The Immunomodulatory Action of Sialostatin L on Dendritic Cells Reveals Its Potential to Interfere with Autoimmunity

Sialostatin L (SialoL) is a secreted cysteine protease inhibitor identified in the salivary glands of the Lyme disease vector Ixodes scapularis. In this study, we reveal the mechanisms of SialoL immunomodulatory actions on the vertebrate host. LPS-induced maturation of dendritic cells from C57BL/6 mice was significantly reduced in the presence of SialoL. Although OVA degradation was not affected by the presence of SialoL in dendritic cell cultures, cathepsin S activity was partially inhibited, leading to an accumulation of a 10-kDa invariant chain intermediate in these cells. As a consequence, in vitro Ag-specific CD4(+) T cell proliferation was inhibited in a time-dependent manner by SialoL, and further studies engaging cathepsin S(-/-) or cathepsin L(-/-) dendritic cells confirmed that the immunomodulatory actions of SialoL are mediated by inhibition of cathepsin S. Moreover, mice treated with SialoL displayed decreased early T cell expansion and recall response upon antigenic stimulation. Finally, SialoL administration during the immunization phase of experimental autoimmune encephalomyelitis in mice significantly prevented disease symptoms, which was associated with impaired IFN-gamma and IL-17 production and specific T cell proliferation. These results illuminate the dual mechanism by which a human disease vector protein modulates vertebrate host immunity and reveals its potential in prevention of an autoimmune disease. The Journal of Immunology, 2009, 182: 7422-7429.

Cellular and molecular characterization of an embryonic cell line (BME26) from the tick Rhipicephalus (Boophilus) microplus

The cellular and molecular characteristics of a cell line (BME26) derived from embryos of the cattle tick Rhipicephalus (Boophilus) microplus were studied. The cells contained glycogen inclusions, numerous mitochondria, and vesicles with heterogeneous electron densities dispersed throughout the cytoplasm. Vesicles contained lipids and sequestered palladium meso-porphyrin (Pd-mP) and rhodamine-hemoglobin, suggesting their involvement in the autophagic and endocytic pathways. The cells phagocytosed yeast and expressed genes encoding the antimicrobial peptides (microplusin and defensin). A cDNA library was made and 898 unique mRNA sequences were obtained. Among them, 556 sequences were not significantly similar to any sequence found in public databases. Annotation using Gene Ontology revealed transcripts related to several different functional classes. We identified transcripts involved in immune response such as ferritin, serine proteases, protease inhibitors,. antimicrobial peptides, heat shock protein, glutathione S-transferase, peroxidase, and NADPH oxidase. BME26 cells transfected with a plasmid carrying a red fluorescent protein reporter gene (DsRed2) transiently expressed DsRed2 for up to 5 weeks. We conclude that BME26 can be used to experimentally analyze diverse biological processes that occur in R. (B.) microplus such as the innate immune response to tick-borne pathogens. (C) 2008 Elsevier Ltd. All rights reserved.

Proteins in the saliva of the Ixodida (ticks): Pharmacological features and biological significance

The saliva of ticks (Suborder Ixodida) is critical to their survival as parasites. A tick bite should result in strong responses from the host defence systems (haemostatic, immune and inflammatory) but tick saliva appears to have evolved to counter these responses. We review current knowledge of tick saliva components, with emphasis on those molecules confirmed to be present in the secreted saliva but including some that have only been confirmed to be present in salivary glands. About 50 tick saliva proteins that are well described in the literature are discussed. These saliva components include enzymes, enzyme inhibitors, amine-binding proteins and cytokine homologues that act as anti-haemostatic, anti-inflammatory or immuno-modulatory agents. Sequence comparisons are illustrated. The importance of tick saliva and the significance of the findings to date are also discussed. (C) 2006 Elsevier Ltd. All rights reserved.

