Accumulation of protoporphyrin-IX (PpIX) in leukemic cell lines following induction by 5-aminolevulinic acid (ALA)

We investigated the amounts of protoporphyrin IX (PpIX) accumulated in noninduced cells and following 5-aminolevulinic acid (ALA)-induction. Following ALA administration PpIX increased in all leukemic cell lines under investigation (HEL 26-fold, HL60 6-fold, Jurkat 3-fold, ML2 2-fold) but not in lymphocytes. Compared to other cell lines studied, HEL cells showed the lowest basal level of PpIX and the largest relative increase in PpIX. Despite a high increase following ALA treatment, the PpIX level in HEL cells is almost as low as in lymphocytes. It is in agreement with their relatively low sensitivities of ALA-induced photodynamic therapy (ALA-PDT) shown previously [(Grebenová, D., Cajthamlová, H., Bartosová, J., Marinov, J., Klamová, H., Fuchs, O., Hrkal, Z., 1998. Selective destruction of leukemic cells by photo-activation of 5-aminolevulinic acid induced protoporphyrin IX. J. Photochem. Photobiol. B: Biol. 47, 74-81)]. The ferrochelatase activities in the individual cell lines are in good inverse correlation with PpIX amounts accumulated in the ALA-induced cells, but not with the relative increase (ratio) of PpIX levels from basal to ALA-induced ones. This is most apparent in HEL cells and lymphocytes. There is probably different regulation of heme biosynthesis in erythroid cells, which are therefore not suitable for the studies of ALA-PDT mechanism. PpIX was accumulated more extensively in absence of fetal calf serum than in its presence. The amounts of PpIX accumulated in cells decreased exponentially with increasing fetal calf serum concentration.     

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.     

Intraspecific variability of Bipolaris sorokiniana isolates determined by random-amplified polymorphic DNA (RAPD)

Isolates of Bipolaris sorokiniana were analyzed by random-amplified polymorphic DNA (RAPD) techniques to determine the amount of intraspecific genetic variability and to study host-pathogen interactions. Ten isolates originated from different regions of Brazil were examined. Plants of the wheat cultivars BR8, BH1146 (original host) and IAC-5 Maringá, classified as resistant, moderately resistant or susceptible to B. sorokiniana, respectively, were inoculated with these 10 isolates. Twenty-seven isolates were recovered from these cultivars and were analyzed by RAPD assay and compared to the RAPD of the original 10 isolates. According to the RAPD profiles there was a high level of genetic variability among the isolates. We detected 69 polymorphic fragments, ranging from 1.6 to 0.54 kb, in the original 10 isolates; 57 fragments with sizes between 1.98 and 0.38 kb from the isolates recovered from BH1146; 47 polymorphic bands, ranging from 1.96-0.54 kb, were detected in the isolates from BR8 and 32 fragments between 1.98 and 0.42 kb in isolates were recovered from IAC-5 Maringá. The number of polymorphic fragments varied, even for the same isolate, when the isolates were recovered from different cultivar hosts.     

Molecular differentiation of species of the genus Zungaro (Siluriformes, Pimelodidae) from the Amazon and Paraná-Paraguay River basins in Brazil

Fish species of the Zungaro genus (Siluriformes, Pimelodidae) are amongst the largest migratory fish in Latin America and have considerable economic importance for commercial fishing in Brazil. However, natural populations of this large catfish are experiencing a severe decline. There are significant taxonomical inconsistencies for this fish. Two geographically separated species of the fish were initially described, one endemic in the Amazon and another in the Paraná-Paraguay River basins. A taxonomic review had recently proposed that there is only one Zungaro species in Brazil, based on morphological data. We made a molecular study of Zungaro populations in an attempt to solve taxonomical inconsistencies and to analyze genetic diversity in natural populations of this genus. We analyzed two regions of the mitochondrial DNA (the control region and the ATPase 6 gene region) of individuals sampled from the Paraná-Paraguay River and Amazon River basins. Analyses based on p-distances and maximum likelihood phylogenetic models showed a genetic difference between populations corresponding to different species. Genetic differentiation between Zungaro populations was at the same level as that observed between other Siluriformes species, using the same DNA sequences. We conclude that Zungaro species of the Paraná-Paraguay River basin do not belong to the same species found in the Amazon basin. This finding has a significant implication for conservation of this fish, given that populations are disappearing at a high rate in the Paraná-Paraguay River basin, mainly due to impoundments.     

