Limulus Factor C, a serine protease zymogen from the amoebocytes of the limulus, has high affinity for endotoxin. When Factor C is activated by endotoxin, it hydrolyses artificial tripeptide substrate and measurable products are released, so it can be used as an alternative reagent for endotoxin analysis. Factor C gene of Tachypleus tridentatus was obtained through RT-PCR and the recombinant protein was expressed by Bac-to-Bac/BmNPV baculovirus expression system in silkworm larvae. The activity of Factor C was detected with diluted serum of silkworm larvae, and the sensitivity of endotoxin detected was 0.2 EU/mL when the serum was diluted at 1:500. The silkworm larvae expressed limulus Factor C could be used to develop a new low-cost endotoxin test reagent.
Animals ; Arthropod Proteins ; biosynthesis ; Baculoviridae ; Bombyx ; metabolism ; Enzyme Precursors ; biosynthesis ; Genetic Vectors ; Larva ; metabolism ; Recombinant Proteins ; biosynthesis ; Serine Endopeptidases ; biosynthesis

Rickettsial infections (Rickettsioses) are the causes of acute fever found in Thailand. It is classified as acute febrile illnesses transmitted by bloodsucking arthropod vectors (tick, flea, and chigger). This research investigated pathogens of scrub typhus in vectors from Bangkaew District, Phatthalung Province. A total of 303 pools of vector samples were ticks (Rhipicephalus sanguineus, R. microplus, and Haemaphysalis sp.), fleas (Ctenocephalides felis orientis, C. f. felis, and C. canis), and chiggers (Leptotrombidium deliense, Aschoschoengastia indica, Blankaartia acuscutellaris and Walchia disparunguis pingue) collected from reservoir hosts (dogs and rodents). The 17 and 56 kDa gene of Rickettsia causing scrub typhus were found in 29% of ticks and 98% of flea. DNA sequence analysis reveeled the detected strains were R. asembonensis and Rickettsia sp. cf1 and 5.The chiggers, 1%, were infected with Rickettsia strain TA763, a pathogen of scrub typhus.
Animals ; Arthropod Vectors ; Cats ; Felis ; Fever ; Orientia tsutsugamushi ; Rickettsia ; Scrub Typhus ; Sequence Analysis, DNA ; Siphonaptera ; Thailand ; Ticks ; Trombiculidae

A survey was performed in Korea to monitor the prevalence of five bovine arboviruses [Akabane virus, Aino virus, Chuzan virus, bovine ephemeral fever (BEF) virus, and Ibaraki virus] in arthropod vectors, such as Culicoides species. To determine the possible applications of survey data in annual monitoring and warning systems in Korea, we examined the prevalence of bovine arboviruses in arthropod vectors using RT-PCR. To compare the sensitivity and specificity of virus detection, nested PCR was also performed in parallel for all five viruses. Using the RT-PCR, the detection limits were at least up to 10(1.5), 10(2.8), 10(2.0), 10(1.8), and 10(4.0) TCID50/ml for Akabane virus, Aino virus, Chuzan virus, BEF virus, and Ibaraki virus, respectively. When nested PCR was performed using 1 micronl of PCR product, the detection limits were increased, to 10(0.05), 10(1.8), 10(1.0), 10(0.008), and 10(2.0) TCID50/ml for Akabane virus, Aino virus, Chuzan virus, BEF virus, and Ibaraki virus, respectively. Thus, nested PCR increased the sensitivity of the virus detection limit by 1~2 log. We pooled 30~40 mosquitoes in one sample. We collected 113 samples in 2006, 135 samples in 2007, and 100 samples in 2008. Among these samples, Chuzan virus and BEF virus genes were detected at a range between 0.82% and 1.19%, and Akabane virus, Aino virus, and Ibaraki virus genes were detected at less than 0.20%. These data may provide some insight into future epidemiological studies of bovine arboviral diseases in Korea.
Animals ; Arboviruses ; Arthropod Vectors ; Arthropods ; Cattle ; Ceratopogonidae ; Culicidae ; Ephemeral Fever ; Epidemiologic Studies ; Korea ; Limit of Detection ; Organothiophosphorus Compounds ; Palyam Virus ; Polymerase Chain Reaction ; Prevalence ; Sensitivity and Specificity ; Viruses

Dung beetles (family Scarabaeidae) are one of the largest families of beetles worldwide. Due to biological behavior of these arthropods, they are considered to play an important role in the life cycle of some helminths. In the present study, dung beetles collected from cattle pastures in rural areas of Ardabil province, north-west of Iran were examined for infection with larval stages of helminths. According to the results, nematodes of 2 genera were identified including Rhabditis and Gongylonema. The more common species was Rhabditis sp. which was found in 9 species of beetles. Out of 15 different species of dung beetles, Copris lunaris was the only scarabaeid to be found naturally infected with the larval stages of Gongylonema sp. Our new findings introduce C. lunaris as a potential biological vector for transmission of Gongylonema sp. to vertebrates in the surveyed region.
Animals ; Arthropod Vectors/*parasitology ; Beetles/*parasitology ; Humans ; Iran/epidemiology ; Larva/physiology ; Rhabditoidea/isolation & purification/physiology ; Spirurida Infections/epidemiology/parasitology/*transmission ; Spiruroidea/isolation & purification/*physiology