Geographic variation of the Anderson's Niviventer (Niviventer andersoni) (Thomas, 1911) (Rodentia: Muridae) of two new subspecies in China verified with cranial morphometric variables and pelage characteristics

A total of 66 specimens of Niviventer andersoni with intact skulls was investigated on pelage characteristics and cranial morphometric variables. The data were subjected to principal component analyses as well as to discriminant analyses, and measurement

Use of plastic tips in artificial feeding of Dermacentor (Anocentor) nitens females Neumann, 1897 (Acari: Ixodidae)

The establishment of laboratory colonies of ticks is often hampered by their lack of adaptation to alternative hosts. The aim of this study was to artificially feed partially engorged Dermacentor (Anocentor) nitens females through plastic tips, and to identify what are the optimal conditions of application of this technique to get as much as possible close to the natural conditions. The technique of artificial feeding through plastic tips allowed the engorgement of D. nitens ticks to a final weight within the normal range for the species. (C) 2014 Published by Elsevier GmbH.

Questing Dermacentor reticulatus harbouring Babesia canis DNA associated with outbreaks of canine babesiosis in the Swiss Midlands

In 2011 and 2012, outbreaks of clinical canine babesiosis were observed in 2 areas of the Swiss Midlands that had no history of this disease so far. In one area, cases of canine babesiosis occurred over 2 consecutive tick seasons. The outbreaks involved 29 dogs, 4 of which died. All dogs were infected with large Babesia sp. as diagnosed in Giemsa-stained blood smears and/or PCR. These were identified as B. canis (formerly known as B. canis canis) by subsequent partial sequencing of the 18S rRNA gene of Babesia sp. Interestingly, the sequence indicated either a genotype with heterogeneity in the ssrRNA gene copies or double infection with different B. canis isolates. None of the dogs had a recent travel history, but one had frequently travelled to Hungary and had suffered twice from clinical babesiosis 18 and 24 months prior to the outbreak in autumn 2011. Retrospective sequencing of a stored blood DNA sample of this dog revealed B. canis, with an identical sequence to the Babesia involved in the outbreaks. For the first time in Switzerland, the partial 18S rRNA gene of B. canis could be amplified from DNA isolated from 19 out of 23 adult Dermacentor reticulatus ticks flagged in the same area. The sequence was identical to that found in the dogs. Furthermore, one affected dog carried a female D. reticulatus tick harbouring B. canis DNA. Our findings illustrate that, under favourable biogeographic and climatic conditions, the life-cycle of B. canis can relatively rapidly establish itself in previously non-endemic areas. Canine babesiosis should therefore always be a differential diagnosis when dogs with typical clinical signs are presented, regardless of known endemic areas.

Linefish resources annual report for the year 2000. Part 1: fisheries assessment

This work reflects the activities of line and trap fishing in Southern Mozambique in 2000. The catch in line fishing has been estimated at 441 mt, according to the DNAP records. The same sources indicated that 1767 days were spent at sea and the estimated catch rate was 250 Kg per boaticlay. Most of the line fishing effort shifted away from Maputo and moved to Inhambane region. The monthly analysis of fishing vessels, stricter controls over catch and effort data submission, development of long-term research programme and the continuation of the on board sampling to improve the data collection are the recommendation for line fishing. The catch of trap vessels increases from 30mt in 1997 to 172 mt in 2000, during which the total number of traps increased from 25 to 300. During this time the number of fishing days has remained relatively constant, as well the soak time. These data sets are thus not compatible with each other, reflecting an increase in daily catch from 243 Kg to 791 kg. The species composition is mainly dominated by P. coeruleopunctatus, C. puniceus, C. nufar and E. andersoni.

Pan-biogeografia dos gêneros Didelphis, Philander, Metachirus, Chironectes e Lutreolina (Didelphimorphia: Didelphidae)

