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4 & Journal of Mahanakorn Veterinary Medicine Editorial Advisory Board President of Mahanakorn University of Technology Dean of Faculty of Veterinary Medicine Editor Jamlong Mitchaothai D.V.M., M.Sc., Ph.D. Assistant Editor Rachakris Lertpatarakomol B.Sc., M.Sc. Somchai Chanket D.V.M. Editorial Board Thanongsak Mamom D.V.M., Ph.D. Sumrarn Bunnajirakul D.V.M., Dr.med.vet. Araya Suepkhampet D.V.M., Ph.D. Jetsada Rungpupradit D.V.M., Dr.med.vet. Phinidda Cha-umphol D.V.M., M.Sc. Sukanya Phalitakul D.V.M., M.Sc. Chulabha Sonklein B.Sc., M.Sc. Thuchadaporn Chaikhun D.V.M., M.Sc. Jitra Sanisuriwong D.V.M. Waradee Buddhakosai D.V.M. Chainarong Phumratanaprapin D.V.M., M.Sc. Tassanee Triratapiwan B.Sc., M.Sc. Warissa Sangkaew D.V.M. Titaree Laoharatchatathanin D.V.M. Supparat Wannasilp D.V.M. 4

5 ก ก ก ก ก ก ก ก ก ก ก ก 34/ ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก 5

6 & 4 1 ก 2552 ISSN : ก Ryan O Handley, Andrew Thompson, ก ก ก ก -ก ก กก ก ก ก ก : กก ก ก ก ก ก ก ก ก ก ก : ก Structures, Localizations and Functions of Taste Receptors Kazumi Taniguchi and Kazuyuki Taniguchi ก ก ก What s your cytological diagnosis? ก

7 Journal of Mahanakorn Veterinary Medicine Volume 4 No. 1 January June 2009 ISSN : Contents Research Cultivation of Neospora caninum in Vero Cell Jitbanjong Wiengcharoen, Ryan O Handley, Andrew Thompson, Parntep Rattanakorn and Yaowalark Sukthana Seroprevalence of Foot-and-Mouth Disease in Cattle and Buffaloes at the Kanchanaburi Animal Quarantine Station During Sorayut Seekhaow and Apiradee Intarapuk Preliminary Study: Nutritional Value of By Product from Pangasius bocourti Processing Chanathip Thammakarn, Nahatai Vijitrothai and Chunya Kongrith Meat Quality Examination of Pork and Beef in Local Fresh Market and Supermarket in Muang District and Hun Hin District of Phetchaburi Province Bhutharit Vittayaphattananurak Raksasiri and Rachakris Lertpatarakomol Case Report Treatment of Chronic Trypanosomiasis in Dog: A Case Report Eakkasit Barameechaithanun, Pimsuda Suwannasaeng, Nittaya Boonbal, Sudarat Pattanee and Sompong Hoisang Review Article Structures, Localizations and Functions of Taste Receptors Kazumi Taniguchi and Kazuyuki Taniguchi Brucellosis in Goats Sommai Kaopiew and Komkrich Pimpukdee Q & A What s your diagnosis? Titaree Laoharatchatathanin and Thanate Anusaksathien What s your cytological diagnosis? Hassadin Boonsriroj and Thanongsak Mamom

8 & ก ก 4 1 ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก Kazumi Taniguchi ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก 2552 ก / ก ก ก ก ก ก ก ก ก ก ก 8

9 ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก (Research Papers) ก ก ก 1.2. (Case Report) ก ก ก ก 1.3. (Articles) ก ก ก ก ก ก ก 2.2. ก 3 ก Browallia new 16 ก 4 ก ก ก ก ก ก ก (Electronic file) (PC formatted CD) ก Microsoft Word (Window 98 ก ) ก ก ก ก 3. ก 3.1. (Title) ก 9

10 & 3.2. (Author and Co-authors) ก ก ก ก ก (Corresponding author) ก 3.3. (Abstract) ก ก ก ก 3.4. (Keywords) ก ก ก 5 ( ) ก 3.5. (Introduction) ก ก (Literature review) ก ก 3.6. ก ก (Materials and Methods) ก ก ก ก ก ก 3.7. ก (Results) ก ก ก ก ก ก ก (Figures) ก ก ก ก ก ก 3.8. (Discussion) ก ก ก ก ก ก ก 3.9. (Conclusion) ก ก ก ก ก ก 3.10.ก ก ก (Acknowledgement) ก ก ก (References) 10

11 4. ก - ก กก ก (Cited by) - ก ก (2549) ก (, 2549), ( ก, 2549) - ก ก Tomato and Danny (2006), Taylor et al. (2006) ก (Tomato and Danny, 2006), (Taylor et al., 2006) - ก (Personal comm.) ก - ก ก ก ก ก ( ก ก ) ( ) ก ก ก ก ก ก ก ก. ก. 18(3) : Excellenta, S.A., Excellente, S.B. and Excellento, S.C Researcher performances for publishing in a journal. J Excellence Eaa. 78 : ก ก ก ( ก ) ก ก ก กก ก ก ก. ก ก. ก. 125 Excella, H.A Expertization by reading. 4 th edition. Angthong. A.B.C. publishing. Philad. 430 p. 11

12 & ก 140. ก ก ก ก 2. ก ก ก 12

13 ก 1, #, Ryan O Handley 2, Andrew Thompson 2, ก 3 ก 4 (Neospora caninum) ก ก ก ก 20 ก ก ก ก ก ก ก ก ก ก ก ก ก กก ก ก ก ก ก ก ก ก ก ก ก (neosporosis) ก ก ก ก กก ก ก ก ก ก ก K9WA tachyzoite (vero cell) CO 2 incubator :,, tachyzoite, 1 2 Division of Veterinary and Biomedical Sciences, Murdoch University, Western Australia 3 4 # / 13

14 & 14 (Neospora caninum) (obligate intracellular parasite) ก ก ก ก ก ก (encephalomyelitis) (Bjerkas et al., 1984) ก ก ก (Toxoplasma gondii) ก ก ก ก ก ก Dubey ก (Dubey et al., 1988) กก ก ก (neosporosis) ก ก (abortion) ก ก ก ก ก ก ก ก ก ก (stillbirth), ก ก ก ก (congenital neosporosis), ก (reduce milk production) ก ก (increase culling) ก (Anderson et al., 2000; Gottstein et al., 2000; Dubey et al., 2006; Dubey et al., 2007) ก ก ก ก ก ก ก ก ก ก ก Kyaw et al. (2004) seropositive 30 (5.5%) ก 549 ก competitive ELISA (celisa) seropositive 1 (1.2%) ก 82, Suteeraparp et al. (1999) seropositive 6% ก 904 ก 11 ก Indirect Immunofluorescence Antibody Test (IFAT) Kashiwazaki et al., (2001) seropositive ก % 83 ก IFAT ก 3 tachyzoite, tissue cyst oocyst ( 1) tachyzoite tissue cyst ก ก ก ก ก ก tachyzoite ก 6 x 2 tissue cyst ก ก (round or oval) 107 ก CNS ก 4 bradyzoite 7-8 x 2 ก bradyzoite ก ก ก tachyzoite (Buxton et al., 2002; Dubey, 2003) oocyst ก ก coyote ก ก ก oocyst

15 ก 11.7 x 11.3 (McAllister et al., 1998; Lindsay et al., 1999; Bergeron et al., 2001; Buxton et al., 2002) 3 (infective stage) ก ก cyst bradyzoite ก ก oocyst ก ก ก tachyzoite ก ก ก (transplacental transmission) ก transplacental transmission ก ก ก 1 Neospora caninum (Dubey, 2003). ก ก N.caninum strain K9WA tachyzoite vero cell ก ก The Animal Health Laboratories, Agriculture Department, Western Australia ก humidified incubator 37 C, 5%CO 2 ก Inverted microscope ก 2 vero cell ก infected N.caninum ก tachyzoite ก monolayer ก ก ( 2) cell scraper monolayer ก ก flask medium tachyzoite N.caninum ก hemocytometer ก inoculate N.caninum 10 5 tachyzoites vero cell monolayer ก flask humidified incubator 37 C, 5%CO 2 ( 3) ก subculture ก vero cell ก 15

16 & ก DMEM/HIGH w/o L-Glutamine (Hyclone) ก 2 Growth culture medium fetal calf serum 10% Maintenance culture medium fetal calf serum 2% ก ก vero cell กก inoculate ก ก inoculate vero cell ก N.caninum strain K9WA tachyzoite ก ก ก ก ก ก 3-4 passages ก ก ก subculture ก tissue culture N.caninum ก Inverted microscope tachyzoite ก vero tachyzoite vero cell ก ( ก ) 16