In this review, we mainly focus on zoonotic encephalitides caused by arthropod-borne viruses (arboviruses) of the families Flaviviridae (genus Flavivirus) and Togaviridae (genus Alphavirus) that are important in both humans and domestic animals. Specifically, we will focus on alphaviruses (Eastern equine encephalitis virus, Western equine encephalitis virus, Venezuelan equine encephalitis virus) and flaviviruses (Japanese encephalitis virus and West Nile virus). Most of these viruses were originally found in tropical regions such as Africa and South America or in some regions in Asia. However, they have dispersed widely and currently cause diseases around the world. Global warming, increasing urbanization and population size in tropical regions, faster transportation and rapid spread of arthropod vectors contribute in continuous spreading of arboviruses into new geographic areas causing reemerging or resurging diseases. Most of the reemerging arboviruses also have emerged as zoonotic disease agents and created major public health issues and disease epidemics.
Africa ; Alphavirus* ; Animals, Domestic ; Arboviruses* ; Arthropod Vectors ; Asia ; Encephalitis ; Encephalitis Virus, Venezuelan Equine ; Encephalitis Virus, Western Equine ; Encephalitis Viruses ; Encephalomyelitis, Equine ; Epidemiology* ; Flaviviridae ; Flavivirus* ; Global Warming ; Humans ; Population Density ; Public Health ; South America ; Togaviridae ; Transportation ; Urbanization ; Zoonoses

Attempts were made to isolate and identify Korean bovine Babesia parasite. Blood samples were collected from Holstein cows in Korea, and Babesia parasites were propagated in SCID mice with circulating bovine red blood cells for isolation. The isolate was then antigenically and genotypically compared with several Japanese isolates. The Korean parasite was found to be nearly identical to the Oshima strain isolated from Japanese cattle, which was recently designated as Babesia ovata oshimensis n. var. Haemaphysalis longicornis was the most probable tick species that transmited the parasite.
Animals ; Arthropod Vectors/parasitology ; Babesia bovis/genetics/*isolation & purification ; Babesiosis/parasitology ; Base Sequence ; Cattle/*parasitology ; Cattle Diseases/parasitology ; DNA, Protozoan/genetics ; DNA, Ribosomal/genetics ; Erythrocytes/parasitology ; Korea ; Mice ; *Mice, SCID ; Molecular Sequence Data ; Phylogeny ; Ticks/parasitology

OBJECTIVETo construct a vector encoding T-cell epitopes of major allergen group 1 of Dermatophagoides pteronyssinus as a vaccine delivered by MHC class II pathway.

METHODSThe nucleotide sequences of the 3 target genes were synthesized, including TAT, IhC and the recombinant fragment of Der p 1 encoding 3 T-cell epitopes. After amplification of the 3 target fragments by PCR and digestion with corresponding restriction endonucleases, the recombinant gene TAT-IhC-Der p 1-3T was ligated using T4 DNA ligase and inserted into the prokaryotic expression vector pET28a(+) to construct the recombinant plasmid pET-28a(+)-TAT-IhC-Der p 1-3T, which was confirmed by digestion with restriction endonucleases and sequencing. The recombinant vector was transformed into E. coli strain BL21 (DE3) and induced with IPTG, and the induced protein TAT-IhC-Der p 1-3T was detected by SDS-PAGE. After purification, the recombinant protein was confirmed by Western blotting and its allergenicity tested using IgE-binding assay.

RESULTSThe recombinant plasmid pET-28a-TAT-IhC-Der p 1-3T was successfully constructed as confirmed by restriction endonuclease digestion and sequencing and the expression of the recombinant protein TAT-IhC-Der p 1-3T was induced in E. coli. Western blotting verified successfull purification of the target protein, which showed a stronger IgE-binding ability than Der p 1.

CONCLUSIONWe successfully constructed a recombinant expression vector pET-28a-TAT-IhC-Der p 1-3T expressing a T-cell epitope vaccine delivered by MHC II pathway with strong IgE-binding ability, which provides a basis for further study on specific immunotherapy via MHC class II pathway.

