Babesia microti, a causative agent of human babesiosis, a tick-borne zoonotic disease, has been detected in Ixodes persulcatus ticks in Mongolia. Large-scale surveys of B. microti in ticks and its infectivity in rodents are essential. Of 458 I. persulcatus ticks, 94 females and 176 males were screened for B. microti infection, while 111 female and 77 male ticks were used to investigate B. microti transmission to BALB/c mice and hamsters. The prevalence of B. microti was higher in female ticks (26.6%) than in males (13.1%), with nearly one-sixth (17.8%) of ticks carrying the parasite. The infectivity of Mongolian isolates, as a sister clade to the U.S. lineage, remains unclear, as they failed to infect rodents. Our study suggests that the risk of B. microti transmission is approximately of one-sixth of I. persulcatus ticks, with a higher risk observed in females than in males.

Babesia microti, a haemoprotozoan parasite, is transmitted to humans by the bites of Ixodes species ticks, and following the parasite infection is more frequently transmitted via parasite-unscreened donations than parasite-screened donations. In Mongolia, B. microti infection has been reported in humans and Ixodes persulcatus ticks, but the B. microti transmission risk has not been investigated yet in blood donations. In this study, we verified B. microti-free status in blood donations stored in blood transfusion local services, especially in the I. persulcatus-endemic areas using newly designed 18S ribosomal gene (18S rRNA) specific PCR primers.The newly designed first PCR primers bind conserved region of 18S rRNA sequences of B. microti, Babesia divergens and Babesia venatorum, whereas they have at least 2 mismatching single nucleotide polymorphisms to Ixodes species and Homo sapiens 18S rRNA sequences. While a set of second PCR primers also binds conserved region of B. microti 18S rRNA sequences, whereas the second PCR primers have at least 5 mismatching single nucleotide polymorphisms to B. divergens and B. venatorum 18S rRNA sequences. Newly designed primers have a sensitivity as high as that of routinely used primers. Using these primers, we have verified parasite-free status in blood donations to reduce the B. microti transmission risk in the I. persulcatus-endemic areas in Mongolia. The monitoring of the B. microti transmission risk should be especially continued in the I. persulcatus-endemic areas due to unscreened blood donations are often associated with higher pathogen transmission risk than screened blood donations.

Cryptosporidium is a small protozoan parasite that infects the microvillous region of epithelial cells in the digestive and respiratory tract of vertebrates. It is an obligate intracellular parasite of man and other mammals, birds, reptiles and fish. We were purification Cryptosporidium spp. oocysts with saturated sugar solution from diarrhea cattle. And experimental infection use by control C. parvum (Germany) strain and Cryptosporidium spp. with oral 1x107 oocyst/ml doses in per BALB/c mouse. Post inoculation detected Cryptosporidium oocysts within a week in the feces mice and counted average Cryptosporidium spp. (Mongolia) 1.033±0.2x106 (p<0.025) and C. parvum 0.9±0.05x106 (p<0.001) oocyst/ml each other period of the experiment. Mice from the control and experimental trials were slaughtered on the 30th day post inoculation and examination for pathology. Pathology examination, the asexual and sexual developmental stages of Cryptosporidium were evident in the gastric and small intestinal epithelium of the experimental mice, in oocysts, in the glandular layer of gastric submucosa, and in the microvillous of small intestinal wall submucosa. In the future, we will do focus of molecular biological and isolated Mongolian laboratory strain of Cryptosporidium spp.. Cryptosporidium spp. үүсгэгчийг BALB/c хулганад зориудын халдвар хийж туршсан дүн Криптоспорид бүх төрлийн сээр нуруутан амьтдын ходоод, нарийн гэдэсний хучуур эсэд шимэгчилдэг эгэл биетэн. Бид усархаг чацга алдсан үхрийн баасны сорьцоос криптоспоридын ооцистыг сахарын ханасан уусмалаар цэвэршүүлэн авлаа. Монгол дахь уг үүсгэгчийг 1х107 ооцист/мл тунгаар BALB/c үүлдрийн хулганад зориудын халдвар хийж, хяналтаар С. parvum (Герман) омгийг ашиглав. Халдвар хийснээс хойш 7 хоногийн дараагаас эхлэн туршилтын бүлэг тус бүрийн хулганы баасаар криптоспоридын ооцист ялгарч, туршилтын нийт хугацаанд дунджаар монгол дахь криптоспоридын үүсгэгч 1.033±0.2х106 (p<0.025), C. parvum зүйлийн 0.9±0.05x106 (p<0.001) ооцист/мл тоологдлоо. Хяналт, туршилтын бүлгийн хулганыг халдвар хийснээс хойш 30 дахь хоногт нядалж эмгэг бүтцийн өөрчлөлтийг судлав. Туршилтын бүлгийн хулганы ходоод, нарийн гэдэсний ханын хучуур эсүүд гэмтэж, үүсгэгчийн ооцист ихээхэн халдварлаж, ходоодны ханын салст бүрхүүлийн булчирхайлаг эдийн давхаргад байршин шимэгчилжээ. Нарийн гэдэсний бичил цэлмэнгийн ёроол хэсгээр криптоспоридын бэлгийн бус, бэлгийн хөгжлийн үе шатууд тод илрэв. Бид хоруу чанар өндөр Cryptosporidium spp.-ийн лабораторийн омог гарган авахаар ажиллаж байна. Түлхүүр үг: C. parvum омог, баасны сорьц, ооцист, үхэр, Гематоксилин-Эозин будаг, сахарын ханасан уусмал