A ordem Didelphimorphia, de marsupiais americanos, apresenta 19 gêneros e 96 espécies, todos membros da família Didelphidae, que é dividida em duas subfamílias, Caluromyinae e Didelphinae. A subfamília Didelphinae contém (não apenas) as tribos Didelphini e Metachirini. A tribo Didelphini compreende 15 espécies de quatro gêneros: Chironectes (1 espécie), Lutreolina (1), Didelphis (6) e Philander (7) e a tribo Metachirini é monotípica, com apenas uma espécie do gênero Metachirus. Estes cinco gêneros encontram-se distribuídos amplamente pelas Américas, desde o sul do Canadá até a região central da Argentina. O objetivo deste estudo foi buscar identificar e explicar, através de análise pan-biogeográfica, os padrões de distribuição das espécies destes cinco gêneros. Para tal, foi feito um levantamento em banco de dados, coleções científicas e artigos científicos para a obtenção de dados sobre as localidades de registro de cada espécie. Estas foram então marcadas em mapas e a partir destes, as localidades de ocorrência foram conectadas com linhas de menor distância para formação dos traços individuais. Pela sobreposição dos traços individuais chegou-se aos traços generalizados e do encontro destes, aos nós biogeográficos. Os pontos de ocorrência foram também plotados em mapas de biomas para análise. Encontramos três traços generalizados e dois nós biogeográficos, um no centro da Bolívia na província biogeográfica de Puna e outro na Argentina, na província de Misiones. Quatro espécies não participaram de nenhum dos traços generalizados, provavelmente devido à sua distribuição mais restrita (Philander deltae, P. andersoni, P. olrogi e P. mcilhennyi). Chironectes minimus e Metachirus nudicaudatus tiveram seus traços coincidentes com dois traços generalizados, o que está de acordo com suas divisões de subespécies. Identificamos os diferentes padrões existentes para o norte da América do Sul (Venezuela) já apontado por diversos autores, porém apenas quando analisadas as subespécies em separado. Alguns limites para a distribuição das espécies puderam ser identificados, como por exemplo o istmo de Tehuantepec, no México, para Chironectes minimus e Metachirus nudicaudatus e o limite da região neotropical para P. opossum e D. marsupialis. O limite de distribuição sul de Philander opossum e P. frenatus é provavelmente o rio Paraguai, que deve servir de barreira para o contato entre as duas espécies. A descaracterização dos ambientes naturais pelo desmatamento vem alterando os padrões naturais de distribuição das espécies, com o registro de espécies de áreas abertas em biomas de mata. Lutreolina crassicaudata apresenta distribuição disjunta, ocupando duas áreas de vegetação aberta, uma no noroeste e outra no centro e sudeste da América do Sul, padrão provavelmente gerado pelos períodos de retração e expansão de áreas de savana do Mioceno superior ao Holoceno, levando à captura destes enclaves de vegetação aberta, com seu isolamento por áreas de floresta. Os nós e traços generalizados aqui identificados coincidiram com os encontrados por outros autores. Apesar da pan-biogeografia poder ser usada para propor áreas de proteção ambiental, nos locais em que encontramos os nós biogeográficos já existem unidades de conservação, não havendo assim necessidade de propor novas áreas no caso desses marsupiais. Ainda existe uma grande necessidade de um melhor conhecimento da distribuição e taxonomia das espécies estudadas, o que promoveria um melhor entendimento dos padrões biogeográficos existentes

中国大陆白腹鼠属的分支系统发育研究

　运用Hennig 的分支系统学原理和方法, 对分布于中国大陆的白腹鼠属7 个种的部分外部及头骨形态学特征 进行性状数据分析, 结合应用Paup 和Hennig 86 两种分支系统学软件对它们之间的系统发育关系进行研究。结果表 明: 安氏白腹鼠N. andersoni 和川西白腹鼠N. excelsior 亲缘关系较近, 二者聚在一起, 组成了Musser (1981) 的安 氏白腹鼠种组N. andersoni2Division ; 另外刺毛鼠N. f ulvescens 和褐尾鼠N. cremoriventer 的亲缘关系最近, 二者先 聚在一起, 再与社鼠N. conf ucianus 聚在一起, 形成了安氏白腹鼠种组的姐妹群; 而梵鼠N. brahma 是上述5 种白 腹鼠组成类群的姐妹群, 灰腹鼠N. eha 则与前6 种白腹鼠组成类群互为姐妹群。

An analysis of the 1970 commercial fish catch in three areas of the Kafue Floodplain

The commercial landings of fish in three areas of the Kafue Floodplain were examined in regard to thhing technique used, catch per unit effort, and species composition and length size. Gillnets were used throughout the year although predominantly in the wet season, and drawnets (similar to beach seines) were used at periods of low water level. Fishermen used a varying number of gillnets in each area, and the catehes also varied according to month. Principal species caught on the floodplain were clarias gariepinus and Tilapia andersoni. There are indications that, whereas the catch per gillnet in the year's 1965-1970 may be lower than in the 1950s. The drawnet calch per unit of effort of these later years is higher than in the 1950s.