Allergens ; immunology ; Animals ; Antigens, Dermatophagoides ; immunology ; Arthropod Proteins ; immunology ; Base Sequence ; Cloning, Molecular ; Cysteine Endopeptidases ; immunology ; Dermatophagoides pteronyssinus ; Epitopes, T-Lymphocyte ; Escherichia coli ; Gene Expression ; Genes, MHC Class II ; Genetic Vectors ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; Vaccines ; immunology

A total of 9,281 larval chigger mites were collected from small mammals captured at Hwaseong-gun, Gyeonggi-do (Province) (2,754 mites from 30 small mammals), Asan city, Chungcheongnam-do (3,358 mites from 48 mammals), and Jangseong-gun, Jeollanam-do (3,169 for 62 mammals) from April-November 2009 in the Republic of Korea (= Korea) and were identified to species. Leptotrombidium pallidum was the predominant species in Hwaseong (95.8%) and Asan (61.2%), while Leptotrombidium scutellare was the predominant species collected from Jangseong (80.1%). Overall, larval chigger mite indices decreased from April (27.3) to June (4.9), then increased in September (95.2) and to a high level in November (169.3). These data suggest that L. pallidum and L. scutellare are the primary vectors of scrub typhus throughout their range in Korea. While other species of larval chigger mites were also collected with some implications in the transmission of Orientia tsutsugamushi, they only accounted for 11.2% of all larval chigger mites collected from small mammals.
Animals ; Arachnid Vectors ; Larva/*microbiology ; Orientia tsutsugamushi/*isolation & purification ; Republic of Korea ; Rodentia ; Scrub Typhus/*microbiology ; Trombiculidae/*classification/*microbiology

OBJECTIVETo explore the fact that the east border of Heilongjiang had been a lyme disease natural focus,we investigated the species and distribution of ticks and isolated bacteria from ticks and identified genomic species of Borrelia burdorferi sensu lato. This study provided evidence for prevention and control of lyme disease.

METHODSTicks were caught by flagging method and Direct immunofluorescence method was used to detect the rate of bacteria borne by the tick. BSK UI culture medium was used to isolate the agent and Specific McAbs were used to identify the bacteria. SDS-PAGE protein profile and PCR-RFLP method were also used to identify the species of Spirochetes.

RESULTSTicks, collected from China-Russia border of east Heilongiiang province were classified including Ixodes persulcatus Schulze, Dermacentor sivarum Olener, Haemaphysalis concinna Kock,and Haemaphysalis japonica Kock. We found that the distributon of ticks was different under different circumstances and the predominant species were also different in different ports. The rate of bacteria borne by Iodes persulaatus Schulze was 31.4% ,by Dermacentor sivarum Olener and Haemaphysalis concinna Kock were 2.2% and 3.8%, respectively. However,it was negative for Haenaphysalis japonica Kock. Spirochetes isolated from Ixodes persulcatus Schulze were collected from Dongning and Tongjiang while Genomic species of Spirochetes, isolated from ticks of the border belonged to B. garinii.

CONCLUSIONAll the results showed that the east border of Heilongjiang province was the natural focus of lyme disease.

Animals ; Arachnid Vectors ; classification ; microbiology ; Borrelia burgdorferi ; classification ; genetics ; isolation & purification ; China ; Humans ; Lyme Disease ; microbiology ; Russia ; Ticks ; classification ; microbiology

Deer serve as reservoirs of tick-borne pathogens that impact on medical and veterinary health worldwide. In the Republic of Korea, the population of Korean water deer (KWD, Hydropotes inermis argyropus) has greatly increased from 1982 to 2011, in part, as a result of reforestation programs established following the Korean War when much of the land was barren of trees. Eighty seven Haemaphysalis flava, 228 Haemaphysalis longicornis, 8 Ixodes nipponensis, and 40 Ixodes persulcatus (21 larvae, 114 nymphs, and 228 adults) were collected from 27 out of 70 KWD. A total of 89/363 ticks (266 pools, 24.5% minimum infection rate) and 5 (1.4%) fed ticks were positive for Anaplasma phagocytophilum using nested PCR targeting the 16S rRNA and groEL genes, respectively. The 16S rRNA gene fragment sequences of 88/89 (98.9%) of positive samples for A. phagocytophilum corresponded to previously described gene sequences from KWD spleen tissues. The 16S rRNA gene fragment sequences of 20/363 (5.5%) of the ticks were positive for A. bovis and were identical to previously reported sequences. Using the ITS specific nested PCR, 11/363 (3.0%) of the ticks were positive for Bartonella spp. This is the first report of Anaplasma and Bartonella spp. detected in ticks collected from KWD, suggesting that ticks are vectors of Anaplasma and Bartonella spp. between reservoir hosts in natural surroundings.
Anaplasma/genetics/*physiology ; Animals ; Arachnid Vectors/microbiology ; Bartonella/genetics/*physiology ; Chaperonin 60/genetics ; Deer/parasitology ; Disease Reservoirs/veterinary ; RNA, Ribosomal, 16S/genetics ; Republic of Korea/epidemiology ; Ticks/*microbiology