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This paper reviewed the main mosquito-species resource of China recorded during 1828-2002 in different periods through Feng(1938),Meng(1955),Lu(1977),to Qu(2002).According to the new systematic series of Reinert(2001),the mosquito-record of China would be totally: 21 genera,52 subgenera,and 395 species/ subspecies up to 2006.The first report of mosquito studies in China including: new species described by foreigner or Chinese entomo-logists,check list,hand-book of "key to Chinese mosquitoes",and "Fauna of China" were cited.And with some discu-ssions on guarantee of the quality of resource materials,its utilization and resource-sharing.
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Objective To isolate and identify genes related to malaria parasite infection in vector mosquito, and to explore the mechanisms. Methods Anopheles stephensi infected with Plasmodium yoelii was used as tester (T) group, while uninfected but normal blood fed as driver (D) one. Engorged female mosquitoes of two groups were collected separately at 24 hours after biting. An enriched subtractive cDNA pool was generated through the course of suppression subtractive hybridization (SSH) and selective PCR amplification. The subtracted library was screened by hybridization using T and D cDNA mixture as probes, respectively. The positive clones, which produced stronger signal when probed with T than with D, were sequenced and their sequence homologues in GenBank database were searched with BLAST by internet. Results The analysis of subtraction efficiency showed that the differentially expressed genes in T comparing to in D were enriched significantly. In dot blot screening, 24 of 58 randomly selected clones (41.4%) were shown up regulation in malaria infected mosquitoes. The BLAST search of 23 genes revealed that 12 were homologous to functionally known genes, 4 were homologous to functionally unknown entries, and 7 were novel without any relatives. Nine of the 23 genes (39.1%) also hit homologous sequences in the An. gambiae EST database generated from an immune competent cell line treated with lipopolysaccharide (LPS). Conclusion An enriched cDNA pool of the mosquito genes which up regulated responsively at the early stage of malaria parasite infection was obtained. Expression screening against the pool indicated that various biochemical processes and mechanisms might be involved in the response of mosquito to parasite infection, especially those related with the innate immune system and energy metabolism.
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Objective To establish the molecular identification of five members in Anopheles maculatus complex from China. Methods Different rDNA-ITS2 regions of An. maculatus complex were sequenced and analyzed. The species specific primers were designed, and PCR assay was used for the identification. Results The length and GC contents of ITS2 were 328 bp, 58.54% in An. pseudowillmori, 330 bp, 57.85% in An. maculatus, 337 bp, 59.05% in An. willmori, 334 bp, 58.68% in An. dravidicus, and 338 bp, 57.69% in An. sawadwongporni, respectively. The intra-species ITS2 sequences were conservative. The ranges of divergence level among five members were from 9.7% to 18.9% . Five distinct specific fragments were amplified by PCR assay using five species specific primers and 5. 8S primer. The length was 119, 186, 231, 327 and 406 bp respectively. Conclusion The diagnostic PCR assay based on ITS2 divergence to distinguish five members of An. maculatus complex was simple and reliable.
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Objective] To clarify the taxonomic status of Anopheles lesteri and An.anthropophagus in China. [Methods] Using molecular identification (PCR assay and rDNA\|ITS2 sequencing) to examine the field anopheline mosquito specimens from Liaoning and Shandong. According to the ITS2 sequences, molecular phylogenetic tree was made. [Results] According to the molecular identification, An.lesteri and An.anthropophagus were distributed both in Liaoning Province and Shandong Province. The length and GC content of rDNA\|ITS2 sequence were 451 bp, 46 2% in An.lesteri (n=6), and 448 bp, 46 0% in An.anthropophagus (n=10), respectively. The ITS2 sequences from presentation sites were same in An.lesteri, while the intraspecies difference in An.anthropophagus was 0 88%. The specific difference between An.lesteri and An.anthropophagus was 25 7%. By analyzing molecular phylogenetic tree, the relationship between An.lesteri and An.sinensis, An.anthropophagus and An.liangshanensis was found to be closer. [Conclusion] According to the molecular identification, it was defined that An.lesteri and An.anthropophagus were sympatric independent species in China.
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AIM:To ascertain the species status of the Anopheles dirus from Hainan Province,Chi- na.METHODS:The nucleotide sequence of the second internal transcribed spacer(ITS2 ) of PCR- amplified r DNA was determined for the An.dirusspecimens which included 2 individu- als of the species A colony (AFRIMS) from Thailand and 5 individuals from Hainan Province.RESUL TS:A841 bp fragment was amplified from single mosquito.The fragment included the ITS2 and small portions of flanking 5 .8S (96 bp) and 2 8S (2 9bp) genes.The ITS2 was71 6 bp in length,the sequence was identical for all7individuals from both Hainan Province and Thailand.No evidence of intraspecies or intrapopulation variation was detected in the ITS2 and flanking regions.CONCL USION:The result suggests the existence of An. dirus A in Hainan Province,which was in agreement with previous studies from cytogenetic analysis and egg characteristics determined by scanning electron microscopy.
