new phylogeny is used to determine coevolutionary relationships of the familial taxa of Monogenoidea with their hosts. The coevolutionary analysis suggests that the Monogenoidea apparently underwent sympatric speciation or dispersal while parasitic on ancestral Guathostomata, resulting in two primary clades: the Polyonchoinea and the Oligonchoinea + Polystomatoinea. The two parasite clades apparently cospeciated independently with divergence of the Chondrichthyes and Osteichthyes. In the Polyonchoinea, the clade associated with Chondrichthyes experienced primary extiaction within the Holocephala, but coevolved into the Loimoidae and Monocotylidae in the Galeomorphii and Squalea (Elasmobranchii), respectively. Within the Osteichthyes, polyonchoineans experienced primary extinction with the divergence of Sarcopterygii, Polypteriformes and Acipenseriformes. They demonstrate primary dispersal from the Neopterygii into the Squalea (as Amphibdellatinea), Actinistia (as Neodactylodiscinea) and Urodela (as Lagarocotylidea). Secondary dispersals of polyonchoineans occurred in the Gyrodactylidae to the Polypteriformes, Urodela and Anura; in the Acanthocotylidae to the Myxinoidea and Squalea; in the Capsalidae to the Acipenseriformes and Elasmobranchii; and in the Monocotylidae to the Helocephala. The Oligonchoinea and Polystomatoinea developed upon divergence of the Chondrichthyes and Osteichthyes. Oligonchoineans cospeciated within the Chondrichthyes, with the Chimaericolidea developing within the Helocephala and the ancestor of the Diclybothriidea + Mazocraeidea within the Elasmobranchii. Two cases of primary dispersal occurred within this clade: the Diclybothriidae to the Acipenseriformes and the ancestor of mazocracidean families to the Neopterygii (both Osteichthyes). Secondary dispersal within the Oligonchoinea includes host switching of the common ancestor of Callorhynchocotyle (Hexabothriidae) to the Holocephala. Polystomatoineans coevolved within the Osteichthyes, but experienced primary extinctions in the Actinopterygii, Actinistia, Dipnoi and Amniota. Coevolution of the Sphyranuridae and Polystomatidae occurred with divergence of the Urodela and Anura, respectively. Secondary dispersal of polystomatids to the Urodela, Dipnoi and Amniota is suggested. A preliminary phylogenetic analysis of the Polystomatoinea suggests that primary extinction with secondary dispersal of polystomatids to the Dipnoi may not be necessary to explain extant parasite distributions, since Concinnocotyla (Concinnocotylinae) appears to represent the sister taxon of the remaining Polystomatidae + Sphyranuridae.     

益生菌混合发酵条件及其对发酵胡萝卜汁色泽及风味的影响

目的 研究3株益生菌混合发酵胡萝卜汁的发酵条件对色泽、风味的影响及其贮藏特性等.方法 通过菌种配比、单因素试验及正交试验优化了胡萝卜汁的发酵条件,并利用分光测色计和气质联用仪分别研究了发酵前后胡萝卜汁的风味成分和色泽变化.结果 添加30％ (m/m)的新鲜胡萝卜汁(原液),接种3％的以1∶1∶1(v/v/v)混合的嗜热...