During the study for biodiversity of wild mammals and their parasite and infectious agents, there have more risky to get natural hotspots due to global warming, huge number of domesticated animals and deficiency of pasture in Mongolia. Therefore, a major concern of researchers and rangers is survey to ascertain parasites of wild animals, particularly parasitism of migratory birds. We used to the study totally 224 feacal samples of wild animals, including 96 wild sheep, 101 ibex, 53 wild goose and 24 wild horse. The study results, found by helminth fauna of 7 genera such as Paramphistomum spp, Moniezia spp, Neoascaris sрp, Trichostrongylus spр, Nematodirus sрp, Haemonchus spp, Trichuris spp, belonging to 3 main classes of helminthes and Coccidia spp protozoa in feacal samples of mountain sheep and the 5 genera of helminthes such as Paramphistomum spp, Moniezia spp, Neoascaris sрp, Nematodirus sрp, Trichuris spp and protozoa Coccidia spp in feacal samples of ibex. The Paramphistomum spp have been found in mountain sheep and ibex for the first time in Mongolia. The floatation method of feacal egg determination, shown that nematode infection of Ascaridae and Strongylidae in wild horses. The egg of Capillaria spp and oocyst of Eimeria spp in feces of swan goose and the eggs of two genera of nematode (Ascaridae spp, Capillaria spp) and oocyst of Eimeria spp in feces of grey goose were found in the study. The parasite fauna in feacal samples of certain wild host animals was considered by the prevalence of infection in Mongolia. Зэрлэг амьтдын гельминтийн судалгаа Цаг агаарын дулаарал, мал сүргийн тооны өсөлт, бэлчээрийн хомсдол, хүний хүчин зүйл зэргийн улмаас мал, амьтны паразитын тархалт нэмэгдэж, улмаар зэрлэг амьтад гадна, дотор паразит, халдварт болон өвчнөөр өвчлөн байгалийн голомт үүсгэх явдал гарсаар байна. Иймд зэрлэг амьтад, нүүдлийн шувуудын паразит өвчний халдварлалтыг судлан тогтоох, түүнтэй тэмцэх, сэргийлэх арга боловсруулах нь судлаачид, байгаль хамгаалагчдын өмнө тулгамдсан асуудал болжээ. Бид судалгаандаа 96 толгой аргаль, 101 толгой янгир, 53 толгой галуу, 24 толгой тахийн баасны сорьцыг цуглуулж шинжилгээ хийхэд аргальд трематод, цестод, нематодын 3 ангийн Paramphistomum spp, Moniezia spp, Neoascaris sрp, Trichostrongylus spр, Nematodirus sрp, Haemonchus spp, Trichuris spp зэрэг 7 төрлийн гельминт, Coccidia spp төрлийн эгэл биетэн, янгирт Paramphistomum spp, Moniezia spp, Neoascaris sрp, Nematodirus sрp, Trichuris spp зэрэг 5 төрлийн гельминтүүд, Coccidia spp төрлийн эгэл биетэн илэрч тодорхойлогдоод байна. Үүнээс Paramphistomum spp төрлийн трематодыг аргаль, янгираас анх удаа илрүүлсэн болно. Харин тахь нь Ascaridae, Strongylidae төрлийн нематодоор халдварласан болох нь тогтоогдов. Хошуу галууны баасны сорьцноос Capillaria spp төрлийн нематодын өндөг, Eimeria spp төрлийн эгэл биетний ооцистын халдвар илэрсэн бол бор галууны нийт баасны сорьцноос 2 төрлийн нематод (Ascaridae spp, Capillaria spp)-ын өндөг, Eimeria spp төрлийн эгэл биетний халдвар илрэв. Энэ судалгаагаар Монгол орны аргаль, янгир, тахь, нүүдлийн шувууд ямар төрөл, зүйлийн паразитаар халдварласан ерөнхий дүр зураг буй болов. Түлхүүр үг: Аргаль, янгир, тахь, нүүдлийн шувуу, баасны сорьц, гельминт, өндөг, ооцист