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This paper reports the rectification results of the tribe aedini mosquitoes formerly recorded in China, using the classification system proposed by Reinert during the recent years. Among all the 171 species of Chinese aedini mosquitoes examined, 160 species could be included in the new classification system. The other 11 species were listed in traditional taxonomic status for further study. The proposed new classification system of the Chinese aedini mosquitoes contained 29 genera, i.e. Aedes, Armigeres, Ayurakitia, Bothaella*, Bruceharrisonius*, Christophersiomyia*, Collessius*, Danielsia*, Downsiomyia*, Edwardsaedes*, Finlaya*, Fredwardsius*, Gilesius*, Heizmannia, Himalaius*, Hopkinsius*, Hulecoeteomyia*, Jihlienius*, Kenknightia*, Luius*, Mucidus*, Neomelaniconion*, Ochlerotatus, Phagomyia*, Scutomyia*, Stegomyia*, Tanakaius*, Udaya, and Verrallina. Among them, 22 genera (*) were new records in China. Besides, the authors made a significant revision to the following 4 species recorded formerly in 《Fauna Sinica, Insecta Vol. 8, Diptera: Culicidae》: Ae. (Edw.) antuensis as the synonym of Ed. pingpaensis, while Ae. (Sin.) occidentayunnanus, Ae. (Och.) flavidorsalis, and Ae. (Fin.) subsimilis should be rectified as Hz. (Mat.) occidentayunnana, Oc. albineus, and Ud. subsimilis, respectively.
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Objective To determine sequence of sporogony stage-specific (S type) 18S ribosomal RNA gene of Plasmodium yoelii (P.y) By265 strain, and by using it to detect the malaria parasites within vector mosquito. Methods A pair of conserved DNA primers, universe primer (Pu) and reverse transcription one (Pr), was designed and synthesized according to sequence of the 18S rRNA gene of Plasmodium berghei (P.b). The segment of the S type 18S rDNA of P.y was amplified by reverse transcript-polymerase chain reaction (RT-PCR) from dissected midguts of Anopheles stephensi infected with P.y on the 7th day after infective blood-meal, and its sequence was then determined. One P.y sporogony stage-specific primer (Pys) was selected according to the sequence. Using this primer and Pr, the parasites within mosquitoes were semi-quantitatively detected through RT-PCR between 1-7 d post-infection. Results The length of the amplified segment was 920 bp. Alignment in match region of the 18S rDNA among S type of P.y (PyS), S type of P.b (PbS) and asexual blood stage-specific one of P.y (PyA) revealed that the similarity between the former and the latter two reached 95^3% and 94^0% respectively. The density of amplified band was significantly concordance with the intensity of oocyst in the midgut. Sensitivity of RT-PCR method was higher than that of the traditional dissection and oocyst observation also. The assay could detect the 18S rRNA molecule of the parasites on the third day post-infection while their oocysts were difficult to be recognized under an optical microscope at that time. Conclusion This S type 18S rDNA sequence in P.y species was first reported (AF266261). As a molecular marker, it could be applied to monitoring the parasite development in its vector at an earlier stage semi-quantitatively with an adequate sensitivity and specificity.
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AIM:To diagnose a patient with clinical symptoms of fever,cough and asthma.ME- THODS:The sputum of the patient was subjected to microscopic examination,and the clini- cal pictures of the patientwas analysed.RESULTS:The red blood- like substance in the spu- tum was preliminarily defined as parasitic nematodes.Species identification indicated that they were a pair of Mammomonogamus laryngeusadult worms in copulation. The patient had obvious signs and symptoms of respiratory tract infection and eosinophilia of peripheral blood. Detection of the worms or eggs is the main base for diagnosis. CONCLUSION:It is the first record of human Mammomonogamus laryngeus infection in China.
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A 54-kDa protein overexpressed by chloroquine-resistant Plasmodium berghei ANKA strain was first reported by us. In this paper, the localization of this protein by immunoelec-tron microscopy is presented. The results showed that the protein was mainly scattered inside the cytoplasm of the early date trophozoites and schizonts of erythrocytic stage of P. berghei ANKA strain , and some of it was also found in cytoplasm of erythrocytes infected with parasites. The protein content was much higher in chloroquine-resistant P. berghei ANKA strain than in chloroquine-sensitive P. berghei ANKA strain, suggesting the importance of this protein in understanding mechanism of chloroquine resistance in malaria parasites .
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This paper presents a new revised "Checklist of the Anopheline Mosquitoes in China" based on the development of the mosquito-taxonomic researches during the years of 1988-2007. The new checklist contained 61 species (subspecies) of anopheline mosquitoes all in China. Twelve species among the past records were omitted because of their invalid specific names which were allocated into following categories: ① A doubtful record in China, with no typical specimen up to date since last century, e.g. Anopheles campestris reported in Yunnan; ② Misidentification: An. atroparvus and An. indiensis; ③ Confirmed as synonyms by hybridizing experiments or molecular identification, including 9 species as follows: An. changfus, An. dazhaius, An. kiangsuensis, An. anthropophagus, An. kunmingensis, An. xiaokuanus, An. junlianensis, An. yutsushiroensis (part) and An. fluviatilis. Meanwhile, the following rectified 4 anopheline mosquito species should be added to the new checklist: An. belenrae, An. lesteri, An. pullus, and An. baimaii.
