The PCR-based detection of Babesia gibsoni infection in dogs (German shepherds) reared in South Korea

German-shepherd dogs from the South Korean provinces of Gangwon and Gyunggi were used in a study exploring both the detection of Babesia gibsoni infection, by use of a PCR-based assay, and the nucleotide sequences of part of the P18 gene of the parasite. Nine (1.8%) of the 501 dogs checked were found PCR-positive. Although the prevalence of the infection appeared slightly higher in the male dogs than in the female (2% v. 1.7%), and in the dogs aged <3 years than in the older animals (2.7% v. 1.1%), these differences were not statistically significant. In addition, the prevalences of infection among the dogs from Gangwon (in the east of the country) and those from Gyunggi (in the west) appeared similar. All nine PCR-positive dogs were confirmed to have B. gibsoni infection by the microscopical examination of thin bloodsmears, which revealed that 0.1%-3% (mean=1.5%) of the dog's erythrocytes harboured the parasite. The nucleotide sequences of the P18 gene of B. gibsoni from each of the PCR-positive dogs were identical to each other and to the corresponding sequence - recorded under accession number AB053292 in the DNA Data Bank of Japan (DDBJ) - previously determined for isolates of B. gibsoni from Japanese dogs. Although the PCR products sequenced in the present study represent only a part of the P18 gene of B. gibsoni, it seems that South Korean and Japanese dogs carry a similar strain of B. gibsoni. These results may help to clarify the phylogenetic position of the B. gibsoni found in South Korea.     

Relationship between Helicobacter pylori hopQ genotype and clinical outcome in Asian and Western populations

Background and Aims:  Outer membrane proteins of Helicobacter pylori mediate important pathogen–host interactions such as colonization, adhesion and the inflammatory response. hopQ genotypes have been suggested to be associated with increased risk of peptic ulcer. The aim of this study was to test the relation of hopQ genotype to H. pylori -related disease and histological changes in Asian and Western countries.
Methods:  hopQ genotype, cagA status and vacA genotype of H. pylori isolated from patients from Asian and Western countries were determined and the results were compared with the clinical presentation and gastric histology.
Results:  Most Asian strains possessed virulent genotypes ( hopQ type I, vacA s1-m1 and cagA -positive). In Western countries, hopQ type I genotype was significantly linked with vacA s1 and m1 genotypes and cagA -positive status. Inflammatory cell infiltration and atrophy scores were significantly higher in patients with hopQ type I strains than those with type II in Western patients. However, the hopQ type I genotype was not associated with an increased risk for peptic ulcer or gastric cancer, and had no additive effects to vacA genotypes or cagA -positive status.
Conclusion:  The expression of multiple putative virulence factors in Asian strains likely explains the relatively high incidence of clinical outcomes including gastric cancer compared with other parts of the world. Although hopQ genotype did not improve the predictive value above other genotyping for development of H. pylori -related gastroduodenal diseases, the hopQ genotype might be able to add a useful virulence marker for gastroduodenal diseases.     

利用BgAMA-1抗原建立吉布森巴贝斯虫的ELISA诊断方法

本研究利用基因重组技术,构建并表达吉布森巴贝斯虫的重组顶膜抗原-1(BgAMA-1),将BgAMA-1作为诊断抗原,建立针对吉布森巴贝斯虫的ELISA诊断方法。结果显示,构建的重组表达质粒得以表达,建立的ELISA方法在血清学诊断中有较好的特异性。     

Asian traditional medicine (ATM) based on in vivo microcirculation evidence

Asian traditional medicine (ATM) (herbal medicine, acupuncture or moxibution) has gained some popularity among communities in Asia, but there are still few evidences to prove the effectiveness of such therapeutic measures. A symposium was held with aim at the effectiveness of Asian traditional therapies in views of in vivo microcirculation. This report is concerned with the symposium, including Asian activities for ATM.     

The First Example of the Rh Antigen Ew in Western Europe

Abstract. A fifth white family — the first in Western Europe — carrying E^w is reported, the antigen being traced through three generations.     

Molecular evidence of Babesia equi transmission in Haemaphysalis longicornis

We studied the tick, Haemaphysalis longicornis, to determine the possibility of both transovarial and transstadial transmission of Babesia equi. We also studied the usefulness of the needle injection method for pathogenic tick-transmitted organisms including Babesia parasites. Erythrocytes infected with B. equi were injected into the midgut of engorged adults or nymphs using a hypodermic needle passed through the integument. DNA of B. equi in ticks was detected using nested polymerase chain reaction (PCR). B. equi DNA was present in adults, eggs, and larvae, indicating that transovarial transmission occurred. B. equi DNA was present in adults that developed from infected nymphs, and the B. equi antigen was present in their salivary glands, indicating that transstadial transmission occurred. These findings suggest that H. longicornis may play a role in the transmission of B. equi.     

Identification of a novel gene encoding a secreted antigen 1 of Babesia gibsoni and evaluation of its use in serodiagnosis

Serum from a dog immunized with blood plasma from a B. gibsoni-infected dog, putatively containing secreted antigens, was used to screen a cDNA expression library. A novel gene encoding BgSA1 was identified from the isolated clones. The serum raised in mice immunized with the recombinant BgSA1 expressed in Escherichia coli could recognize a native parasite protein with a molecular mass of 59 kDa. Comparing with the previously established ELISA with recombinant P50 as antigen, the ELISA with recombinant BgSA1 as the antigen was more sensitive when they were used to detect field samples. Moreover, a sandwich ELISA with anti-BgSA1 antibodies could detect the circulating BgSA1 in a serial blood plasma from a dog experimentally infected with B. gibsoni. These results indicated that BgSA1 could be a useful target for the development of a diagnostic test for the detection of specific antibodies and circulating antigens.     