New species of Tetramorium Mayr (Hymenoptera: Formicidae) from Puebla State, Mexico

Tetramorium notomelanum sp. n. is described from the Tehuacán Valley, state of Puebla, México. Its distribution and relation with other species of the tortuosum-group is discussed. The new species of Tetramonium is described from workers, and distinguished from others of the group by several characters: i) black coloration of the body; ii) size: T. notomelanum sp. n. is smaller than T. hispidum (Wheeler), T. mexicanum Bolton and T.spinosum (Pergande), but larger than T. bicolorum Vásquez-Bola?os and T. placidum Bolton; iii) length of the hairs of the dorsal of the head are equal to the diameter of eye; iv) the length of the hairs on the scape and tibiae less than the width of the appendage where they are located. This is the second species of the tortuosum group of Tetramorium found in the State of Puebla, and the fourth recorded in Mexico.     

Molecules, wing pattern and distribution: an approach to species delimitation in the "loxurina group" (Lepidoptera: Lycaenidae: Penaincisalia)

The wide range of morphological variations in the "loxurina group" makes taxa identification difficult, and despite several reviews, serious taxonomical confusion remains. We make use of DNA data in conjunction with morphological appearance and available information on species distribution to delimit the boundaries of the "loxurina" group species previously established based on morphology. A fragment of 635 base pairs within the mtDNA gene cytochrome oxidase I (COI) was analysed for seven species of the "loxurina group". Phylogenetic relationships among the included taxa were inferred using maximum parsimony and maximum likelihood methods. Penaincisalia sigsiga (Bálint et al), P. cillutincarae (Draudt), P. atymna (Hewitson) and P. loxurina (C. Felder & R. Felder) were easily delimited as the morphological, geographic and molecular data were congruent. Penaincisalia ludovica (Bálint & Wojtusiak) and P. loxurina astillero (Johnson) represent the same entity and constitute a sub-species of P. loxurina. However, incongruence among morphological, genetic, and geographic data is shown in P. chachapoya (Bálint & Wojtusiak) and P. tegulina (Bálint et al). Our results highlight that an integrative approach is needed to clarify the taxonomy of these neotropical taxa, but more genetic and geographical studies are still required.     

Nyssomyia intermedia (Lutz & Neiva, 1912) and Nyssomyia neivai (Pinto, 1926) (Diptera: Psychodidae: Phlebotominae) geographical distribution and epidemiological importance

Nyssomyia intermedia (Lutz & Neiva 1912) and N. neivai (Pinto 1926) are possible vectors of tegumentary leishmaniasis in some regions of Brazil. Further, the latter was until recently, considered a junior synonym of the former. This study has the purpose of updating our knowledge of the geographical distribution of these species, based on specimens deposited at the collection of the Centro de Pesquisas René Rachou-Fiocruz, Faculdade de Saúde Pública-Universidade de S?o Paulo, and on data presented by literature as also to associate this distribution with the cutaneous leishmaniasis cases reported. It has been reported that N. intermedia occurs in the states of the Northeastern Region, in Rio de Janeiro, Espírito Santo, on the northern coast of S?o Paulo, in eastern Minas Gerais, Mato Grosso do Sul, and Goiás, close to the border with Minas Gerais and Bahia. N. neivai occurs in the Southern Region, southern coast and in western S?o Paulo, southern and western Minas Gerais, southern Goiás, and southern Pará, beyond Argentina, Bolivia, and Paraguay. It is important to highlight that N. intermedia and N. neivai occur in sympatry in Minas Gerais and S?o Paulo. N. intermedia or N. neivai are predominant or are captured abundantly in several cutaneous leishmaniasis foci in the Southeastern and Southern regions of Brazil.     