17 3 tissue culture flask N.caninum DMEM humidified incubator ก ก ก ก ก ก กก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก กก ก ก ก ก ก N.caninum 3 tachyzoite, bradyzoite oocyst ก ก ก ก ก ก ก กก ก ก ก ก ก tachyzoite ก ก bradyzoite ก ก ก ก ก กก ก tachyzoite T.gondii ก ก N.caninum ก bradyzoite ก tachyzoite ก ก (Dubey et al., 1998) ก bradyzoite ก กก tachyzoite ก ก tachyzoite bradyzoite ก phase contrast Giemsa, PAS, silver stains ก (immunostaining) ก ก ก TEM (Dubey et al., 1998) 17

18 & ก ก bradyzoite N.caninum ก ก mice, gerbil ก ก ก ก bradyzoite ก ก ก ก ก N.caninum (Gottstein et al., 2000) ก oocyst ก ก ก ก ก coyote ก oocyst ก ก ก ก กก oocyst oocyst ก ก oocyst ก ก ก (McAllister et al., 1998; Lindsay et al., 1999; Dubey, 2003; Dubey et al., 2007) N.caninum tachyzoite กก ก ก (Wiengcharoen et al., 2007) ก ก ก ก ก กก ก ก ก Anderson M.L., Andrianarivo A.G., Conrad P.A Neosporosis in cattle. Anim Reprod Sci.60-61: Bergeron N., Fecteau G., Villeneuve A., Girard C., Pare J Failure of dogs to shed oocysts after being fed bovine fetuses naturally infected by Neospora caninum. Vet Parasitol. 97: Bjerkas I., Mohn S.F., Presthus J Unidentified cyst-forming sporozoon causing encephalomyelitis and myositis in dogs. Z. Parasitenkd. 70: Buxton D., McAllister M.M., Dubey J.P The comparative pathogenesis of neosporosis. Trends Parasitol.18: Dubey J.P Review of Neospora caninum and neosporosis in animals. Korean. J Parasitol. 41:1-16. Dubey J.P., Carpenter J.L., Speer C.A., Topper M.J., Uggla A Newly recognized fatal protozoan disease of dogs. J Am Vet Med Assoc. 192:

19 Dubey J.P., Lindsay D.S., Speer C.A Structures of Toxoplasma gondii tachyzoites, bradyzoites, and sporozoites and biology and development of tissue cysts. Clin Microbiol Rev. 11: Dubey J.P., Buxton D., Wouda W., Pathogenesis of bovine neosporosis. J Comp Path. 134: Dubey J.P., Schares G., Ortega-Mora L.-M., Epidemiology and control of neosporosis and Neospora caninum. Clin Microbiol Rev. 20: Gottstein H.B., Conraths F.J., De Meerschman F., Ellis J.T., Innes E.A., McAllister M.M, Ortega-Mora L.-M., Tenter A.M., Trees A.J., Uggla A., Williams D.J.L., Wouda W A European perspective on Neospora caninum. Int J Parasitol. 30: Kashiwazaki Y., Pholpark S., Charoenchai A., Polsar C., Teeverapanya S., Pholpark M Postnatal neosporosis in dairy cattle in northeast Thailand. Vet Parasitol. 94: Kyaw T., Virakul P., Muangyai M., Suwimonteerabutr J Neospora caninum seroprevalence in dairy cattle in central Thailand. Vet Parasitol. 121: Lindsay D.S., Dubey J.P., Duncan R.B Confirmation that the dog is a definitive host for Neospora caninum. Vet Parasitol. 82: McAllister M.M., Dubey J.P., Lindsay D.S., Jolley W.R., Wills R.A., McGuire A.M Dogs are definitive hosts of Neospora caninum. Int J Parasitol. 28: Suteeraparp P., Pholpark S., Pholpark M., Charoenchai A., Chompoochan T., Yamane I., Kashiwazaki Y Seroprevalence of antibodies to Neospora caninum and associated abortion in dairy cattle from central Thailand. Vet Parasitol. 86: Wiengcharoen J., O Handley R., Armstrong T., Best W., Sukthana Y., Thompson R.C.A Novel drug compounds against Neospora caninum and Toxoplasma gondii in vitro. Southeast Asian J Trop Med Public Health. 38 (suppl 1):

20 & Cultivation of Neospora caninum in vero cell Jitbanjong Wiengcharoen 1,#, Ryan O Handley 2, Andrew Thompson 2, Parntep Rattanakorn 3 and Yaowalark Sukthana 4 Abstract Neospora caninum was first reported as the causative agent of encephalomyelitis in dog since Nowaday, it was recognized as a protozoa which can cause an important reproductive disease, neosporosis, in cattle throughout the world. It can also cause disease in dogs, goats, sheep, horses and deer. There were many reports concerned with neosporosis in cattle but its life cycle, biology and pathogenesis was not yet completely understood until now. The cultivation of N.caninum in cell culture would be very useful because the parasite can be used as the source of antigen and DNA for the diagnostic purpose. Moreover, the lived parasite can be used to infect the animals in the experiment to get more understanding about its biology and pathogenesis. This study is the first report in Thailand that success to culture the tachyzoite stage of N.caninum strain K9WA in vero cell. 20 Keywords : Neospora caninum, neosporosis, tachyzoite, vero cell 1 Faculty of Veterinary Medicine, Mahanakorn University of Technology 2 Division of Veterinary and Biomedical Sciences, Murdoch University, Western Australia 3 Faculty of Veterinary Sciences, Mahidol University 4 Department of Protozoology, Faculty of Tropical Medicine, Mahidol University # Corresponding Author /

21 ก ก ก ก ก กก ก ก 2,# ก ก ก ก ก ก ก ก กก ก ก ก ก ก ก ก ก ก ก ก ก ก กก ก ก ก ก ELISA Non-structure 3 ABC, 3B protein (NSP) ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก (p < 0.05) ก ก ( 8.6) ก ก ก ก ( 12.4) (p < 0.05) : ก, ก, non-structural protein, ELISA, ก 1 ก กก ก ก 2 #, 21

22 & ก ก ก ก ก ก (Alexanderson and Mowat, 2005) ก ก ก ก ก ก ก Foot-and-Mouth disease virus (FMDV), Genus Aphthovirus, Family Piconaviviridae (King, 2000) ก ก ก ก ก ก ก ก ก ก ก ก ก กก ก FMDV ก ก 7 ก O, A, C, Asia1, SAT1, SAT2 SAT3 ก 3 ก A, O Asia1 ( ก, 2549;, 2541) (Klein et al., 2008; Perez et al., 2005; Tosh et al., 2002; Kesy, 2002) ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ( ก, 2549;, 2541) กก ก (2549) ก ก ก ก Virus Infection Associated Antigen (VIA) test ก (2549) ก ก 2548 ก NSP 4 ก ก ก ก ก กก ก ก ก ก ก ก ก ก ก ก ก ก กก ก ก (OIE, 2002) ก ก ก ก ก ก ก กก ก ABC 3B ELISA ก ก ก ก ก (Shen et al., 1999) ก ก ก ก ก ก ก กก ก ก ก ก 22

23 ก ก ก ก ก ก ก ก กก ก ก ,076 ก ก ก ก ก FMDV ก ก - ก CHEKIT FMD-3ABC UBI FMDV NONSTRUCTURAL PROTEIN ELISA ก CHEKIT FMD-3ABC bo-ov (Switzerland) indirect ELISA 3ABC Nonstructural protein FMDV ก ก ก Positive Negative control Value% = ODsample - ODneg x 100 ODpos - ODneg ก Value% <20% ก FMDV = 20 30% ก FMDV > 20% ก FMDV ก 3B nonstructural protein FMDV indirect ELISA UBI FMDV NONSTRUCTURAL PROTEIN ELISA (CATTLE) ก ก ก cut-off point OD ก ก ก NS (3ABC, 3B-ELISA) test FMDV ก ก ก ก ก ก Epi info version odd ratio 95% ก ก ก ก ก ก กก ก 8,076 ก , ,136 ก NS test ก NS test 23

24 & ก ก NS test ก NS test 2.97 ก ก NS test ก NS test ก ก NS test 2550 (P < 0.05) ก ก ก ก NS test ( 8.6) ก ก ก NS test ( 12.41) (P < 0.05) ( 1) 1 ก NS test ก ก NS test ก -ก ( ) ก NS test ( ) ก NS test ก ก ก a b c d a,b c,d ก ก (p < 0.05) ก 1 ก ก ก FMDV 2549 ก ก FMDV ก ก ( 32.77) ก ก FMDV ก ก ก 2550 ก ก 2549 ก FMDV ก ก 2550 ก FMDV ก ก ก ก FMDV ก ก ก ก FMDV ก กก ก 2550 ก ก ก ก ก