Demodicosis, also called demodectic mange infests human and animals worldwide. It is a skin parasitic disease caused by demodex mites. Of 615 goats randomly selected from Central and Western regions of Mongolia were carried out to detect the demodectic mange by clinical examination. The prevalence of Demodicosis was 1.3% in goats. It was considered as suspected /diseased that the skin had multifocal nodules with grayish, dried rough surface which were approximately 3-9 mm in diameter. The encrusted lesions of skin, especially over face, necks, shoulder blades, groin and hair loss were found on selected Mongolian goats. The samples of skin scrapings from the goats with above clinical signs were collected. The mites were identified as Demodex spp by parasitological examination. DNA was extracted from positive samples. The specific genes (mt16s rDNA) of Demodex spp. was detected by PCR. The 16s rDNA fragment was sequenced and aligned. The presence of D.-caprae in Mongolian goats is confirmed by molecular biological

Sika deer (Cervus nippon) is widely distributed in Asian countries and is one of the most common wildlife animals in Hokkaido, Japan. Previous studies identified Theileria spp. in sika deer in Japan including Theileria sp. Thrivae belonging to T. cervi group and Theileria sp. sola belonging to T. capreoli group. However, the studies failed to differentiate these two species without sequencing. Therefore, epidemiological information on cervine theileriosis in Hokkaido, Japan is limited. This study differentiated the two Theileria spp. using restriction fragments length polymorphism (RFLP). Based on the PCR-RFLP, Theileria spp. were identified in 103 (88.0%) of 117 samples, and the prevalence of each parasites were 86.3% (n = 101) and 57.3% (n = 67) for Theileria sp. Thrivae and T. capreoli-like, respectively. Phylogenetic analysis based on the 18S rRNA showed a close relationship between Theileria sp. Thrivae and T. cervi in China. In addition, phylogenetic analysis of internal transcribed spacer regions also showed a close relationship between Theileria sp. Thrivae and T. cervi.

Objective: The purpose of this study was to identify cases of human echinococcosis in the Bayan-Ulgii, Khovd provinces, to determine their WHO classification by ultrasound and to identify the haplotype in surgically cysts removed. Methods: A total of 1063 people participated in ultrasound screening. Patients who had a positive echinococcosis by ultrasound underwent serologic testing for detection of antibodies for echinococcosis. Results: Ninety people were positive for the liver cystic lesions by ultrasound screening of which 12 cases were newly diagnosed cystic echinococcosis. Four were cystic echinococcosis stages 1-2, five were stage 3, while three were cystic echinococcosis stage 4. Thirty-three patients with previously known echinococcosis stage 2 and alveolar echinococcosis stage 4 were also examined. Out of 29 cystic echinococcosis cases, 24 underwent treatment. Of 24 who underwent treatment, only one was cured. The haplotypes E. canadensis G6/7 and E. granulossus s.s. G1 were confirmed by evaluating the cytochrome c oxidase subunit I gene of mitochondrial DNA in two patients in Bayan-Ulgii. Conclusion: Ninety people were positive for cystic echinococcosis in western Mongolia of which 12 cases were newly diagnosed cases. Of the 12 new cystic echinococcosis cases, 4 were stages 1-2, 5 were stage 3 and 3 were stage 4 by WHO classification. The haplotype in Bayan-Ulgii population were E. canadensis and s.s.