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This paper deals with the taxonomic status and specific names of Anopheles anthropophagus and An. lesteri, the important malaria transmitting vector in China. Based on a historical review of the literature recorded from the country, substantial evidence from morphological and molecular biological studies gives reason to convince that An. anthropophagus is a synonym of An. lesteri. A resurrection of the specific name of An. lesteri Baisas et Hu, 1936 brooks no delay.
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The present review deals with the representative research papers on human parasites and parasitic diseases in China over the past hundred years (1871-2006). As the views focused on the development of the medical parasitology,the historical background and progressive characters in the period of fermentation,origination,and expansion have been discussed. The check list of the first cases of human parasitic diseases reported in China during 1871-2006 contained 128 species of parasitic pathogens,and among them 38 species were the newly revisional records. The citation from Faust's paper(1923) proved that previous record of "the first case of Eurytrema pancreaticum from Hongkong" was an absurdly mistake. The human infections of Diphyllobothrium latum,Toxocara canis,and Triodontophorus minor discovered by Lin(1924) from Beijing were the first records in the country. A doubtful malaria case reported from Chongqing by Hung(1944) should be revised as the first case of babesiosis in China. The above-presented examples suggest that the truthful record of parasitic pathogens is an important base for the discovery history of parasitic diseases. With comments on the research progress of human parasitic diseases in different historical stages,it seems that the trends of medical parasitology development in China have been synchronous with the research activities in the area.
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Using scanning electromicroscope, seven species of anopheline mosquito eggs were studied. Among them, the microstructure of exochorion of Anopheles liangshanensis. An. kweiyangensis An. kunmingensis, An. hyrcanus and An. messeae were not reported before. The results showed that ultrastructure of plastron network, frill and tubercles of deck were useful in distinguishing sibling species of anopheline eggs. Microstructure of float had little difference between species. The micropylar area and lobed tubercles presented obvious intraspecific variations, so, it should be careful for using in classification.
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Two isolates of Plasmodium jalcipanm from Hainan Province were cloned by limiting dilution method and eight clones were established. Some characteristics including drug sensitivity and antigenicity of these clones were observed. The results showed that the established clones were different from each other in chloroquine sensitivity and antigenicity. The ID50 of chloroquin3e against 6 clones from isolate Fcc-7801 varied between 60.60 and 13.08 nmol/L The ID50 of chloroquine against 2 clones from isolate Fcc-1 were 93.63 and 49.64 nmol/L, respectively. According to the indirect fluorescent antibody (IFA) Teactivity of these clones with a panal of murine anti-gpl95 McAbs, 8 clones could be divided into 5-serotypes, 3 of which were consistent with the Ⅴ , Ⅵ, Ⅶ serotypes devided by J. S. McBride (1985).
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1.00) than the subgenus Cellia ( An.kochi.An.maculanis, An.dirus. An. jeyporiensis, and An.mininus) with Rm value
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The zymograms of EST in six species of anopheline mosquitoes (Female) were studied with polyacrylamide gel electrophoresis.The zymograms of EST were divided into three groups,EST Ⅰ, EST Ⅱ and EST Ⅲ, respectively.The Rm of subgenus Anopheles (including An sinensis,An.anthropophagus and An.kunmigensis) was larger than that of subgenus Cellia (including An.dirus, An.mmmus and An.kochi) in main bands of EST Ⅲ. The results showed specific characteristics at the taxonomic level of subgenus. The six species of anopheline mosquitoes could be differentiated from each other by the particular band pattern and Rm in EST n groups.
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Mitotic chromosomal karyotypes and their heterochromatin banding of thirteen anopheline mosquitoes in China were observed. Five species belonging to subgenus Cellia were Anopheles maculatus, An. dints, An. kochi, An. splendidus and An. minimus. Eight species belonging to subgenus Anopheles were An. barbirostris, An. messeae, An. crawfordi, An. kunmingensis, An. kweiyangensis, An, hyrcanus, An. sinensis and An. anthropophagus. The results show that the interspecific differences of sex chromosomes and heterochromatin differences in autosomes are useful in anopheline sibling species identification. Two types of completely different chromosomal karyotypes of An. maculatus were found from Yunnan and Sichuan Provinces, and two types of obviously different heterochromatin banding of An. dims were found from Hainan and Yunnan Provinces. Thus, An. maculatus and An. dirus are respectively sibling species complex in China. The salivary gland polytene chromosomes of An. hyrcanus, An. sinensis and An. anthropophagus were also studied. The authors find that the main differ ences of polytene chromosomes of the three species are fixed paracentric inversions in arm 2L.