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Background. In Thailand, Japanese encephalitis appears with a higher incidence of the Northern region (0.0/100,000 cases/year) as compared to the central southern region of the country (0.01/100,000/year). Also, all Japanese encephalitis virus (JEV) recently isolated strains in country belong to the genotype I and III.Methods. In order to evaluate the long-term efficacy of the newly developed live attenuated SA 14-14-2 Japanese encephalitis vaccine in Thailand, the immune response in children was studied after a second immunization campaign. Following 2012 immunization campaign, a second JEV immunization vaccine was delivered to children among four provinces of the Northern Region of Thailand. For each province, the recipients were identified accordingly to the two vaccine campaigns time including the children who received the second dose of vaccine: 1/ less than one year before the present study (N=30 by province); 2/ more than one year before the present study (N=70 by province). Altogether, a total of 400 children were enrolled in this project. After receiving the second vaccine dose, blood samples were collected and tested for JEV (Genotypes I and III) neutralizing antibodies following a standard procedure of LLC-MK2 infected cells.Results. All recipient presented neutralizing antibodies cross reacting against the prevalent JEV genotypes I (SM1 JEV strain) and genotype III (SA 14-14-2 and Beijing JEV strains). Most of the children from the first group had a seroconversion rate of 94.2% against homologous (i.e. GIII). Among them, an optimal rate of 100% seroconversion was find at the Phayao Province against both heterologous (GI) and homologous (GIII) JEV strains. Geometric Mean Titer (GMT) of neutralizing antibody against SM1, SA 14-14-2 JEV, and Beijing JEV strains were consistently high, respectively as 239±50, 323±201.2 and 177±36.7. For the second group, seroconversion rate was lower against SM1, SA 14-14-2 and Beijing JEV strains with a rate of 94.3, 90% and 86.8% with respectively a GMT of 154±32, 90±23 and 82±13.6.Conclusion. Although, the immunity appears to decrease within the period of 2.5 years, neutralizing antibody rate are consistent to protect against the JEV infection. Nevertheless, a long-term follow up is suitable to evaluate an eventual third dose opportunity.

Broiler is among Thai major industries. Red blood cell (RBC) count is one of the basic health screening protocols. The counting is often done manually. The mammalian blood analyzer cannot be used, because the shapes of mammalia and avian RBCs are different. RBC's color also varies among staining methods. Furthermore, RBCs can be grouped together. In this paper, we propose the automatic method for counting avian RBC. Otsu's multiple thresholding is used to extract the nucleus of RBC. The problems of the varying color of and the conglomerated RBC are tackled by applying iterative process. Only one parameter needs to be set manually. The experiment on five different staining types demonstrated that our proposed method provided less than 5% counting error for the staining that did not have the dark RBC's boundary.

Red blood cell counting is one of the most requested blood tests and can be effectively done by the dedicated equipment. In this paper, we proposed the two-stage circular Hough transform as the alternative automatic method for canine red blood cell counting. The possible blood cell area in the input slide image is first extracted. In the first stage, the Circular Hough transform with low sensitivity is applied to detect the non-overlapped cells. The detected cells are removed. The Circular Hough transform with high sensitivity is re-applied to the remaining region in order to detect the overlapped cells. The proposed method can be easily implemented into personal computers. The proposed algorithm was tested with many slides of canine red blood cell. It provided the counting with the average error of less than 5% with the best result of 1.439% error.

International audience; Background: Chikungunya virus (CHIKV) outbreak recurrences in Thailand are unpredictable and separated byunexplained and often long silent epidemiological periods that can last for several years. These silent periods could beexplained in part by the fact that infection with one CHIKV strain confers lasting natural immunity, even against otherCHIKV strains. In this study we evaluated the persistence of CHIKV-specific neutralizing antibodies in the population ofChumpae District, Khon Kaen Province, nineteen years after a CHIKV outbreak occurred in the same area in 1991.Findings: Overall 39% (44/111) of 111 former patients had neutralizing antibodies reacting against CHIKV ECSAstrain. Consistently high titers of neutralizing antibodies were found in 75% (33/44) of all positively-reacting sera,70% of which (23/33) were collected from individuals amongst the >60 years old age group. Although theprevalence found in Pong Haeng village (70%) was significantly higher than the prevalence detected in the NongThum village (14%), control study villages without known previous Chikungunya epidemics had a high Chikungunyaneutralizing antibody prevalence (65%).Conclusions: More than one-third of the pre-exposed population had persisting natural immunity that was morelikely boosted by recent and repetitive exposure to the emerging ECSA CHIKV in Thailand. Also, Chikungunyavirus appears to largely circulate in the country with a great variability appears between villages or area probablyassociated with the vector abundance and efficiency. Altogether these results show a potential for a lifelongimmunity against CHIKV. Given the rapid spread of the highly pathogenic ECSA strain in Southern Thailand, thedevelopment of CHIK vaccine is strongly recommended.

The potential of macaque Macaca nemestrina leonina in Thailand to be infected by endemic arboviruses was assessed. The prevalence of antibodies of three arboviruses actively circulating in Thailand was determined by Plaque Reduction Neutralization assay procedures using samples from captive colonies in Northern Thailand. Out of 38 macaques, 9 (24%) presented reacting antibodies against dengue virus, 5 (13%) against Japanese encephalitis virus, and 4 (10%) against Chikungunya virus. Our results indicate that the northern pig-tailed macaque in Thailand can be infected by these arboviruses, inferring therefore that their virus specific vectors have bitten them. Given that, northern pig-tailed macaque represents an abundant population, living in close range to human or in peridomestic setting, they could play a role as potential reservoir host for arboviruses circulating in Thailand.

Twenty-nine domestic piglets from pig farms located in three provinces of Thailand between 2003 and 2004 were used as sentinel animals for Japanese encephalitis virus (JEV) circulation. Piglets were used as sentinel to underline, on one hand, the role of domestic pigs as JEV amplifying host and, on another hand, to point out the interest of using sentinel animals for Japanese encephalitis surveillance. JEV activity was demonstrated through i/ antibody detection using a specific ELISA test for the identification of Immunoglobulins of class M and G, ii/ virus isolation on cell culture, after experimental mosquito inoculation for virus amplification. Almost 100% and 83% of the piglets, respectively, had specific IgG and IgM JEV antibodies and 35% yielded a virus isolate. Piglets of the growing farm industry act as virus amplifier increasing the risk of transmission for the human community. Conclusively, since piglets JEV infection appears early in life and is generally clinically unnoticed, it represents an exceptional sentinel model for human health threats, which has to be considered by health authorities.