Population genetics analysis of Podocnemis sextuberculata (Testudines, Podocnemidae): lack of population structure in the central Amazon Basin

The chelonians are, in general, important for the economy of the traditional populations of the Amazon region, especially as a source of animal protein. Furthermore, sub-products, such as eggs and fat, are utilized in the manufacture of cosmetics, and the plastron and carapace are used in the manufacture of adornments. The freshwater turtle species Podocnemis sextuberculata, locally known as "ia?á" or "pitiú", is widely distributed in the Amazon Basin in Brazil and also in Colombia and Peru. This species is on the International Union for Conservation of Nature Red List in the category of vulnerable species. We examined the genetic variability and population structure of three populations represented by 64 individuals sampled from Reserva Federal de Abufari, Tapauá, Amazonas State; Reserva de Desenvolvimento Sustentável Mamirauá, Tefé, Amazonas State, and Terra Santa, Pará State. All of these are over 1000 km from each other. A partial 415-bp sequence of the mitochondrial gene ND1 was utilized as a molecular marker. Seven haplotypes were observed; the most common haplotype was shared by all the areas sampled, while the rarest haplotypes were represented by a single individual and were thus restricted to a single locality. The sharing of the most common haplotype, the high number of migrants (Nm) and the AMOVA results indicate a lack of genetic structure among the sampling localities. The levels of genetic variability observed were homogeneous among the sampling localities. These results (?(ST) and Nm) are compatible with what is known about the ecology of this species, which has a great migratory capacity.     

Historical sketch of Slovak Haban (Hutterite) population based on autosomal STR analysis

According to the Hutterite chronicles, the Habans arrived from Austrian Tyrol, Switzerland, and northernmost Italy and stayed in four regions of Slovakia (Soboti?te, Vel'ké Leváre, Moravsky Sv?ty Ján, Tren?ín). There are some communities in western Slovakia that retained their Haban cultural identity and still identify themselves as descendents of the Hutterite population with their own specific customs. Slovak Habans are typical founder population with significant social isolation for which high degree of inbreeding is typical. Present study investigated STR polymorphisms as a powerful genetic tool for population genetic studies. The aim was to perform a comparative, population genetic study based on 15 STR loci widely used in forensic genetics, of the Haban population, the Slovak majority population and the population of Tyrol. We analyzed allele frequencies and other statistical parameters in three selected populations in order to identify groups of specific ethnic origin and establish their genetic relationship. The data set included 110 unrelated Habans and 201 unrelated individuals from the Slovak majority population, as well as allelic frequencies for the population of Austrian Tyrol available in the literature. Population pairwise FST values used as a short term genetic distance between populations showed significant differentiation between the Habans and both reference populations (FST=0.0025 and 0.0042 for comparison with the Slovaks and Austrians, respectively; p<10(-3)). The Slovak Hutterites were demonstrated to be genetically distinct and more closely related to their geographic neighbors than to their historical ancestral population, which may be at least partially explained by gene flow between neighboring Haban and Slovak populations.     

Carcinops troglodytes (Erichson) (Coleoptera: Histeridae) preying on Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) in poultry houses

We first record Carcinops troglodytes (Paykull) as a predator of Alphitobius diaperinus (Panzer) larvae in poultry houses in North and Northwest regions of the state of Paraná. Carcinops spp. are commonly recorded as predators of dipterans, and this record in poultry houses indicates the possibility of exploiting such predator for future studies aiming the development of management strategies for A. diaperinus.     

Genetic variability among populations of Lutzomyia (Psathyromyia) shannoni (Dyar 1929) (Diptera: Psychodidae: Phlebotominae) in Colombia

Polyacrylamide gel electrophoresis was used to elucidate genetic variation at 13 isozyme loci among forest populations of Lutzomyia shannoni from three widely separated locations in Colombia: Palambí (Nari?o Department), Cimitarra (Santander Department) and Chinácota (Norte de Santander Department). These samples were compared with a laboratory colony originating from the Magdalena Valley in Central Colombia. The mean heterozygosity ranged from 16 to 22%, with 2.1 to 2.6 alleles detected per locus. Nei's genetic distances among populations were low, ranging from 0.011 to 0.049. The estimated number of migrants (Nm=3.8) based on Wright's F-Statistic, F ST, indicated low levels of gene flow among Lu. shannoni forest populations. This low level of migration indicates that the spread of stomatitis virus occurs via infected host, not by infected insect. In the colony sample of 79 individuals, the Gpi locus was homozygotic (0.62/0.62) in all females and heterozygotic (0.62/0.72) in all males. Although this phenomenon is probably a consequence of colonization, it indicates that Gpi is linked to a sex determining locus.     