镇海棘螈繁殖群体生态和遗传结构研究

genetics, such as: population size, reproduction, mating system, growth, development,genetic structure and systematics status; The main results are presented below: The seasonal variation of the operational sexual ratio of this animal was found in the field and the ration always bias the female in the breeding season. Aiming at this character and considering the distance of time and space of both sexual habitat in breeding season, we census female population first by toe-clipping mark-recapture method, then estimated the population size with the definitive sexual ratio. Up to now, this species was found only at the Beilun district of the Ningbo City. The population size of the Ruiyan Temple Forest Park approximates to 369. The status of this population is extremely endangered, so besides protecting this population at the original locality, we also suggested to breed the salamander in fenced locality and to hatch embryos artificially, and send metamorphosed juveniles back to nature. We can transfer some individuals to other similar habitats or breed them under artificial conditions for saving this species from extinction. The early developmental stage of the Chinhai salamander is the same as its relative species, E. andersoni. Their balanceres are poorly developed and disappear very early. Temperature and moisture significantly influence the embryonic development of the Chinhai salamander. The embryonic stage is approx. 29 days under room temperature. The hatchling grows in a logarithmic curve. The larvae stage in water is approx. 58- 88 days. Many factors influence the nomal development, including two aspects of internal and external. Due to these factors, the effective protected measures were presented in detail. The breeding migration of E. chinhaiensis takes place at late March～late April every year. This salamander's hatching rate is high, but the rate of hatchling migrating into water is low. The average effectiveness of all the nest sites is 36.7%. The maternal self-conservation was contrary to the reproductive success of the egg-laying strategy. In the strategy of egg-laying behavior, the first factor selected by the female was its self-conservation, the second is embryonic survival rate, and the last is rate of hatchling survival rate. The oviposition selection is significant for the survival of the larvae. Based on the analysis of the evolutionary process of reproductive behaviors nad egg-laying site selections of all genera of the family Salamandridae, we deduced that perhaps Echinotriton is a transitional type in the evolutionary process from water to land. Due to its location in the adaptive stage in the terrestrial evolution, Echinotriton chinhaiensis's terrestrial nest may be one of important reason that causes this species to be endangered. The genetic deversity analysis shows that although the population size of the Chinhai salamander is quite small compared to other Chinese salamandrid species, the genetic diversity of this population is not reduce remarkably. We explain this phenomena with the polygamy mating system of this species. The result of 4 families' parenthood determinations shows that the parenhood determination can be taken without any paternal information. The "children" of every female include rich genetic information from at least two "fathers". It implies that female Chinhai salamander mates more than once with different males in a breeding season. The molecular evidence, the behavioral observation evidences and the sperm evidence in the female cloaca proved that this species has a polygamy mating system. The kin recognition in the mating of adult salamander was first discussed. The taxonomic status and phylogenetic relationships of 12 species representing 6 genera in the family Salamandridae were studied using DNA fingerprinting. The results showed that the DNA fingerprinting. The results showed that the DNA fingerprinting patterns demonstrated rich genetic diversity and species diversity, and also revealed the taxonomic status and phylogenetic relationshipes of higher taxa to a certain extent. The results are highly consistent with those obtained from the studies based on the morphology, ecology, cytology and molecular biology. The compreshensive analysis indicate that Tylototrition hainanensis and T. wenxianensis should be valid species; Echinotriton should be a valid genus;Tylotortriton is a natural cluster; Tylotortriton asperrimus should be put in Tylototrition rather than in Echinotriton, Hypselotriton and Allomestriton are synonyms of Cynops and Paramesotriton, respectively. There are three main groups in Chinese salamandride: Cynops, Paramesotriton and Pachytrition from the first group, the species of the Tylototriton from the second, and E. chinhaiensis composes the third.