Transparency and accountability initiatives have emerged as a potential solution to combat corruption and increase public benefits from the extractive sector in resource-abundant countries. The Extractive Industries Transparency Initiative (EITI) is one such initiative, through which 49 resource-rich countries have disclosed a cumulative 282 fiscal years of government revenues amounting to US$1.9 trillion since 2003. This paper explores the potential for promised benefits of increased disclosure to be realized, in the form of improved resource governance. Building on the social accountability literature, a framework is proposed and then applied to the Mongolian context to examine which stages of the framework work well, and which fail to perform. Two types of contracts are analyzed, water usage agreements and community benefit-sharing agreements. Although Mongolia is recognized as a leading performer by international EITI standards, the analysis concludes that the framework’s latter stages from disclosure to improved resource governance are incomplete. The policy implication is that greater attention to mobilization and citizen empowerment is needed to ensure that contract transparency can meaningfully contribute towards improved governance.

The water buffalo industry is a vital part of the Philippine livestock economy and is an essential contributor to the developing local dairy industry. Although relatively less susceptible to diseases, water buffaloes can still be infected and can act as reservoirs of tick-borne pathogens (TBPs). However, limited information is available regarding the prevalence of tick-borne infections in water buffaloes in the Philippines. This study was conducted to identify TBPs harbored by water buffaloes and to characterize these pathogens molecularly. One hundred water buffalo blood samples collected from three areas in Bohol, Visayas region, Philippines were screened for various TBPs using pathogen-specific PCR assays. TBPs were detected in 46% of the samples (39% singly infected, 7% coinfected). The pathogens detected were Anaplasma marginale (29%), Babesia bovis (21%), and B. bigemina (3%). None of the blood samples were positive for Theileria annulata, T. orientalis, and B. ovata. A. marginale infection rates were significantly higher (37.5%) among water buffaloes aged <= 6 years (P = 0.046) than those > 6 years old (18.2%) and was detected only in Bulgarian Murrah (36.1%) and US Murrah (25.9%) breeds. Phylogenetic analyses revealed that groEL sequences of A. marginale were 100% identical with isolates from the Philippines (Batangas and Cebu) and China. Two B. bigemina RAP-la gene sequences were identical to each other and were homologous with previous isolates from Thailand, Indonesia, Uruguay, and the Philippines. Moreover, four B. bovis SBP-2 partial sequences obtained in this study had 92.4-99.7% identities. This study is the first molecular detection and characterization of A. marginale, B. bigemina and B. bovis in water buffaloes in the Visayas region, and the first molecular confirmation of B. bovis infection in water buffaloes in the country. The findings presented in this study may serve as baseline data for crafting effective tick-borne disease surveillance and prevention programs in Bohol and in the Philippines.

Gold-headed tick, Dermacentor nuttalli is widely distributed in Mongolia and an important vector of transovarially transmitting pathogens to domestic animals. In this study, we describe the morphology of the ovary, as well as the process of the vitellogenesis and ovigenesis on oocytes of the tick, Dermacentor nuttalli. The oocytes were classified into stages that varied from I to V, according to: cytoplasm appearance, presence of the germ vesicle, position, development, presence of yolk granules, accumulation and presence of chorion. To our knowledge, this study is first description on oocytes development stages evaluated by Balashov (1964) of D. nuttalli female and its vitellogenesis. Dermacentor nuttalli хачгийн ооцитын хөгжил Хураангуй: Цус бүрэн сорж, эвцэлдэж хувалзалсан Dermacentor nuttalli эмэгчин хачгийн ооцит, өндөгний хөгжлийг судалснаар хачгийн физиологи, биологийн онцлогийг тогтоох нь шинжлэх ухаан, онол, практикийн өндөр ач холбогдолтой юм. Энэ судалгаагаар D. nuttalli хачгийн ооцитын вителлогенез, овигенез, өндгөвчний хэлбэр зүйн хувирлыг судлан тогтоолоо. Хөгжлийн янз бүрийн шатаар хөгжиж буй ооцитуудыг цитоплазмын илрэл, герминал мөхлөгт цэврүүнцрийн илрэл, байрлал, хөгжил, уургийн мөхлөгүүдийн илрэл, хуримтлагдалт, цэлмэнт бүрхүүлийн үүсэл, хөгжлийн явцаар нь 5 шат болгон ангилав (I-V). Энэхүү судалгаа нь D. nuttalli хачгийн ооцитын хөгжлийг Ю.С.Балашов (1964)-ын үнэлгээгээр үнэлж, хөгжлийн үе шатуудыг тогтоосон анхны судалгаа юм. Түлхүүр үг: Өндгөвч, овигенез, Балашовын ангилал, вителлогенез