Characterization of 10 microsatellite loci in an avian louse, Degeeriella regalis (Phthiraptera: Ischnocera: Philopteridae)

We isolated and characterized 10 polymorphic microsatellite loci in an ischnoceran louse, Degeeriella regalis, which parasitizes the threatened Galápagos hawk (Buteo galapagoensis) and other falconiform birds. The loci were screened across 30 individuals from two island populations in the Galápagos Islands. The number of alleles per locus ranged from two to 28. Polymorphic information content ranged from 0.14 to 0.94 and observed heterozygosity ranged from 0 to 0.67. These markers will be valuable in comparative population genetics studies in this species, which is the focus of a long-term population and disease ecology research program.     

Morphology of Philometroides barbi (Nematoda: Philometridae), a rare tissue parasite of the Mediterranean barbel Barbus meridionalis (Osteichthyes)

A female specimen of the nematode Philometroides barbi Moravec, Simková, Hanzelová, Spakulová et Caki?, 2005, a little-known histozoic parasite of the Mediterranean Barbus meridionalis, was recorded from the fin of its fish host in Bulgaria. Scanning electron microscopical examination, used for the first time in this species, made it possible to determine the character of cephalic papillae (14 papillae arranged in 2 circles) and amphids and confirmed the presence of 4 large caudal lobes. These features distinctly distinguish P. barbi from Philometroides cyprini and other congeners parasitizing European fishes.     

Origin and population history of a recent colonizer, the yellow warbler in Galápagos and Cocos Islands

The faunas associated with oceanic islands provide exceptional examples with which to examine the dispersal abilities of different taxa and test the relative contribution of selective and neutral processes in evolution. We examine the patterns of recent differentiation and the relative roles of gene flow and selection in genetic and morphological variation in the yellow warbler (Dendroica petechia aureola) from the Galápagos and Cocos Islands. Our analyses suggest aureola diverged from Central American lineages colonizing the Galápagos and Cocos Islands recently, likely less than 300 000 years ago. Within the Galápagos, patterns of genetic variation in microsatellite and mitochondrial markers suggest early stages of diversification. No intra-island patterns of morphological variation were found, even across steep ecological gradients, suggesting that either (i) high levels of gene flow may be homogenizing the effects of selection, (ii) populations may not have had enough time to accumulate the differences in morphological traits, or (iii) yellow warblers show lower levels of 'evolvability' than some other Galápagos species. By examining genetic data and morphological variation, our results provide new insight into the microevolutionary processes driving the patterns of variation.     

Raphidascaris (Sprentascaris) lanfrediae sp. nov. (Nematoda: Anisakidae) from the fish Satanoperca jurupari (Osteichthyes: Cichlidae)

Raphidascaris (Sprentascaris) lanfrediae sp. nov. is described from the intestine of the freshwater fish Satanoperca jurupari (Heckel) (Cichlidae) from the Guamá River, state of Pará, Brazil. The prevalence in fish (n = 59) was 27% with intensity of one-124 (mean 16) nematodes per fish. The new species is characterized mainly by the markedly larger size of ventricular appendix in relation to the oesophagus, presence of short male caudal alae, 14-16 subventral pairs of preanal papillae and six pairs of postanal papillae.     

Paralytic shellfish poisoning (PSP) in Margarita Island, Venezuela

A severe outbreak of Paralytic Shellfish Poisoning (PSP) occurred in Manzanillo and Guayacán, northwestern coast of Margarita Island, Venezuela, between August and October 1991. A bloom of dinoflagellates including Prorocentrum gracile, Gymnodinium catenatum and Alexandrium tamarense seemed to be responsible for this outbreak. Levels of PSP toxins in mussels (Perna perna) exceeded the international safety limit of saxitoxin, 80 microg STX/100 microg meat. PSP toxin values varied between 2548 and 115 microg STX/100 g meat in Manzanillo, and between 1422 and 86 microg STX/100 g meat in Guayacán. At both locations, the highest levels were detected in August, when 24 patients exhibited typical symptoms of PSP toxicity after consuming cooked mussels (16 required hospitalization). A high pressure liquid chromatographic (HPLC) procedure was recently used on the 1991 samples. The major toxin detected in samples of both locations was decarbamoyl saxitoxin (dcSTX), but low concentrations of saxitoxin were also found in Manzanillo samples. Gonyautoxins GTX1, GTX2 and GTX3 were detected only at Guayacán, while in both locations, decarbamoylgonyatouxin (dcGTX2,3) toxins were detected. These findings represent the first time that causative toxins of PSP in Venezuela have been chemically identified, and confirm the presence of dcSTX and dcGTX in mussels from the Caribbean Sea. The presence of dcSTX and dcGTX in shellfish is indicative that Gymnodinium catenatum was a causative organism for outbreak of PSP.     