25 1 เปอร เซ นต ม.ค. ก.พ. ม.ค. เม.ย. พ.ค. ม.ย. ก.ค. เด อน ส.ค. ก.ย. ต.ค. พ.ย. ธ.ค. ก NS test พ.ศ.2549 พ.ศ เปอร เซ นต ม.ค.-49 ม.ค.-49 พ.ค.-49 ก.ค.-49 ก.ย.-49 พ.ย.-49 ม.ค.-50 ม.ค.-50 พ.ค.-50 ก.ค.-50 ก.ย.-50 พ.ย.-50 % โคต ดเช อ % กระบ อต ดเช อ ก ก NS test ก ก ก ก กก ก 2549 ก NSP 14 ก ก NS FMDV กก 10 ก ก ( 2530) ก 2549 (7 12 ) ก NSP FMDV กก 10 ก ก ก ก ก ก ก ก ก ก ก ก ก (2549) ก ก ก 2548 ก NSP 4 ก ก ( กก 10) 25

26 & ก ก (p < 0.05) ก ก ก ก ก ก ก (Green et al., 2006) ก ก ก ก ก ก ก ก ก ก ก ก ก (Gibbens et al., 2001) ก ก ก ก กก ก ก ก (Salt, 1993) ก ก ก ก (Vasloo et al., 1996) ก ก ก ก (Esophageo-pharyngeal) ก (Salt, 1993; Anderson et al., 1979) ก ก ก กก ก Vasloo et al. (1996) ก ก ก ก FMDV ก ก ก ก ก ก ก ก ก FMDV ก Nonstructural protein (NSP) ก ก ก FMDV ก ก กก ก (Sorensen et al., 1998; Malirat et al., 1998; De Diego et al., 1997; Bergmenn et al., 1993; Berger et al., 1990) ก ก ก (false positive) ก ก ก (Bergmenn et al., 1993) กก ก ก ก ก ก ก NS protein ก ก ก ก ก ก ก Liquid phase blocking (LP) ELISA ก FMDV (, 2537;, 2541; ก, 2549) ก ก ก ก ก ก ก กก กก กก (Hedger, 1970) ก ก ก NS protein ก ก ก ก ก ก Structural protein ก ก NS protein ก กก กก ก กก 3ABC NS protein ก ก ก FMDV ก ก ก ก ก ก FMDV 3ABC ELISA

27 (sensitivity) (specificity) ก ก 100 ก Virus neutralization test (VNT) (Bronsvoort et al., 2004) ก ก ก ก 3ABC protein ก ก ก ก ก ก (Lubroth and Brown, 1995; Lubroth et al., 1996) ก ก NS protein ก ก ก 3B NS protein ก ก 94 ก (Shen et al., 1999) ก NSP FMDV ELISA ก ก ก ก ก ก ก ก กก ก ก ก ก ก Non structural protein ก ก ก ก ก (Office International des Epizooties; OIE) ก ก (Food and Agriculture Organization; FAO) ก ก ก ก ก ก กก กก ก ก ก ก ก ก กก ก ก NSP ก ก ก กก กก ก NSP ก ก ก ก กก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก กก ก ก ก ก ก ก ก ก ก ก ก NSP ก ก ก ก ก ก ก ก กก ก NSP ก 27

28 & FMDV ก ก ก FMDV Liquid phase blocking ELISA ก ก กก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก FMDV..... ก ก ก ก ก ก ก -ก. ก. ก ก ก ก 6. ก. ก ก ก ก ก ก ก ก. ก ก 1. ก. ก ก ก ก ก ก ก [Online]. Available: ก ก ก. ก 1. ก ก. ก. ก ก ก ก Alexandersen, S. and Mowat, N Foot and mouth disease: host range and pathogenesis. Curr Top Microbiol Immunol. 288: Anderson, C.E., Docherty, W.J. Anderson, J. and Paling, R The pathogenesis of foot-and-mouth disease in the African buffalo (Syncerus caffer) and the role of this species in the epidemiology of the disease in Kenya. J Com Pathol. 89:

29 Berger, H.G., Straub O.C. Ahl, R., Tesar, M. and Marquardt, O Identification of Foot-and-mouth disease virus replication in vaccinated cattle by antibodies to nonstructural virus proteins. Vaccine. 8: Bergmenn, I.E., de Mello, P.A., Neizert, E., Beck, E. and Gomes, I Diagnosis of persistent aphthovirus infection and its differentiation from vaccination response in cattle by use of enzyme-linked immunoeletrotransfer blot analysis with bioengineered nonstructural viral antigens. Am J Vet Res. 54: Bronsvoort, B.M.C., Sorensen, K.J., Anderson, J., Corteyn, A., Tanya, V.N., Kitching, R.P. and Morgan, K.L A comparison of two 3 ABC ELISA s in a cattle population with endemic, multiple serotype foot-and-mouth disease. J Clin Microbiol. 42: De Diego, M., Brocchi, E., Mackay D. and De Simone, F The non-structural polyprotein 3ABC of foot-and-mouth disease virus as a diagnostic antigen in ELISA to differentiate infected from vaccinated cattle. Arch Virol. 142: Gibbens, J.C., Sharpe, C.E., Wilesmith, J.W., Mansley, L.M., Michalapoulou, E., Ryan, J.B.M. and Hudson, M Descriptive epidemiology of the 2001 foot-andmouth disease epidemic in Great Britain: the first five months. Vet. Res. 149: Green, D.M., Kiss, I.Z. and Kao, R.R Modeling the initial spread of foot-and-mouth disease through animal movements. Proc. R. Soc. 273: Hedger, R.S Observations of the carrier state and related antibody titres during an outbreak of foot-and-mouth disease. J Hyg Camb. 68: Kesy, A Global situation of foot-and-mouth disease (FMD)-a short review. Pol J Vet Sci. 5: King, A.M.Q Picornaviridae (Seventh report of the International Committee for the Taxonomy of Viruses). Virus Taxonomy: Classification and Nomenclature of Viruses. Van Regenmortel, M.H.V., Fauquet, C.M. and Bishop, D.H.L., eds. Academic Press. pp Klein, J., Hussain, M., Ahmad, M., Afzal, M. and alexandersen, S Epidemiology of foot-and-mouth disease in Landhi dairy colony, Pakistan, the world largest buffalo colony. Virol J. 5: 53. Lubroth, J. and Brown, F Identification of native foot-and-mouth disease virus nonstructural protein 2C as a serological to differentiate infected from vaccinated livestock. Res Vet Sci. 59:

30 & Lubroth, J. Grubman, M.J., Burrage, T.G., Newman, J.F.E. and Brown, F Absence of protein 2C from clarified foot-and-mouth disease virus vaccines provides the basis for distinguishing convalescent from vaccinated animals. Vaccine. 14: Malirat, V., Neitzert, E., Bergmenn, I.E., Maradei, E. and Beck, E Detection of cattle exposed to foot-and-mouth disease virus by means of an indirect ELISE test using bioengineered nonstructural polyprotein 3ABC. Vet Q. 20: S24-S26. Office International des Epizooties Report of the second OIE/FAO-APHCA workshop on WTO s Sanitary and Phyto-Sanitary (SPS) agreement. OIE representation for Asia and the Pacific, Tokyo, Japan and animal production and health commission for Asia and the Pacific. 293 p. Perez, A.M., Thurmond, M.C., Grant, P.W. and Carpenter, T.E Use of the scan statistic on disaggregated province-based data: foot-and-mouth disease in Iran. Prev Vet Med. 71: Salt, J.S The carrier state in foot-and-mouth disease-an immunological review. Br Vet J. 149: Shen, F., Chen, P.D. Walfield, A.M. Ye, J. and House, J Differentiation of convalescent animals from those vaccinated against foot-and-mouth disease by a peptide ELISA. Vaccine. 17: Sorensen, K.J., Hansen, C.M., Madsen, E.S. and Madsen, K.G Blocking ELISAs using the FNDV nonstructural proteins 3D, 3 AB and 3ABC produced in the Baculovirus expression system. Vet Q. 20: S17-S20. Tosh, C., Hemadri, D. and Sanyal, A Evidence of recombination in the capsid-coding region of type A foot-and-mouth disease virus. J Gen Virol. 83: Vasloo, W., Bastos, A.D., Kirkbride E., Esterhuyen, J.J., Janse van Rensburg, D., Bengis, R.G., Keet, D.W. and Thomson, G.R Persistent infection of African buffalo (Syncerus caffer) with SAT-type foot-and-mouth disease viruses: rate of fixation of mutations antigenic change and interspecies transmission. J Gen Virol. 77:

31 Seroprevalence of Foot-and-Mouth disease in Cattle and Buffaloes at the Kanchanaburi Animal Quarantine Station during Sorayut Seekhaow 1, Apiradee Intarapuk 2, # Abstract Foot and mouth disease (FMD) is a high contagious viral disease which endemic in Thailand. This study focused on the Kanchanaburi Animal Quarantine Station, located on Thai-Myanmar border, in which a large number of animals had immigrated from Myanmar. Serum of cattle and buffaloes were collected from apparently healthy each month from 2006 to Samples were detected antibodies against FMDV by the Non-structural 3 ABC, 3B protein (NSP) ELISA assay that has greatly advanced sero-diagnosis/surveillance as the test detects exposure to live virus for any the seven serotypes of FMDV, even in vaccinated populations. Thoughtfully the study, the FMDV infectious prevalence was 10.29%, as well as 14.49% were found in 2006 and 3.67% were found in The prevalence in 2006 was significantly higher than 2007 (p < 0.05). Cattle and buffaloes were found infections in 8.6% and 12.4%, respectively. The statistical analysis showed significantly difference of infectious rates between cattle and buffalo (p < 0.05). Keywords : Foot and mouth disease, prevalence, non-structural protein, ELISA, Kanchanaburi 1 Kanchanaburi International Animal Quarantine Station, Department of Livestock Development 2 Faculty of Veterinary Medicine, Mahanakorn University of Technology # Corresponding author, 31

32 & ก ก : กก ก 1,# 1 1 ก ก (Pangasius bocourti) กก (Filet) 7 3 ก ก 1 6 ก Proximate analysis (Moisture) (Dry matter) (Ash) (Ether extract) (Crude protein) ก ก ก (Crude fiber) (Calcium) (Phosphorus) ก (Gross energy) Automatic Bomb Calorimeter กก ก 49.35% 50.65% 4.13% 31.88% 13.41% 3,687 kcal/kg 1.47 % % 7.58 % % 46.93% 0.34 % 6.12 % 0.62 % 6,770 kcal/kg ก ก กก ก ก ก ก 32 : 1 ก ก ก ก ก #

33 (Pangasius bocourti) 1 ก ก ก ก (, 2538) ก ก ก ก (Filet) ก ก ก (Pangasius hypophthalmus) ก ก ก (Moller, 2007) ก ก ก ก ก ก ก ก ก ก ก ก กก ก 2 ก ก ก ก ก ก ก ก ก ก กก ก ก ก ก ก ก กก ก ก ก ก ก ก ก ก ก ก ก ก ก กก ก ก ก กก ( ก ก ก ) 7 3 ก ก ก ก ก ก 60 (2-3 ) ก ก ก ก ก ก ก ก ก 33

34 ournal of Mahanakorn Veterinar Medicine Vol. 4 No. 1 anuar - une 2009 ÃÙ» Õè 1 áê»åòà¼òð Õ è ºÑ ÁÒ ÒกกÃÐªÑ กè͹กÒêÓáËÅÐ ÃÙ» Õè 2 áê àèéªô¹é Êèǹ»ÅÒà¼ÒÐ Õàè ËÅ Í ÒกกÒêÓáËÅЪԹé Êèǹ๠Íé (Filet) กÒÃÇÔà ÃÒÐËì ³ Ø èò Ò âહð Í µñçíâèò 1. ÓกÒÃÇÔ à ÃÒÐËì µ Ñ Ç ÍÂè Ò Õ è à µãõ  ÁäÇé â ÂÇÔ Õ proximate analysis â  ÓกÒÃÇÔ à ÃÒÐËì ྠèíëò ÇÒÁª é¹ (Moisture) Çѵ ØáËé (Dry matter) à éò (Ash) ä Áѹ (Ether extract) â»ãµõ¹ (Crude protein) Ñ é µñ Ç ÍÂè Ò ã¹êàò¾ê áåðµñ Ç ÍÂè Ò ã¹êàò¾áëé ºÒ Êè Ç ¹ áåð ÓกÒÃÇÔ à ÃÒÐËì ËÒà Íè ãâ (Crude fiber) á Åà«ÕÂÁáÅÐ ÍÊ ÍÃÑÊÊÓËÃѺµÑÇÍÂèÒ ã¹êàò¾áëé ºÒ Êèǹ 34

35 2. ก (Gross energy) ก Automatic Bomb Calorimeter (LECO AC-350 Automatic Bomb Calorimeter) ก ก ก ก ก กก % 50.65% 4.13 % % % 3,687 kcal/kg 1.47 % % 7.58 % % % 0.34 % 6.12 % 0.62 % 6,770 kcal/kg 1 ก กก ก (31.88% % ) ก ก ก (13.41%) ก (46.93%) ก ก ก ก ก ก ก กก (4.13%) ก (7.58%) ก ก ก 6.12% 0.62% ก ก ก ( ก ก ) 10 1 ก ก (3,687 kcal/kg) (6,770 kcal/kg ) ก ก ก ก ก ก ก Adipose tissue ก (2550) ก Adipose tissue 35

36 & 1 ก กก 36

37 ก ก กก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก กก ก ก ก ก ก ก ก ก ก ก กก ก ก ก ก Men et al. (2007) ก ก ก ก (Backfat) ก ก ก ก ก ก ก ก ก (fish oil) ก (2006) ก ก กก 10% Docosahexaenoic acid (DHA) 2% Total Omega 3 กก 3% ก ก กก ก ก ก กก ก ก ก ก ก ก ก ก ก ก ก ก กก ก ก ก ก ก ก ก.. ก ก ก ก ก ก ก ก 37

38 & ก : ก 3. : ก ก ก ก ก ก ก ก ก. ก ก 22/2538. ก. ก. ก ก ก ก ก. [Online] Available : 8/04/2009 Men L. T., S. Yamasaki, H. H. Chi, H. T. Loan and R. Takada Effects of Catfish (Pangasius hypophthalmus) or Coconut (Cocos nucifera) Oil, and Water Spinach (Ipomoea aquatica) in Diets on Growth/Cost Performances and Carcass Traits of Finishing Pigs. JARQ 41 (2) : Moller A., Seafood Processing Local Sources, Global Markets. Organisation for Economic Co operation and Development. 36 p. 38

39 Preliminary Study: Nutritional Value of By Product from Pangasius bocourti Processing Chanathip Thammakarn 1,#, Nahatai Vijitrotai 1 and Chunya Kongrith 1 Abstract Nutritional value of by product from Pangasius bocourti processing was studied. The by product compose of fish meat together with fish head and spine after filet separated out. Seven samples were randomly collected and three fishes in each. One of these samples was collected as fresh sample and the six other samples were kept as partially dried. Proximate analysis method was used for determining the nutritional value moisture, dry matter, ash, ether extract, crude protein. Crude fiber, calcium and phosphorus were measured in partially dried samples only. Gross energy was determined by Automatic Bomb Calorimeter. The results reveal that the fresh sample has 49.35% moisture, 50.65% dry matter, 4.13% ash, 31.88% ether extract and 13.41% crude protein. The gross energy of the fresh sample is 3,687 kcal/kg. For the partially dried samples, the averages of nutritional value are 1.47% moisture, 98.53% dry matter, 7.58% ash, 46.38% ether extract, 46.93% crude protein, 0.34 of crude fiber, 6.12% calcium, 0.62% phosphorus and gross energy density equals 6,770 kcal/kg. The nutritional value disclosed that the by product from Pangasius bocourti processing is possible to apply for animal feed. As the results of its nutritional value, the by - product can be used for energy and protein source and may serve as a raw material ground head and fish spine, because of high content of calcium and phosphorus. Therefore, it is possible to use the partially dried by product as a raw material or for developing products for human. Keywords : Nutritional value, By product, Pangasius bocourti 1 Division of Animal Production Technology and Fisheries, Faculty of Agricultural Technology, King Mongkut s Institute of Technology Ladkrabang # Corresponding author : 39

40 & ก ก ก ก ก 1,# ก ก 2 ก ก ก ก Salmonella spp. ก ก (CFU/g) ก ก ก (P<0.01) ก 5.92x10 5 CFU/g 2.16x10 5 CFU/g ก ก (P<0.01) ก 5.01x10 5 CFU/g 1.68x10 5 CFU/g (CFU/g) ก ก (P<0.05) ก 5.44x10 5 CFU/g 1.40x10 5 CFU/g ก ก ก (P>0.05) ก 4.47x10 5 CFU/g 4.05x10 5 CFU/g ก Salmonella spp. ก ก ก ก Salmonella spp. ก ก : ก 40 1 ก ก ก 2 #

41 ก ก ก ก ก กก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก Escherichia coli Salmonella spp. ก ก ก ก ก กก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก Escherichia coli Salmonella spp. ก 2 ก ก ก ก ก ก ก ก ก ก ก ก ก 0-4 0, 3 5 ก (Completely Randomized Design, CRD) ก ก 4 ก ก ก 1 ก 2 ก 3 ก ก 4 ก 41