The blood sucking 4 species ticks of Haemaphysalis genus were distributed in Mongolia, that an important vectors of diseases transmitting to human and animals and causing economic lost. Newly found Haemaphysalis females in Selenge province was confirmed to H. concinna species based on following features: strong lateral margins of palp article II, heart-shaped scutum with widest at middle, short-wide hypostome, sharpen cornua of basis capituli, posterior margins of basis capituli, ventral surface of basis capituli, latero-anterial I festoon connected with lateral, anterior groove of genital pore and additional spur of leg I trochanter. To confirm morphological description of H. concinna, CoxI sequence analysis was done and results was showed H. concinna CoxI sequences in GenBank with 86-89% identity. The results indicate that the Haemaphysalis concinna tick was distributed in Mongolia and it is announcing that the spread of Haemaphysalis tick in Selenge aimag. Сэлэнгэ аймгийн нутгаас шинээр олдсон Heamphysalis concinna хачгийн хэлбэр зүйн судалгаа Хураангуй: Монгол оронд Haemaphysalis төрлийн 4 зүйлийн хачиг тархан хүн, мал амьтанд шимэгчилж өвчин дамжуулан хохирол учруулдаг. Сэлэнгэ аймгийн нутгаас олдсон хачиг нь хэлбэр зүйн ерөнхий онцлогоороо Haemaphysalis төрөлд хамаарагдаж байлаа. Хошууны пальпын II үеийн хажуугийн нэмэлт хурц ирмэг, бамбайн дунд хэсгээрээ өргөссөн зүрх мэт хэлбэр, богино өргөн гипостом, хошууны суурийн булангийн эвэр, арын ирмэгийн илрэлт, хошууны суурийн доод гадаргуугийн хэлбэр, хачгийн захын I фестон нь зөвхөн биеийн хажуугийн ховилтой холбогдсон байдал, бэлгийн сүвийн урьдах зураасны онцлог, бэлгийн сүвийн хэлбэрийн онцлог, ялгадасны сүвийн арын ховилын онцлог, I хөлийн I үеийн нэмэлт өргөс хэлбэрийн өргөсөлт зэрэг онцлогоороо Haemaphysalis concinna зүйлийн хачиг болох нь батлагдав. Энэхүү хачгийн морфологийн ялган танилтыг CохI генийн нуклеотидын дарааллыг тогтоон баталгаажуулахад ГенБанкинд бүртгэлтэй H. concinna хачгийн нуклеотидын дарааллуудтай 86-89%-ийн адил байлаа. Бидний судалгааны үр дүнд хэлбэр зүйн онцлог, нуклеотидын дараалал тогтоох шинжилгээгээр Монгол оронд H. concinna зүйл хачиг тархсан нь тогтоогдсон тул Монгол орны хачгийн жагсаалтанд оруулж болохыг баталгаажуулж байна. Мөн энэхүү судалгаагаар Сэлэнгэ аймгийн нутгаас Haemaphysalis төрлийн хачиг анх удаа илрүүлэв. Түлхүүр үг: Цус сорогч хачиг, Haemaphysalis concinna, хэлбэр зүй, Сэлэнгэ аймаг

The reindeer Rangifer tarandus Linnaeus, 1758, is a local animal (west) taiga in Tsagaannuur soum of Khuvsgul aimag. The species, one of semi-domesticated populations, is well adapted to harsh climate as like as tundra and sub tundra areas. The reindeer was slaughtered by herdsmen for their own food consumption and the post-mortem examination was done by incomplete methods of necropsy (Skrybini, 1945). It is a first time detection to find for Thysaniezia spp. tapeworm in reindeer from Mongolia. In the future we will expand the study of parasitic and infectious diseases of the reindeer. Баруун тайгын цаа буганд thyzaniezia төрлийн туузан хорхой илрүүлсэн дүн Хураангуй: Хөвсгөл аймгийн Цагааннуур сумын Баруун тайгад Rangifer tarandus, Linnaeus, 1758 зүйлийн цаа буга (буган цаа) өргөн тархжээ. Цаа буга нь тундр, тундр орчмын бүсийн эрс тэс уур амьсгалд дасан зохицсон хагас гэршсэн хөхтөн юм. Дээрх бүс нутгаас хүнсний хэрэгцээнд нядласан цаа буганд цагаан хорхойн судлалын бүрэн бус задлан шинжилгээг хийв. Манай оронд маллагдаж буй цаа буганаас анх тутам Thysaniezia төрлийн туузан хорхойг илрүүлэв. Цаашид цаа бугын зүй бус хорогдлын үндсэн шалтгаан паразиттах, халдварт, халдваргүй өвчний судалгааг өргөжүүлэх чухал шаардлагатай. Түлхүүр үг: Нарийн гэдэс, задлан шинжилгээ, Rangifer tarandus, баасны дээж, хавтгай хорхой

