• Korean Journal of Veterinary Service
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• v.35 no.2
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• pp.133-137
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• 2012

Brucellosis, a bacterial zoonoses caused by the genus Brucella is responsible for abortion and infertility in cow. Brucellosis is causing economic loss in dairy industries and prevalent worldwide including Bangladesh but limited studies are devoted to determine the prevalence and its association with reproductive factors of dairy cows in Bangladesh. Therefore, the present study was conducted to determine the seroprevalence of brucellosis in dairy cattle using screening test Rose Bengal test (RBT) and the positive sera were further confirmed by indirect- ELISA. For this purpose, a total of 400 serum samples from dairy cows with history of abortion and various reproductive disorders were collected from the Kurigram district of Bangladesh for the detection of Brucella antibody. The overall prevalence of brucellosis in dairy cattle was 2.25%. Brucellosis in cases of abortion and repeat breeding was 8.3% and 2.8%, respectively. The results shows higher prevalence of brucellosis in cases of abortion followed by repeat breeding, while there was no seropositive cases from other reproductive disorders. Age-wise sero-prevalence was found 3.0% in 2~3 years age group and 2.0% in 4~8 years age group. The prevalence of brucellosis in indigenous and cross-bred cattle was 3.6% and 1.7%, respectively. All the animals detected positive to brucellosis by RBT were not found to be positive by i-ELISA. However, the RBT might be a suitable screening test for the diagnosis of Brucella infection in field condition in Bangladesh. These data will help to develop effective disease prevention strategies.

• Parasites, Hosts and Diseases
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• v.33 no.4
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• pp.395-398
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• 1995

A 56 year-old Korean housewife/farmerlgoat keeper suffered from right upper quadrant pain and fever with chills. In the abdominal sonogram and computerized tomography, multiple, 2-3 cm, irregular shaped cavities were observed in the right lobe of liver. A liver biopsy revealed extensive central necrosis with Characot-Leyden crystals surrounded by palisading histiocytes, eosinophil-rich inflammatory infiltration. Worm was not observed. However, the serologic test for Fusciola-specific IgG antibody by micro-ELISA was positive. Prior antibody levels did not differ and eosionophilia persisted 6 and 16 months after praziquantel treatment although the cavitaxy lesions in the liver disappeared 6 months after the treatment. Reported herein is a human case of invasive fascioliasis diagnosed clinically by a combination of radiological, histopathological and serological studies.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.4
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• pp.500-506
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• 2006

An experiment was conducted with twenty crossbred male calves (7-15 days old) divided into 4 different experimental groups on the basis of body weights to study the effect of vitamin E supplementation on the serum ${\alpha}$-tocopherol concentration and immune response of the calves. All the calves were fed on milk and calf starter up to 13 weeks and afterwards, they were fed on concentrate mixture and oat hay up to 32 weeks of age. In addition, the calves in groups I, II, III and IV were supplemented with 0, 125, 250 and 500 IU feed grade DL-${\alpha}$-tocopheryl acetate, respectively. Blood samples were collected at 0 day and subsequently at 1, 2, 3, 4, 6 and 8 months of age to monitor the serum ${\alpha}$-tocopherol concentration in crossbred calves. After 24 weeks of experimental feeding, 4 animals from each group were intramuscularly inoculated with single dose (3 ml) of Haemorrhagic septiceaemia (Pasteurella multocida P52 strain) oil adjuvant vaccine. The cumulative group mean serum ${\alpha}$-tocopherol concentration (${\mu}g/100ml$) was 88.12, 210.11, 235.21 and 294.02 in-groups I, II, III and IV, respectively and differed significantly (p<0.001) among the four groups. Lymphocyte stimulation indices (LSI) did not differ among the groups significantly. The pooled mean ELISA antibody titer against Pasteurella multocida (P52 strain) was 788.02, 926.85, 1,214.00 and 1,109.51 for group I, II, III and IV, respectively, which indicated higher antibody titer in groups supplemented with vitamin E as compared to the control group. It may be concluded that vitamin E supplementation increased the ${\alpha}$-tocopherol concentration in serum and dietary supplementation of vitamin E at higher level has a humoral immune enhancing effect against killed bacterial antigen.

• BMB Reports
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• v.30 no.6
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• pp.390-396
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• 1997

A sandwich-type enzyme-linked immunosorbent assay (ELISA) for the quantification of human apolipoprotein A-I (apoA-I) was developed using monoclonal antibodies. For this assay, we used three monoclonal antibodies to trap and detect apo A-I. HDAI16 and HDA15 monoclonal antibodies were used for trapping apoA-I and HDAI8 monoclonal antibody was for detecting apoA-I. These three monoclonal antibodies were produced by immunizing mice with high density lipoprotein (HDL) isolated from human plasma. By immunoblot analysis, these three monoclonal antibodies were specific to apoA-I and showed no cross-reactivities with other plasma proteins. The results of competition assays for epitope cross-reactivity test also verified that these monoclonal antibodies identified separate and distinct epitopes on HDL and apoA-I. Affinity constants of monoclonal antibodies were measured by ELISA. Their association constants ranged from $10^7$ to $10^8$ $M^{-1}$. For this assay, pure apoA-I was isolated by affinity chromatography using monoclonal antibodies. In this sandwich assay, the amount of HRP-labeled HDAI8 bound to apoA-I trapped by HDAI16 and HDAI5 was proportional to apoA-I concentration in the range of 0 to 500ng/ml. ApoA-I concentration in plasma was calculated from the linear regression equation of standard curve. The precision and reliability of the assays are reflected in the low intra-and interassay coefficients of variation that averaged 3.25% and 4.30%, respectively. This assay is sensitive, simple, reproducible, convenient in incubation interval, and does not use radioisotope: thus it can be widely applied in clinical laboratories.