42 & ก 1. (Total Plate Count Total Viable Count) 25 ก buffer peptone water 225 ก 1-2 ก pour plate (, 2547;, 2539) 2. ก Salmonella Shigella (SS) ก ก (Qluashi, 1978; Jay, 2000; FDA, 1992) Triple sugar ironagar (TSI) K/A H 2 S Lysine decarboxylase agar (LIA) H 2 S ก Salmonella spp. (, 2547) ก ก ก ก Duncan s New Multiple Range Test ก SAS version 6.12 ก 1. ก 1.1 ก กก ก ก ก ก ก 0 3 ก ก ก ก (P<0.01) ก ก ก 0 ก 5.92x10 5, 5.01x10 5,1.68x x10 5 CFU/g ก ก ก 3 ก 6.48x10 5, 6.08x10 5, 3.44x x10 5 ก ก ก 5 ก ก ก ก (P<0.01) ก 9.92x10 5, 7.64x10 5, 6.68x x10 5 CFU/g ก ก ก ก ก 1 42

43 1 (x 10 5 CFU/g) ก ก ก ก ก ก ( ) a 6.48 a 7.64 b 1.68 b 3.44 b 4.92 c ก 2.16 b 3.36 b 9.92 a ก 5.01 a 6.08 a 6.68 b abc ก ก ก ก ก (P<0.01) 1.2 กก ก ก ก ก 0 ก ก ก (P<0.05) ก 4.05x10 5, 5.44x10 5, 4.47x x10 5 ก ก ก 3 ก ก ก (P<0.01) ก 8.64x10 5, 4.84x10 5, 4.28x x10 5 ก ก ก 5 ก ก ก (P<0.01) ก 8.84x10 5, 4.92x10 5, 4.44x x10 5 ก ก ก ก ก 2 2 (x 10 5 CFU/g) ก ก ก ก ก ( ) d 4.84 b 4.92 b 4.47 d 8.64 a 8.84 a ก 1.40 e 4.28 b 4.44 b ก 4.05 d 2.60 c 4.20 b abc ก ก ก ก ก (P<0.01) de ก ก ก ก ก (P<0.05) 43

44 & 2. ก 2.1 ก ก กก ก ก ก 3 SS agar, TSI LIA ก SS agar ก ก TSI ก ก H 2 S LIA ก H 2 S ( 3) ก SS agar ก ก TSI ก ก H 2 S ก ก LIA ก H 2 S ( 3) ก ก SS agar ก ก TSI ก LIA ก H 2 S ( 3) ก ก SS agar ก ก TSI ก ก H 2 S ก ก LIA ก H 2 S ( 3) 3 ก ก K = (slant); A = ก (butt); Gas = ก ก ; H 2 S = ก 44

45 2.2 ก ก ก ก SS agar ก ก TSI ก ก H 2 S ก ก LIA ก H 2 S ( 4) 4 ก ก 1 2 SS TSI LIA SS TSI LIA K/A + H 2 S K/A + H 2 S gas,h 2 S gas,h 2 S A + H 2 S K/A + H 2 S gas,h 2 S gas,h 2 S ก K/A + H 2 S K/A + H 2 S gas gas,h 2 S ก K/A gas,h 2 S + H 2 S K/A gas,h 2 S - K = (slant); A = ก (butt); Gas = ก ก ; H 2 S = ก ก ก ก ก ก ก 1.68x x10 5 CFU/g ก 1.40x x10 5 CFU/g ก ก (2547) ก ก 2.5x x10 7 CFU/g ก ก ก ก ก ก กก ก 25 ก ( ก, 2547) ก ก ก ก ก ก (, 2532) ก 1-10 ก (Doyle et al., 1997) ก ก ก ก 45

46 & ก ก (Jernklinchan et al., 1994; Bangtrakulnonth et al., 1994) ก 86 ก ก ก ก ก (, 2539) ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก (, 2545) ก ก ก ก ก ก ก ก ก ก ก ( ก, 2544) ก ก ก กก ก ก ก ก ก ก ก ก ก ก (, 2547) กก ก ก ก ก ก ก ก ก ก ก ก ก 7 ก ก ก ก 10 ก ก ก (, 2545) กก ก Smulders and Woolthuis (1985) กก ก ก ก ก ก ก ก (2545) ก ก ก ก ก ก ก ก ก ก 2% (v/v) ก ก ก ก ก ก ก ก ก ก 5 ก ก (, 2545) Ziauddin et al., (1993) ก ก ก 2% (v/v) ก Staphylococcus aureus, Salmonella Newport, Streptococcus faecali, Bacillus cereus, Pseudomonas fragai E. coli ก ก 24 46

47 ก ก ก ก ก ก ก กก ก ก ก. ก ก ก 34 (1) : ก ก ก ก ก ก ก ก, ก ก ก ก ก ก Salmonella, ก ก ก, ก , , ก. ก, ก. ก ก ก ก Standard Conventional Modified Semisolid Rappaport Vassiliadis. 26 (2) : ก ก ก. ก ก Bangtrakulnonth, A., Boonmar, S., Marnrim, N., Leungyoslouechakul, S., Sutanthavibul, J. and Kusum, M., Study of pig Salmonellosis in Thailand.p.220. In Proc. 13 th IPVS Congress. Doyle, M.P., Beuchat, L.R. and Montville. T.J., Food microbiology fundamentals and frontiers. ASM Press. Washington D.C FDA Bacterial Analatical Manual. Food and Drug Administration/Association of official Analytical Chemists, Washington, D.C. Jernklinchan, J., Koowatananukul, K. and Saitanu, K., Occurrence of Salmonella in raw broilers and their products in Thailand. J. Food. Prot. 57 : Quashi, M Manual for the Laboratory Diagnosis of bacterial food poisoning and the assesssment of the sanitary quality of food. The Southeast Asian Medical Information Center/ International Medical Foundation of Japan, Japan. Smulder, F.J.M. and Woodthuis, C.H.J Immediate and delayed microbiological effect of lactic acid decontamination of calf carcasses influence an conventionally boned versus hot-bonong and vacumm packaged cuts. J. Food Prot. 48 (10), Ziauddin, K.S., Rao, D.N. and AmLA, B.L In vitro study on the effect of lactic acid and sodium chloride on spoilage and pathogenic bacteria of meat. J. Food Sci Technol. 30(3),

48 & Meat Quality Examination of Pork and Beef in Local Fresh Market and Supermarket in Muang District and Hun Hin District of Phetchaburi Province. Bhutharit Vittayaphattananurak Raksasiri 1 and Rachakris Lertpatarakomol 2 Abstract Hygiene factor examination of pork and beef meat was studied in Local fresh Market and Supermarket in Muang District and Hun Hin District of Phetchaburi Province. Total microbial count by using total plate count and isolation of Sallmonella spp. was carried on in laboratory. The results from total count of microbial contamination (CFU/g) in pork meat from local fresh market in Muang district Phetchaburi Province was highly significance than supermarket meat in Muang district Phetchaburi (5.92 x 10 5 CFU/g and 2.16 x 10 5 CFU/g, respectively) (P<0.01). Incontrast, supermarket in Hua Hin district was highly significance than Hun Hin district (5.01 x 10 5 CFU/g and 1.68 x 10 5 CFU/g, respectively) (P<0.01). Total microbial contamination (CFU/g) in Beef Meat from local fresh market in Muang district Phetchaburi was highly significance than Supermarket in Muang district Phetchaburi (5.44x10 5 CFU/g and 1.40x10 5 CFU/g, respectively) (P<0.01). Hua Hin district Market and supermarket in Hua Hin district is not significantly difference. (4.47x10 5 CFU/g and 4.05 x 10 5 CFU/g, respectively) (P<0.01). Isolation of Sallmonella spp. in pork and beef from local fresh market and supermarket in Muang district Phetchaburi Province and Hun Hin district found that all tested samples was contaminated by Sallmonella spp. Keywords : Salmonella spp., pork, beef meat, microbial 1 Faculty of Animal Science and Agricultural Technology, Silpakorn University Phetchaburi IT Campus. 2 Faculty of Veterinary Medicine, Mahanakorn University of Technology. # Corresponding author. 48

49 - 1 ก 2 1 ก ก 2 ก ก 5 6 ก ก ก ก 1 ก ก ก ก ก ก ก ก ก กก ก ก ก ก ก ก ก ก ก ก ก ก ก ก 1.5 ก ก (Meanace reflex) ก (Pupillary light reflex) ก ก (Fluorescein) ก ก ก 1 2 ก ก ก ( ก 79) 49

50 & What s your cytological diagnosis? 1 ก 1, 2 1 ก ก ก ก ก ก ก กก ก 1.5. ก ก ก ก ก ก ก ก ก 3 ก ก ก ก ก (Impression smears) Modified wright stain (Diff-Quick ) ก 1-2 ( ก ) ก ก ( ก 81)

51 ก ก ก 1,# ก 37.6 ก ก ก 1 ก ก ก ก ก ก ก ก ก ก ก ก ก Trypanosoma evansi ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก : 1 ก ก # : 51