The survey on the Amur ide, Leuciscus waleckii, conducted between May to October, 2015 along Kherlen River in Khentii province, Mongolia. Ichthyoxenus amurensis was found in the body cavity of the Amur ide. A total of 12 individuals of Ichthyoxenus amurensis were collected from eight of 25 fish specimens (prevalence=32.0%). In the body cavity of the Amur ide, Leuciscus waleckii was found within a thin-walled membranous sac, an orifice near the ventral region of the pectoral fin. Ichthyoxenus amurensis caused deleterious effects in fish, such as decreases in mean weight and growth. This is the first report about detecting of Ichthyoxenus amurensis parasite from the Amur ide, Leuciscus waleckii, in Kherlen river, Mongolia. Амарын сугас (leuciscus waleckii) загасанд ichthyoxenus amurensis шимэгчийн үзүүлэх нөлөө Хураангуй: Загас нь хүнс тэжээлийн бас нэг баялаг бөгөөд олон зуун жилийн турш тэдгээрийг судалж, хамгаалан, ашиглаж ирсэн. Манай орны 3 ай савын гол, мөрөн, нуурт 2 анги, 8 баг, 13 овог, 44 төрөлд хамаарах 70 гаруй зүйл загас тархжээ. Хэрлэн голын дагуу хийгдсэн загасны паразитын судалгаагаар 3 зүйлийн загас илэрч байв. Судалгаанд хамрагдсан n=25 загасанд өвчний шинжилгээ хийхэд 8 тооны Амарын сугас (Leuciscus waleckii) загас Ichthyoxenus amurensis паразитаар халдварласан байв. Ichthyoxenus amurensis нь сугас загасны цээжний сэлүүрийн хоорондын болон хэвлийн арьсыг цоолон хэвлийн хөндийд нэвтэрдэг болох нь тогтоогдов. Ichthyoxenus amurensis зүйл шимэгчээр халдварласан загасны өсөлт удааширч, бие давжаарах, биеийн жин буурдаг болох нь батлагдав. Мөн Ichthyoxenus amurensis нь эм загасанд илүү шимэгчилж буйг судалгаагаар илрүүлэв. Хэрлэн голын сав газар дахь Амарын сугас загасанд Ichthyoxenus amurensis зүйл шимэгчилж буйг анх тутам илрүүлсэн болно. Түлхүүр үг: Ichthyoxenus amurensis, паразит, биеийн өсөлт, тарга авалт,

A recent study demonstrated lower incidence of fever and hepatosplenomegaly in HIV-positive VL patients than HIV-negative VL patients, indicating that acquired immunity have important roles in development of hepatosplenomegaly also in human VL. To explore the roles of acquired immunity in pathology of VL, pathological characterization of L. donovani infection in RAG2-/-mice was performed. One hundred million promastigotes were inoculated intraperitoneally to RAG2-/-mice as well as BALB/c mice, and organs were harvested from the animals at 2, 4, 8 and 12 weeks after infection and examined for parasite burdens and pathological changes. L. donovani infection induced splenomegaly in BALB/c mice; the length of the spleen after 12 weeks of infected mice was as twice as that of naive mice. In contrast, such the enlargement was not observed in RAG2-/-mice. Accumulations of mononuclear cells were found in the liver of infected BALB/c mice, whereas such the formations were less found in RAG2-/-mice. These results suggest that development of hepatosplenomegaly during experimental VL is dependent on the T cell or B cell rather than the presence of parasites.Mongolian Journal of Agricultural Sciences Vol.15(2) 2015; 43-50