Genetic diversity in two populations of the snail Strombus gigas (Gastropoda: Strombidae) from Yucatan, Mexico, using microsatellite

The pink conch Strombus gigas is an important fisheries resource in the Caribbean region, including the Yucatán Peninsula. We analyzed the genetic diversity and genetic structure of two populations (Alacranes Reef and Chinchorro Bank) with the use of five microsatellite molecular markers. The results indicate that the two populations are in the same rank of genetic diversity (He), from 0.613 to 0.692. Significant deviation from H-WE was observed in the both populations due to deficit to heterozygotes, this was attributed to inbreeding as a consequence of over-fishing; nevertheless, other possible causes considered are mixing of individuals from two or more populations, and the existence of null alleles. Levels of genetic differentiation indicated the existence of a single homogenous population in the Yucatan Peninsula (F(ST) de 0.003, p = 0.49), which fits with highest levels of gene flow is significant (2.3 individuals) between both populations. Results from this study support the hypothesis that S. gigas is part of a single panmictic population in the Yucatan Peninsula; therefore, this fishery resource should be regulated the same way for both areas.     

中国巴蝶蛉属分类研究(脉翅目,蝶蛉科)

对中国巴蝶蛉属Balmes Navás,1910进行了分类研究和厘定,确认巴蝶蛉属中国有5种:滇缅蝶蛉Balmes birmanus Maclachlan,集昆蝶蛉Balmes chikuni sp.nov.,丽东蝶蛉Balmes formosus(Kuwayama),显赫蝶蛉Balmes notabilis Navás和川贵蝶蛉Balmes terissinus Navás.提供了这5个种的修订名录及分种检索表,对新种进行了描述,附形态特征影像.     

RAPD and mitochondrial DNA analysis of the soybean stalk weevil, Sternechus subsignatus (Coleoptera: Curculionidae)

Sternechus subsignatus Boheman (Curculionidae: Sternechini) is one of the primary Curculionidae species that reduces soybean yield in Brazil. Initially, outbreaks were reported in southern Brazil in 1973; but, more recent, outbreaks were reported in Bahia (summer 1997-1998) and Maranh?o (summer 2003-2004), two states in northeastern Brazil. A putative related species, S. pinguis (Fabricius), was first detected in Salta Province, Argentina. The objective of this study was to evaluate intraspecific molecular polymorphisms of geographically distinct Sternechus populations. Randomly amplified polymorphic DNA (RAPD) profiles and partial mitochondrial cytochrome B (CytB) gene sequences were used to determine whether individual soybean stalk weevils were one of two different species and to infer pest invasion pattern. Putative S. pinguis and S. subsignatus populations were collected in San Agustin (Cruz Alta, Tucumán Province, Argentina) and different sampling sites in the Brazilian states of Paraná, Bahia and Maranh?o. Polymorphic bands were obtained by RAPD and analyzed by Dice coefficients. Populations from southern Brazil were more closely related genetically to an Argentinean group than the populations sampled in northeastern Brazil. The Londrina Co., Brazil population displayed the highest intra-population genetic similarity. Most of the soybean stalk weevils collected from San Agustin, Tucumán, Argentina were divergent from those collected in Brazil. Sequencing and parsimony analysis of CytB did not differentiate specimens collected in Argentina and Brazil. Thus, our data show that soybean stalk weevil outbreaks and population increases in northeastern Brazil involved local genotypes.     

Simulium (Chirostilbia) brunnescens (Diptera: Simuliidae) - new species from the Brazilian cerrado, Manso Dam, state of Mato Grosso, Brazil

A new species of Simuliidae, Simulium (Chirostilbia) brunnescens, was discovered at Chapada dos Guimar?es, state of Mato Grosso, Brazil, and nearby municipalities (Paranatinga, Rosário do Oeste and Nobres). This species is described here based on the adults, pupae and larvae. This species is closely related to Simulium (C.) subpallidum Lutz, but could be differentiated in all stages: females, leg colour pattern and frontal dilatation size; males, gonostyle shape; pupae, number of gill filaments; larvae, body size and colour, postgenal cleft, ratio between antenna and stalk of labral fan.