52 & Trypanosomiasis ก ก ก ก (kinetoplast) ก DNA ก (flagellum) ก ก undulating membrane ก ก ก ก ก ก ก ก ก Trypomastigote ก ก ก ก 1 ก parasitemia ก 1-3 ก ก กก Trypanosoma evansi ก ก ก ก ก ก ก ก (aqueous humor) ก ก ก ก ก ก ก ก ก (nictitating membrane) (conjunctiva) ก ก ก ก ก ก ก ก ก ก 3 ก ก coma ก ก ก ก ก กก ก (Brun et al., 1998;, 2540) ก ก ก Diminazene aceturate 3-7 ก ก ก ก 2 ก ก (Ghaffar, 2000)

53 ÊѵÇá¾ ÂìÁËÒ¹ ÃÊÒû è 4 ºÑº è 1 Á ÃÒ Á - ÁÔ ¹Ø Ò¹ 2552»ÃÐÇѵÊÔ µñ Çì»Çè  ÊØ¹Ñ ª èí»õàµíãì ÍÒÂØ 2»Õ ÃÖè ¾Ñ¹ ØìÃçÍ äçàåíãì à¾è¼ùé ¹éÓ˹Ñก 37.6 กÔâÅกÃÑÁ à éòãñºกòã ÃÑ ก ÉÒ Õ è â à ¾ÂÒºÒÅÊÑ µ Çì ³ÐÊÑ µ Çá¾ ÂÈÒʵÃì ÁËÒÇÔ ÂÒÅÑ Â Í¹áกè ¹ àá è Í ÇÑ ¹ Õ è 26 กÃก Ò Á ¾.È éçâíòกòãºçá¹éóºãôàç³ Ò Ëͺ ÁÕä é àâ èíàá ÍกÁÕÊÕàËÅ Í «Õ à»ç¹áò»ãðáò³ 1 à Í ¹ ÊØ ¹ Ñ à Âä é Ã Ñ º กÒÃÃÑ ก ÉÒÁÒ Òก Õ è Í è ¹ áåé Ç ËÅÒ ÃÑ é «Ö è ÍÒกÒÃกç Â Ñ äáè Õ Ö é ¹ àåõ é  ã¹à µâã èòêñµçìáåðáõ»ãðçñµàô ÂกԹ๠Íé Ôº กÒõÃÇ ÃèÒ กÒ ÍØ ³ ËÀÙ Á Ô Ã è Ò กÒ 104 Í ÈÒ Òàùäεì ÁÕ Í ÒกÒÃ«Ö Á ÁÕ Ê Õ à  è Í àá Í กà»ç ¹ ÊÕ à ËÅ Í «Õ ËÒÂã Ëͺ ÁÕÍѵÃÒกÒÃËÒÂã 57 ÃÑé µèí¹ò Õ ÍѵÃÒกÒÃàµé¹ Í ËÑÇã 82 ÃÑé µèí¹ò Õ ºÇÁ¹éÓºÃÔàdz Ò ÁÕÀÒÇÐáËé ¹éÓÃéÍÂÅÐ 5 ÅÓªèÍ éí ¾ºÇèÒ ÁÕกÒà ÂÒ ¹Ò Í µñº กÒÃÇÔ¹ Ô ÑÂáÂกâà âã ¾ÂÒ Ôã¹àÅ Í (Blood parasite) âã ÕËè ¹Ù (Leptospirosis) âã µñºíñกàêº (Hepatitis) âã äµçòâ (Renal failure) ÃÙ» Õè 2 ઠÍé ÃÔ»¾Òâ¹â«Á ÕÍè ÂÙãè ¹กÃÐáÊàÅ Í 53

54 & 1 ก ก 54 Items Results Results Reference Values* Day 0 Day 19 Hematology CBC PCV (%) Hb (g%) RBC count (x10 6 /mm 3 ) WBC count (x10 3 /mm 3 ) Platelet estimation Adequate RBC morphology Polychromasia, Anisocytosis Differential WBC Band neutrophil (cells/µl) Neutrophils(cell/µl) 89% (6,327) 49% (4,488) 2,750-12,850 Lymphocyte(cell/µl) 6% (426) 36% (3,297) 430-5,800 Monocyte(cell/µl) 3% (213) 4% (366) 50-1,400 Eosinophil(cell/µl) 2% (412) 10% (916) 100 1,250 Blood parasite examination Routine exam. Trypanosome evansi not found Clinical chemistry Creatinine (mg/dl) ALT (GPT) (U/L) 1, Blood sugar (mg%) Leptospira Ab Negative : Kahn ( 2005) ก ก กก 1 ก ก ก (Regenerative

55 anemia) ก ก ก ก ก ก ก ก ก ก ก ก polychromasia anisocytosis ก (ALT) ก ก ก ก ก ก ก ก Trypanosoma evansi ก Leptospira spp. ก ก ก ก ก ก ก ก ก D5-½S Dipyrone 25 ก ก ก ก cefazolin 25 ก ก ก ก ก Trypanosoma evansi ก ก ก ก ก ก Silymarin ก 1 2 Cephalexin ก 1 2 Folic-C ก ก ก Dipyrone 25 ก ก ก ก ก ก ก Dipyrone 25 ก ก ก ก Trypanosoma evansi Diminazene aceturate 5 ก ก ก ก ก ก (D5-½S) ก 10 ก (kidmin) 10 (intralipid 10%) 5 ก Trypanosoma evansi 2 ก 8 ก ก ก ก ก Diminazene aceturate 5 ก ก ก ก Silymarin ก ก 10 ก ก ก ก ก ก Trypanosoma evansi ALT ก ก ก ก ก ก ก ก ก ก ก 55

56 & ก normocytic hypochromic anemia ก ก ก กก ก ก ก ก ก ก regenerative anemia ก ก ก ก polychromasia anisocytosis ก ก ก ก polychromasia anisocytosis regenerative anemia ก ก ก reticulocyte count ก regenerative non-regenerative anemia ก กก ก ก ก ก กก ก ก (lymphopenia) ก ก ก ก ก ก ก ( ก, 2545; Meyer and Harvey, 2004) ALT ก ก ก ก ก ก ก ก ก ก ก ก ก ก (, 2548) ALT ก ก ก ก Trypanosoma evansi ก ก ก ก Leptospira spp. ก ก ก ก Trypanosomiasis ก ก ก ก กก Trypanosoma evansi ก ก ก Variant Surface Glycoprotein (VSG) ก ก ก ก (Holmes, 1980; Hajduk, 1984) ก ก ก (anemia) ก ก ก ก ก ก ก ก กก ก ก immunoglobulin ก ก กก complement 56

57 ก ก ก ก ก erythrophagocytosis ก ก (Disseminated intravascular coagulation, DIC) ก ก ก haemolytic anemia ก ก ก mononuclear phagocytic system ก ก ก ก ก antigen-antibody reaction (Murrey, 1978) กก ก 2-3 C 2-3 ก ก ก ก 20 ก ก x 90 x ( ก - ) / 0.25 ก ก 30 ก ก 22 ก ก (, 2544) ก ก ก ก ก ก parasiticemia ก ก 1-3 ก ก ก ก ก pyruvate, acetate ก ก ก ก metabolism ก (, 2540) ก ก ก ก ก cephalexin ก cephalosporin 1 betalactamase ก ก ก ก ก Diminazene aceturate (Berenil ) ก ก Trypanosomiasis ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก (Donald, 2002) ก ก ก ก ก Silymarin antioxidant ก ก ก ก ก ก fatty degeneration change ALT ก ก ก ก (, 2548) กก 57

58 & ก ก ก (kidmin) (intralipid 10%) ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก (, 2540) ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก ก 58

59 ก ก ก. ก ก ก ก ก ก - ก ก. ก ก. ก. 250 ก ก ก ก ก. ก ก Brun, R., Hecker, H. and Lun, Z.R Trypanosoma evansi and T. equiperdum: distribution, biology, treatment and phylogenetic relationship (a review). Parasitology. 79: Donald, C.P Veterinary Drug Handbook. 4 th ed. Pharma. Veterinary Publishing, Minnessota. 993 p. Ghaffar, M. Parasitology Blood and tissue protozoa imaging [internet]. University of South Carolina School of Medicine; 2000 [cited 2006 May]. Available from: Hajduk, S Antigenic variation during the developmental cycle of Trypanosoma brucei. J Protozoa Tropi Anim. 31(1): Holmes, P.H Vaccination against trypanosomes. Symposia of the British Soceity for Parasitology p. Kahn, C.M. and Line, S The Merck Veterinary Manual 9 th ed. Merck&Co., Inc. Washington. 824 p. Meyer, D.J. and Harvey, J.W Veterinary Laboratory Medicine: Interpretation and Diagnosis. 3 rd ed. Saunders. Missouri. 351 p. Murrey, M Anemia of bovine African trypanosomiasis: An overview. In: Pathogenecity of Trypanosomes. Proceeding of a workshop held at Nairobi, Kenya November, p. 59