Haemaphysalis longicornis is known as one of the most important ticks transmitting Babesia parasites in East Asian countries, including Babesia ovata and Babesia gibsoni, as well as Theileria parasites. H. longicornis is not the natural vector of Babesia bovis and Babesia bigemina. Vector ticks and transmitted parasites are thought to have established unique host–parasite interaction for their survival, meaning that vector ticks may have defensive molecules for the growth control of parasites in their bodies. However, the precise adaptation mechanism of tick-Babesia parasites is still unknown. Recently, cyclophilin A (CyPA) was reported to be important for the development of Babesia parasites in ticks. To reveal a part of their adaptation mechanism, the current study was conducted. An injection of B. bovis-infected RBCs into adult female H. longicornis ticks was found to upregulate the expression profiles of the gene and protein of CyPA in H. longicornis (HlCyPA). In addition, recombinant HlCyPA (rHlCyPA) purified from Escherichia coli exhibited significant inhibitory growth effects on B. bovis and B. bigemina cultivated in vitro, without any hemolytic effect on bovine RBCs at all concentrations used. In conclusion, our results suggest that HlCyPA might play an important role in the growth regulation of Babesia parasites in H. longicornis ticks, during natural acquisition from an infected host. Furthermore, rHlCyPA may be a potential alternative chemotherapeutic agent against babesiosis.

4E-BP, an eIF4E-binding protein, is well known as a cap-dependent translation inhibitor. Here, the 4E-BP homolog, Hl4E-BP, was isolated and identified from the hard tick Haemaphysalis longicornis. Hl4E-BP transcripts were ubiquitously expressed in the active stages, including the larvae, nymphs, and female adults, and the transcription levels were found to be higher in unfed than engorged ticks. In contrast, the expression levels of non-phosphorylated Hl4E-BP, which is a 13.4-kDa protein detected by the anti-recombinant Hl4E-BP antibody, were the highest in engorged ticks and significantly decreased progressively during the unfed starvation period of ticks. The functional role of Hl4E-BP as a metabolic brake was verified by histochemical observations on the lipid storage in midguts and fat bodies during the starvation period using ticks injected with dsHl4E-BP. The results indicate that Hl4E-BP is highly relevant to the lipid storage of ticks during the non-feeding starvation period. Our results suggest, for the first time, that Hl4E-BP may have a crucial role in the starvation resistance of ticks in an off-host condition via lipid metabolism control, although it was unclear whether Hl4E-BP might be involved in lipid synthesis regulation and/or lipid consumption inhibition.

Vitellogenin (Vg) synthesis, vitellogenesis, is an essential process for the development and reproduction of ticks. Our previous finding led to the hypothesis that target of rapamycin (TOR) pathway is important for vitellogenesis in the hard tick, Haemaphysalis longicornis. The TOR pathway controls cellular activity according to nutrient availability in eukaryotes. TOR, a member of the phosphatidylinositol 3-kinase family, is a central player in this pathway. Here, we present preliminary evidence that H. longicornis TOR (HlTOR) controls vitellogenesis via activation of S6 kinase (S6K) in the fat body. RNA interference (RNAi)-mediated gene silencing of HlTOR was undertaken to elucidate the involvement of HlTOR in the vitellogenesis of the tick. HlTOR-RNAi caused inhibition of S6K phosphorylation in the fat body. HlTOR-RNAi also altered not only the expression levels of GATA mRNA and protein but also the intracellular localisation of GATA in the fat body. The expression levels of Vg mRNA and protein in the fat body of HlTOR-RNAi ticks were significantly lower than those in control ticks. In the pre-ovipositional stage, the ovaries of control ticks had brown oocytes developing, but those of HlTOR-RNAi ticks were white and immature. The haemolymph colour indicated that the amount of Vg was lower in HlTOR-RNAi ticks than in the controls. Furthermore, rapamycin inhibited S6K phosphorylation and reduced the expression levels of Vg mRNA and protein in the fat bodies. Vg proteins were not detected in rapamycin-treated fat bodies in the presence of 20-hydroxyecdysone. These results suggest that HlTOR activity is critical for vitellogenesis stimulated by 20-hydroxyecdysone.