No evidence of deer mouse involvement in plague (Yersinia pestis) epizootics in prairie dogs

Plague, the disease caused by the bacterium Yersinia pestis, can have devastating impacts on black-tailed prairie dog (Cynomys ludovicianus) colonies. One suggested mechanism behind sporadic prairie dog die-offs involves an alternative mammal host, such as the deer mouse (Peromyscus maniculatus), which often inhabits prairie dog colonies. We examined the flea populations of deer mice to investigate the potential of flea-borne transmission of plague between deer mice and prairie dogs in northern Colorado, where plague is active in prairie dog colonies. Deer mice were predominantly infested with the flea Aetheca wagneri, and were rarely infested with prairie dog fleas, Oropsylla hirsuta. Likelihood of flea infestation increased with average monthly temperature, and flea loads were higher in reproductive animals. These results suggest that the deer mouse is an unlikely maintenance host of plague in this region.     

First evidence of transmission of Leishmania (Viannia) lainsoni in a Sub Andean region of Bolivia

Using ubiquitous primers which amplify the variable parts of kDNA minicircle of all Leishmania spp, we obtained for Leishmania (viannia) lainsoni a major band of 605 bp (band 1) shared with L. V. braziliensis and a minor 524 bp band (band 2) specific of L. V. lainsoni. The specificity of the two bands was examined through Southern blot hybridization of kDNA PCR obtained from reference strains belonging to L. braziliensis, L. mexicana, L. donovani complexes with L. V. lainsoni species. Band 1 was not specific of L. V. lainsoni since it hybridized with some isolates belonging to L. braziliensis complex. In contrast, band 2 was L. V. lainsoni specific. PCR-based detection followed by hybridization with the new L. V. lainsoni probe (Band 2) and L. V. braziliensis probe (564 bp), was assayed using sample from a pool of 25 females of Lutzomiya nuneztovari anglesi, blood, skin and liver samples of 18 mammals, spinal cords of four mammals and blood and cutaneous ulcers aspirates from 95 patents from Sub Andean region of La Paz, Bolivia. We observed a ositive hybridization of four patients lesions and the pool of L. nuneztovari anglesi with the L. V. lainsoni probe. It is the first time that L. V. lainsoni is observed in a cycle of transmission in Bolivia. PCR products of three patients lesions and the pool of L. nuneztovari anglesi were also hybridized with the specific probe of L. V. braziliensis suggesting mixed infection in this focus.     

Hepatitis C virus infection in Western Europe

Hepatitis C virus (HCV) infection is now recognised as the most common infection causing chronic liver disease in Europe. Approximately 3% of the world population has been infected with HCV, which represents about 170 million chronic carriers at risk of developing serious complications with more than 5 million in Europe alone. In the general population, the prevalence varies geographically from about 0.5% in northern countries to 2% in Mediterranean countries. Among newly detected HCV cases, 40-60% have normal ALT levels, 80% are viraemic, while about 70% of newly detected HCV carriers present histologic liver disease. More than 75% of the cases remain to be identified. The residual risk for transmitting HCV by blood products is at present 1/200 000 units distributed. Intravenous drug users are currently the main risk group. The prevalence rate is about 80% and the yearly incidence varies between 4 and 6%. In haemodialysis patients, the prevalence ranges from 10% to 30% and the incidence from 3% to 7%. The source of infection for the 30% of cases without identifiable risk factor remains to be clarified and appropriate well-controlled case-control studies on large samples are necessary. Further training and information campaigns remain desirable to improve knowledge and awareness among health care professionals.     

Vaccination of dogs against Babesia canis infection using parasite antigens from in vitro culture

Groups of five dogs were vaccinated with different Babesia canis vaccine formulations. It appeared that partial protection against challenge infection was obtained when using parasite antigens from in vitro culture in combination with saponin. Protection was evident as a decrease in parasitaemia after challenge and was associated with the presence of serum antibodies against Babesia parasites. In addition, parasite antigen derived from in vitro culture supernatant exhibited more protective activity than somatic parasite antigen, in that a less marked fall of haematocrit values was found after challenge infection. The fall of haematocrit value observed in the animals immunized with somatic parasite antigen was not different from that observed in the adjuvant control group.     

First Record of Mosquito-Borne Kyzylagach Virus in Central Europe

RNA of Kyzylagach virus (KYZV), a Sindbis-like mosquito-borne alphavirus from Western equine encephalitis virus complex, was detected in four pools (out of 221 pools examined), encompassing 10,784 female Culex modestus mosquitoes collected at a fishpond in south Moravia, Czech Republic, with a minimum infection rate of 0.04%. This alphavirus was never detected in Central Europe before.     

Short report: cloning of the Babesia gibsoni cytochrome B gene and isolation of three single nucleotide polymorphisms from parasites present after atovaquone treatment

We determined the nucleotide sequence of the Babesia gibsoni cytochrome b (cytb) gene. DNA was extracted from B. gibsoni isolated from Aomori Prefecture, Japan, and 1,288 basepairs of the cytb gene, including 1,071 basepairs of the open reading frame, were sequenced. The cytb gene of B. gibsoni obtained from three dogs that had been experimentally infected with B. gibsoni and treated with atovaquone was also sequenced. The B. gibsoni cytb gene obtained from all three atovaquone-treated dogs contained a single polymorphism resulting in an amino acid change in one of the putative ubiquinone-binding sites of Plasmodium falciparum. This polymorphism was homologous to mutations in other apicomplexan protozoa that exhibit resistance to atovaquone. Two other single polymorphisms were identified in parasites isolated from two of the dogs. These results indicate that single nucleotide polymorphisms in the sequence for mitochondrial cytb gene may be associated with decreased susceptibility of Babesia species to atovaquone.