TICK INNATE IMMUNITY

Ticks are blood feeding parasites transmitting a wide variety of pathogens to their vertebrate hosts. The vector competence of ticks is tightly linked with their immune system. Despite its importance, our knowledge of tick innate immunity is still inadequate and the limited number of sufficiently characterized immune molecules and cellular reactions are dispersed across numerous tick species. The phagocytosis of microbes by tick hemocytes seems to be coupled with a primitive complement-like system, which possibly involves self/nonself recognition by fibrinogen-related lectins and the action of thioester-containing proteins. Ticks do not seem to possess a pro-phenoloxidase system leading to melanization and also coagulation of tick hemolymph has not been experimentally proven. They are capable of defending themselves against microbial infection with a variety of antimicrobial peptides comprising lysozymes, defensins and molecules not found in other invertebrates. Virtually nothing is known about the signaling cascades involved in the regulation of tick antimicrobial immune responses. Midgut immunity is apparently the decisive factor of tick vector competence. The gut content is a hostile environment for ingested microbes, which is mainly due to the antimicrobial activity of hemoglobin fragments generated by the digestion of the host blood as well as other antimicrobial peptides. Reactive oxygen species possibly also play an important role in the tick-pathogen interaction. The recent release of the Ixodes scapularis genome and the feasibility of RNA interference in ticks promise imminent and substantial progress in tick innate immunity research.

Antimicrobial activity in the tick Rhipicephalus (Boophilus) microplus eggs: Cellular localization and temporal expression of microplusin during oogenesis and embryogenesis

Arthropods display different mechanisms to protect themselves against infections, among which antimicrobial peptides (AMPs) play an important role, acting directly against invader pathogens. We have detected several factors with inhibitory activity against Candida albicans and Micrococcus luteus on the surface and in homogenate of eggs of the tick Rhipicephalus (Boophilus) microplus. One of the anti-M. luteus factors of the egg homogenate was isolated to homogeneity. Analysis by electrospray mass spectrometry (ESI-MS) revealed that it corresponds to microplusin, an AMP previously isolated from the cell-free hemolymph of X (B.) microplus. Reverse transcription (RT) quantitative polymerase chain reactions (qPCR) showed that the levels of microplusin mRNA gradually increase along ovary development, reaching an impressive highest value three days after the adult females have dropped from the calf and start oviposition. Interestingly, the level of microplusin mRNA is very low in recently laid eggs. An enhance of microplusin gene expression in eggs is observed only nine days after the onset of oviposition, achieving the highest level just before the larva hatching, when the level of expression decreases once again. Fluorescence microscopy analysis using an anti-microplusin serum revealed that microplusin is present among yolk granules of oocytes as well as in the connecting tube of ovaries. These results, together to our previous data. suggest that microplusin may be involved not only in protection of adult female hemocele, but also in protection of the female reproductive tract and embryos, what points this AMP as a considerable target for development of new methods to control R. (B.) microplus as well as the vector-borne pathogens. (c) 2009 Elsevier Ltd. All rights reserved.

Factors associated to Theileria equi in equids of two microregions from Rio de Janeiro, Brazil

Amostras de soro de 714 equídeos das microrregiões de Itaguaí e Serrana, Rio de Janeiro, Brasil, foram submetidas ao teste de imunofluorescência indireta (título 1:80) para Theileria equi. A prevalência entre as microrregiões e os fatores associados à soropositividade foram avaliados e a razão de prevalência (RP) calculada. A prevalência geral para T. equi foi de 81,09% (n = 579), com maior prevalência (p < 0,05) para microrregião de Itaguaí (85,43%), quando comparado a Serrana (76,92%). A região, altitude, nível da propriedade e origem dos equídeos foram associados (p < 0,05) com a soropositividade para T. equi. Equídeos criados na microrregião de Itaguaí (RP = 1,11; p = 0,003) e em altitudes abaixo de 500 m (RP = 1.10; p = 0,014) apresentaram maior chance de se tornarem soropositivos para T. equi. Além disso, quando são criados em propriedades de nível ruim (RP = 1,13; p = 0,018) e nascidos na propriedade (RP = 1,10; p = 0,008) apresentaram mais chance de terem contato com T. equi. As principais espécies de carrapatos encontradas parasitando os equídeos foram Amblyomma cajennense e Dermacentor (Anocentor) nitens. As microrregiões estudadas são endêmicas para theileriose equina e de estabilidade enzoótica para T. equi. Apenas os fatores relacionados à área de coleta das amostras de soro influenciaram a soropositividade dos equídeos para T. equi naquela região.

Detection of Cryptosporidium parvum oocysts in calf fecal samples by direct immunofluorescence assay

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)