Ixodid ticks transmit various pathogens of deadly diseases to humans and animals. However, the specific molecule that functions in the recognition and control of pathogens inside ticks is not yet to be identified. Class B scavenger receptor CD36 (SRB) participates in internalization of apoptotic cells, certain bacterial and fungal pathogens, and modified low-density lipoproteins. Recently, we have reported on recombinant HlSRB, a 50-kDa protein with one hydrophobic SRB domain from the hard tick, Haemaphysalis longicornis. Here, we show that HlSRB plays vital roles in granulocyte-mediated phagocytosis to invading Escherichia coli and contributes to the first-line host defense against various pathogens. Data clearly revealed that granulocytes that up-regulated the expression of cell surface HlSRB are almost exclusively involved in hemocyte-mediated phagocytosis for E. coli in ticks, and post-transcriptional silencing of the HlSRB-specific gene ablated the granulocytes' ability to phagocytose E. coli and resulted in the mortality of ticks due to high bacteremia. This is the first report demonstrating that a scavenger receptor molecule contributes to hemocyte-mediated phagocytosis against exogenous pathogens, isolated and characterized from hematophagous arthropods.

RNA interference is an efficient method to silence gene and protein expressions. Here, the class B scavenger receptor CD36 (SRB) mediated the uptake of exogenous dsRNAs in the induction of the RNAi responses in ticks. Unfed female Haemaphysalis longicornis ticks were injected with a single or a combination of H. longicornis SRB (HlSRB) dsRNA, vitellogenin-1 (HlVg-1) dsRNA, and vitellogenin receptor (HlVgR) dsRNA. We found that specific and systemic silencing of the HlSRB, HlVg-1, and HlVgR genes was achieved in ticks injected with a single dsRNA of HlSRB, HlVg-1, and HlVgR. In ticks injected first with HlVg-1 or HlVgR dsRNA followed 96 hours later with HlSRB dsRNA (HlVg-1/HlSRB or HlVgR/HlSRB), gene silencing of HlSRB was achieved in addition to first knockdown in HlVg-1 or HlVgR, and prominent phenotypic changes were observed in engorgement, mortality, and hatchability, indicating that a systemic and specific double knockdown of target genes had been simultaneously attained in these ticks. However, in ticks injected with HlSRB dsRNA followed 96 hours later with HlVg-1 or HlVgR dsRNAs, silencing of HlSRB was achieved, but no subsequent knockdown in HlVgR or HlVg-1 was observed. The Westernblot and immunohistochemical examinations revealed that the endogenous HlSRB protein was fully abolished in midguts of ticks injected with HlSRB/HlVg-1 dsRNAs but HlVg-1 was normally expressed in midguts, suggesting that HlVg-1 dsRNA-mediated RNAi was fully inhibited by the first knockdown of HlSRB. Similarly, the abolished localization of HlSRB protein was recognized in ovaries of ticks injected with HlSRB/HlVgR, while normal localization of HlVgR was observed in ovaries, suggesting that the failure to knock-down HlVgR could be attributed to the first knockdown of HlSRB. In summary, we demonstrated for the first time that SRB may not only mediate the effective knock-down of gene expression by RNAi but also play essential roles for systemic RNAi of ticks.

Ticks grow rapidly during blood feeding, and their body weight may ultimately increase 100-fold more than that before feeding. The molecular mechanisms controlling growth during blood feeding in ticks remain largely unknown. The conserved insulin/PI3K/Akt signaling pathway regulates growth and metabolism in eukaryotes. Here, we show evidence for the involvement of Akt in growth during blood feeding in the parthenogenetic strain of the hard tick Haemaphysalis longicornis. We identified a homolog of the Ser/Thr kinase Akt (HlAkt) from the EST database of the H. longicornis embryo. HlAkt cDNA had a 1,590 bp ORF that encodes 529 amino acids with a predicted molecular weight of 60 kDa. HlAkt possesses a PH domain, a Ser/Thr kinase domain, a hydrophobic motif, and dual phosphorylation residues (Thr 338 and Ser 503) that are essential for kinase activation. Knockdown of HlAkt by RNA interference caused inhibition of blood feeding in female ticks. Histological observation demonstrated that HlAkt knockdown led to the arrest of growth in internal organs. HlAkt knockdown also affected the expressions of blood meal-induced genes that are essential for blood digestion, development, and reproduction in the female tick. These results strongly indicate that HlAkt is essential to complete the blood feeding process accompanied by the growth of internal organs in adult ticks. This is the first report of identification and characterization of Akt in Chelicerata, including ticks.