60 & Treatment of Trypanosomiasis in Dog Eakkasit Barameechaithanun 1,#, Pimsuda Suwannasaeng 1, Nittaya Boonbal 1, Sudarat Pattanee 1 and Sompong Hoisang 1 Abstract A two-years-old male Rottweiler 37.6 kg. body weight was brought to Khonkaen University Veterinary Teaching Hospital with the complaint of fever, edema and panting. He was treated by a previous veterinarian for 1 month without significant improvement. The physical examination showed 7% dehydration, mucous membranes are pale and jaundice, edema at mandible area and palpably enlarged liver. Hematological and Biochemistry were unremarkable except for anemia and increased alanineaminotransferase (ALT). Trypanosoma evansi was detected on a blood smear. Based on clinical and laboratory findings, the dog was put on dipyrone, diminazene aceturate, silymarin, cephalexin, folic-c, fluid therapy and given a blood transfusion. His clinical symptoms improved and Trypanosoma evansi was not detected in a blood smear after treatment. Keywords: Trypanosome, Anemia, Jaundice 1 Khonkaen University Veterinary Teaching Hospital, Khonkaen University, Khon Kaen, # Corresponding author: 60

61 Structures, Localizations and Functions of Taste Receptors Kazumi Taniguchi 1,# and Kazuyuki Taniguchi 2 1 Laboratory of Veterinary Anatomy, School of Veterinary Medicine, Kitasato University, Higashi, Towada, Aomori , Japan 2 Laboratory of Veterinary Anatomy, Faculty of Agriculture, Iwate University, Ueda, Morioka, Iwate , Japan Correspondence author: Kazumi Taniguchi, DVM, Ph.D. Phone: ext. 432 Fax: Purpose In this article, we describe the recent progress of taste receptor research in the scientific community. As of late, research on taste receptors has made remarkable progress. In the present review, we focus on taste receptors, paying special attention to receptors for sweetness, bitterness and umami. The Five Elements of Taste Up to now, five elements of taste have been reported: sweetness, bitterness, sourness, saltiness and umami. They are independent of each other. In addition to these five elements, other factors, such as the texture and temperature of food as well as fat, contribute to the final taste, thus resulting in a very complicated combination of mitigating factors. Sweetness: The sweetness receptors sense sugars and carbohydrates. Once we eat sugars and/or carbohydrates, our bodies utilize them as sources of energy. Sweetness receptors must have developed to sense molecules carrying information regarding sources of energy for our bodies. As a result, humans crave sweet food, since energy is essential to maintaining life. Everyone, even newborn babies, loves sweet foods. Bitterness: The bitterness receptors sense molecules present in foods that contain alkaloids or that are burnt black. Bitterness can alert us that a food is potentially toxic, so we can stop ingestion before being poisoned. Many individuals do like some slightly bitter 61

62 & foods, such as dark chocolates, coffee, or beer. Adults learn, as they grow up, that certain bitter foods are not toxic. However, small children usually do not like bitter foods. Sourness: The sourness receptors sense concentrations of hydrogen ions in food, alerting us that the food may be rotten or unripe. For example, un-ripened fruits are sour, while ripened fruits are sweet. The result is that humans, as in the case of bitter foods, resist consuming sour foods. Furthermore, as in the case of bitter foods, we learn that some sour foods, such as yogurt, can be safe. Saltiness: The saltiness receptors sense concentrations of sodium ions. Mammalian bodies require sodium ions to maintain ion balance, which is essential for their lives. Excess sodium intake, however, can prove harmful. Umami: The umami, or savory receptors sense amino acids. Meat and fish contain a lot of umami. Some vegetables, such as tomatoes, have a great deal of umami as well. Humans do not taste proteins, because the molecular sizes of proteins are too big to bind to receptors; instead, humans sense amino acids. Beef, for example, obtained from a freshly slaughtered cow, is not that tasty. However, if the beef is aged at a low temperature, enabling proteins to break down and, as a result, producing more amino acids, the taste of the beef improves greatly. This is because animals and humans can sense larger amount of amino acids. Amino acids, and therefore proteins, are essential as the building blocks of life, so humans have evolved to crave umami. History of Umami Research The word umami, meaning savory taste, comes from the Japanese language. A Japanese researcher first extracted a substance containing umami, monosodium glutamate (MSG), from seaweed. The researchers who declared for the first time that umami must be one of the taste elements, were also Japanese. Since then, Japanese researchers have conducted a large portion of the ongoing umami research. At the early stages of umami research, western researchers doubted the existence of umami. However, umami has now been established worldwide as one of the five taste elements. Umami Receptors and Inosine Monophosphate The umami receptors sense not only amino acids, but also inosine monophosphates (IMP) in the cytoplasm. IMP is generated by the degradation of adenosine tri-phosphates (ATP). ATP breaks down, becoming first adenosine di- 62

63 phosphate (ADP), then guanosine monophosphate (GMP), and finally IMP. The higher the amount of IMP in a food, the better it tastes. Another interesting aspect of IMP is the fact that the combination of IMP and amino acids clearly has synergistic effects on umami reception. Food containing only amino acids or only IMP can be relatively tasty. Taste improves greatly, however, when food contains both of these substances. Nerve responses of the Chorda tympani, connects taste buds to the brain, were investigated after an administration of either (1) only IMP; (2) a single type of amino acid, such as glutamic acid, serine, alanine, glycine, asparagine or lysine; or (3) combinations of IMP and a type of amino acid (Nelson et al., 2002) The results demonstrated that the level of nerve responses was raised when IMP was administered at the same time as the amino acid. Also investigated were the effects of IMP on other substances, such as sodium chloride and sucrose. The results revealed that IMP had no significant effect on the nerve responses. Humans generally combine IMP and amino acids in food preparation. For example, in traditional Japanese cooking, we combine kelp soup and dried bonito soup to make a more savory soup. This is the most common and basic method of Japanese cooking. Researchers have reported that kelp is rich in glutamic acids, while dried bonito is rich in IMP (Brand et al., 1989; Filer et al., 1979; Yamaguchi et al., 1991) As a result, this traditional combination in Japanese cooking is, scientifically, a very reasonable way to improve taste. People utilize this idea when we deal with meat as well. The concentration of IMP is greater in aged meat than in fresh meat. As for the reasons why we prefer aged meat, we could suggest at least three: (1) softer texture, (2) a higher concentration of amino acids, and (3) a higher concentration of IMP. The softer texture is brought by the degradation of cells and tissues caused by proteinases, enzymes which localize in lysosomes in cells alive. After the cells die, the lysosomal enzymes are released into the cytoplasm and break down the cells/tissues. The higher concentration of amino acids is brought on by the degradation of proteins. The higher concentration of IMP is brought on by the degradation of ATP. The speed of the ATP degradation after slaughtering differs greatly, depending on animal species. The amount of IMP reaches a maximum eight to ten hours after death in yellowtail tuna (fish); three days after slaughter in pigs, and seven to ten days after slaughter in cows. 63

64 & Taste and Evolution More than 25 different types of bitterness receptors have been discovered, but only three have been found for sweetness. This suggests that the ability to sense various types of bitterness might have once been essential for survival. If an animal or a species lacked the ability to sense a potentially toxic bitterness, it risked death or extinction. Only those who succeeded in sensing danger could have survived. As for saltiness, animals, including humans, are very sensitive to slight differences of salt concentrations in food. This sensitivity may be related to the evolution of vertebrates. Vertebrates evolved from ancient ocean fish, then went ashore to become animals that depended on fresh water. Salt concentrations in water must have been a matter of life or death for early vertebrates. This ability to detect water salinity may retain importance, even in today s mammals. 64 Taste Receptors Seven-transmembrane proteins and ion channels: There are two types of receptors, these for bitterness, sweetness and umami, composed of seventransmembrane proteins coupled with GTP binding-proteins, and those for saltiness and sourness, composed of ion channels. An alpha subunit of G-protein coupled with bitter receptors, alpha-gustducin, was discovered by McLaughlin, et al. (1992) as shown in figure 1. Taste receptors for sweetness, bitterness and umami are quite similar to those for smells. Taste receptors are quite similar to olfactory receptors in their structures. There are two types of olfactory receptors: (1) those on the olfactory epithelium lining the nasal cavity; and (2) those on the sensory epithelium of the vomeronasal organ. The former are for common smells, and the latter for pheromones. Both types of receptors are seventransmembrane proteins coupled with G-proteins, as are taste receptors. In both cases, information on taste or olfaction is transferred to the brain, processed there, and recognized as taste or smell. T1Rs and T2Rs: Researchers have categorized taste receptors into two families: T1Rs (T1R family) and T2Rs (T2R family). The T means Taste, and the R means Receptors. T1Rs are sweetness and umami receptors, while T2Rs are bitterness receptors (Chandrashekar et al., 2000; Kitagawa et al., 2001; Max et al., 2001; Nelson et al., 2001) Max et al. (2001) and Margolskee, R.F. (2002) suggested that T1Rs may take the form of homo- or hetero-dimers (Margolskee, 2002; Max et al., 2001) This means that the molecule is composed of two of the same or different units: the same units for the

65 homodimer and different units for heterodimer. Nelson et al. (2002) proved that a heterodimer, composed of T1R2 and T1R3, functions as a sweetness receptor while a heterodimer, composed of T1R1 and T1R3, functions as an umami receptor. Various methods have been utilized to prove the roles of taste receptors, such as heterologous expression and neurophysiological investigations. One method is to make taste genes express in cells which normally do not express them, such as human embryonic kidney cells, culture them, and examined them for changes of calcium ion concentrations after the administration of various types of taste substances. Another method is to measure the level of nerve responses using electro-physiological techniques after the administration of various taste substances on the tongue. Hetero-dimers The exact sequences of T1Rs are quite similar among animal species, yet slight differences still exist. Artificial sweeteners: Humans enjoy artificial sweeteners, such as aspartame. Actually, the market for artificial sweeteners is quite large because of weight-gain concerns. As described above, the sweet receptor is a hetero-dimer of T1R2 and T1R3. It is interesting to note, however, that mice do not sense the aspartame, although they are quite sensitive to natural sweeteners such as sucrose as well as other types of sweeteners, such as saccharin. When murine T1R2 is replaced by human T1R2, mice become able to sense the aspartame, proving that the site responsible for the perception of aspartame is located in the sequence of T1R2 which are different between mice and humans (Nelson et al., 2002) Thinking about species differences, we can imagine that other animals, but not humans, might have been enjoying new taste which humans have never even dreamed of. Cats: Animals belonging to the feline family, such as cats, lions, and tigers, lack the ability to sense sweetness. Dogs, on the contrary, crave sweet food. This phenomenon had intrigued researchers over the years. Recently, Lee et al. (2005) proved that a portion of feline T1R2 is different from that of animals that sense sweetness (Li et al., 2005) Localization of T1Rs and T2Rs Various types of T1Rs can co-localize in a single taste cell. Various types of T2Rs can similarly co-localize in a single taste cell. T1Rs and T2Rs, however, can never colocalize within a single cell. This means that while there are cells responsible for sensing 65

66 & various types of sweetness, and the cells responsible for sensing various types of bitterness, but there are no cells sensing both sweetness and bitterness at the same time. In our experience of cooking, we know that taste elements affect each other. For example, when Japanese people cook oshiruko, sweetened beans, a traditional Japanese sweet, chefs add small amount of salt into a pan when they cook beans with a large amount of sugar. The small amount of salt enhances the sweetness of the oshiruko. This enhancement is not taken place in a single taste cell, because the cells with sweetness receptors and the cells with saltiness receptors are different cells than each other. Interestingly, the taste cells with T1Rs and the cells with T2Rs can co-exist in a single taste bud, enabling the possibility that the mutual affection of different tastes might be taken place in taste buds. Alternatively, it may take place in the brain when the brain processes the taste information. Sensitivity to Taste Substances and Localization of the Receptors: Taste papillae: Taste receptors are localized on the apical cellular surface of taste cells in taste buds, which are localized in taste papillae. The taste papillae are widely distributed on the tongue and oral cavity. There are five types of papillae: fungiform, circumvallate and foliate papillae on the tongue, papillae on the soft palate of the oral cavity, and papillae on the epiglottis. Millar et al. (1986) counted the numbers of taste buds on human tongue tips and reported that it averaged 116 buds/cm 2 (Miller, 1986) Solitary some buds were reported even in the trachea (Tizzano et al., 2006) Different sensitivity to taste in different portions of the tongue: Kieshow (1894) s pioneer work revealed that the sensitivities for various taste elements differ among portions of the tongue (Kugino et al., 1997) Since then, a lot of research has been conducted to clarify which part of the tongue/oral cavity is responsible for which type of taste. The tip of the tongue is most sensitive to sweetness and saltiness, the lateral parts of the tongue are most sensitive to sourness, and the posterior portions of the tongue, close to the throat, are most sensitive to bitterness. The recent progress of taste receptor research has cast a new light on them. Localization of T1Rs in papillae: The distribution and densities of taste receptors are different depending on their location on the tongue and oral cavity (Nelson et al., 2001) T1R1: T1R1s are abundantly distributed in the fungiform and palate papillae, while rarely distributed in the circumvallate and foliate papillae. T1R2: T1R2s are abundantly distributed in the circumvallate and foliate papillae, small in number in the palate, and rare 66

67 in the fungiform papillae. T1R3: T1R3s are ubiquitously distributed in all four types of papillae. Their results suggest that if umami is sensed by hetero-dimers of T1R1 and T1R3, umami should be perceived by mainly by fungiform and palate papillae. Moreover, their results suggest that if sweetness is sensed by hetero-dimers of T1R2 and T1R3, sweetness should be perceived by mainly by circumvallate and foliate papillae, which are localized in posterior portions of the tongue close to the throat. These results suggest the following: (1) if umami molecules bind to the heterodimers of T1R1 and T1R3, umami should be mainly sensed by the fungiform and palate papillae; and (2) if sweetness molecules bind to the hetero-dimers of T1R2 and T1R3, sweetness should be mainly sensed by the circumvallate and foliate papillae, which are localized in the posterior portions of the tongue. However, these results are inconsistent with sensitivity tests on human s tongues investigated by Kieshow, Collings, and others. When we eat ice cream, for example, we lick the ice cream with the tip of the tongue, thus maximizing the sense of sweetness. It means that we know that the tip of our tongues is quite sensitive to sweetness. There are condensed distributions of fungiform papillae on the tip of the tongue. Nevertheless, Nelson et al. (2001) suggested that the heterodimers of T1R2 and T1R3 are only seldomly observed in fungiform papillae. Some more research should be conducted to solve these conflicts. In Conclusion: In this review, we summarized the recent progress on taste research focusing on taste receptors, in particular T1R and T2R families. We described five taste elements, sweetness, bitterness, sourness, saltiness and umami. The receptors of sweetness, bitterness and umami are seven transmembrane proteins coupled with G-proteins, while receptors of sourness and saltiness are ion channels. Sweetness receptors were categorized as T1Rs and bitterness receptors as T2Rs. Heterodimers of T1R1 and T1R3 are responsible for umami, and heterodimers of T1R2 and T1R3 are for sweetness. Their localizations and relation to sensitivities to various taste elements have been also discussed. 67

68 & Figure 1 Bovine taste buds stained with anti-alpha gustducin antibody 68 References Brand, J.G., Cagan, R.H. and Teeter, J.H Receptor events and transduction in taste and olfaction. (Chemical Senses, vol. 1) Marcel Dekker Inc. Chandrashekar, J., Mueller, K.L., Hoon, M.A., Adler, E., Feng, L., Guo, W., Zuker, C.S. and Ryba, N.J T2Rs function as bitter taste receptors. Cell 100(6): Filer, L.J.Jr., Garattini, S., Kare, M.R., Reynolds, W.A. and Wurtman, R.J Glutamic Acid: In: Advances in biochemistry and physiology, Raven Press. Kitagawa, M., Kusakabe, Y., Miura, H., Ninomiya, Y. and Hino, A Molecular genetic identification of a candidate receptor gene for sweet taste. Biochem Biophys Res Commun. 283(1): Kugino, K., Kaneko, M., Akiyoshi, T. and Mizunuma, T Studies on the taste perceptive threshold for 4 basic taste qualities at various sites of lingual surface. Fukuoka Igaku Zasshi 88(10): Li, X., Li, W., Wang, H., Cao, J., Maehashi, K., Huang, L., Bachmanov, A.A., Reed, D.R., Legrand-Defretin, V., Beauchamp, G..K. and Brand, J.G Pseudogenization of a sweet-receptor gene accounts for cats indifference toward sugar. PLoS Genet. 1(1): Margolskee, R.F Molecular mechanisms of bitter and sweet taste transduction. J Biol Chem., 277(1): 1-4. Max, M., Shanker, Y.G., Huang, L., Rong, M., Liu, Z., Campagne, F., Weinstein, H. and Damak, S., Margolskee, R.F Tas1r3, encoding a new candidate taste receptor, is allelic to the sweet responsiveness locus Sac. Nat Genet. 28(1): McLaughlin, S.K., McKinnon, P.J. and Margolskee, R.F Gustducin is a taste-cellspecific G protein closely related to the transducins. Nature 357(6379):