• Clinical and Experimental Pediatrics
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• v.54 no.4
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• pp.163-168
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• 2011

Purpose: The purpose of this study was to evaluate the immune response to serotype 19A in children aged 12-23 months after immunization of the 19F containing 7-valent pneumococcal conjugate vaccine (PCV7). Methods: Blood samples from a total of 45 subjects (age 12-23 months) were included in the study. Subjects were categorized according to immunization status into three groups as follows: 18 subjects with 3 primary doses and 1 booster dose of PCV7 (booster group), 21 subjects with 3 primary doses before 12 months of age (primary group), and 6 subjects with no vaccination history of PCV7 (control group). An ELISA and opsonophagocytic killing assay (OPKA) was done to evaluate the immune responses against serotypes 19F and 19A. Results: According to the ELISA, all subjects had antibody titers ${\geq}0.35{\mu}g/mL$ for serotypes 19F and 19A in the booster and primary group and 83.0% and 66.7% in the control group, respectively. According to the OPKA, subjects with opsonic activity (${\geq}20$) against serotypes 19F and 19A were 100% and 61.1% of the subjects in the booster group and 66.7% and 19.0% in the primary group, respectively. No subjects in the control group had opsonic antibodies against both serotypes. Conclusion: In conclusion, in children 12-23 months age who were previously vaccinated with PCV7, a cross-reactive immune response is elicited against serotype 19A after a primary series of 3 doses in a small proportion of subjects, and this response is amplified after booster vaccination.

• Korean Journal of Veterinary Research
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• v.43 no.4
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• pp.677-681
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• 2003

Currently, both the indirect fluorescent antibody test (IFAT) and enzyme linked immunosorbent assay (ELISA) have been used to detect Neospora caninum antibodies. Several factors such as the buffers, the conjugate, the pattern of fluorescence, and the cross reactivity with other apicornplexan protozoan, may result in poorly correlated data. The present study was undertaken to develop and evaluate the Neospora agglutination test (NAT) for the detection and quantification of IgG antibodies to N. caninum from various animal species. Compared to the ELISA method, the NAT with a cutoff value of 1:512 gave a high index of coincidence (kappa=0.807) and no cross reactivity to Toxoplasma gondii antiserum. Hence, this NAT method, which did not require a species-specific secondary antibody and expensive tools, would be easily available for the detection of antibodies to N. caninllm of various animal species.

• Journal of Periodontal and Implant Science
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• v.39 no.sup2
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• pp.231-237
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• 2009

Purpose: Cytolethal distending toxin (CDT) is a family of heat-labile cytotoxins produced by several gram-negative mucosa-associated pathogens, including Aggregatibacter actinomycetemcomitans. CDT is well known to be capable of inducing growth arrest, morphological alterations, and eventually death in various cells. CDT belongs to a tripartite $AB_2$ toxin (CdtB: the enzymatic A subunit; CdtA and CdtC: the heterodimeric B subunit). Previous studies proposed that CdtA and CdtC together bind to a cell surface receptor and glycolipids act as a receptor for A. actinomycetemcomitans CDT (AaCDT). In this study, recombinant CdtA and CdtC proteins of AaCDT were co-expressed in a bacterial expression system and tested for their affinity for $GM_1$ ganglioside. Methods: The genes for CdtA and CdtC from A. actinomycetemcomitans Y4 were utilized to construct the expression vectors, pRSET-cdtA and pET28a-cdtC. Both CdtA and CdtC proteins were expressed in Escherichia coli BL21(DE3) and then purified using hexahistidine (His6) tag. The identity of purified protein was confirmed by anti-His6 antibody and monoclonal anti-CdtA antibody. Furthermore, the affinity of recombinant protein to $GM_1$ ganglioside was checked through ELISA. Results: Recombinant CdtA and CdtC proteins were expressed as soluble proteins and reacted to anti-His6 and monoclonal anti-CdtA antibodies. ELISA revealed that purified soluble CdtA-CdtC protein bound to $GM_1$ ganglioside, while CdtA alone did not. Conclusions: Co-expression of CdtA and CdtC proteins enhanced the solubility of the proteins in E. coli, leading to convenient preparation of active CdtA-CdtC, a critical material for the study of AaCDT pathogenesis.

• The Journal of Korean Society of Virology
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• v.26 no.1
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• pp.59-65
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• 1996

For easy and rapid screening of hemorrhagic fever with renal syndrome (HFRS) without any laboratory equipment, dot blot enzyme immunoassay was developed and tried to detect anti-hantavirus antibodies. The nucleocapsid protein of Hantaan virus was isolated by affinity chromatography and used for making the dot strip. 28 of 29 Hantaan virus infected sera showed positive signals and 21 of 22 HFRS negative sera showed no positive signals. Anti-Seoul virus monoclonal antibody also exibited positive signal but the intensity of colorization was approximately 5 fold less than that of anti-Hantaan monoclonal antibody. The sensitivity of dot blot assay was equal or superior to indirect immunofluorescent assay (IFA) or ELISA test. Overall, the screening results with dot blot assay showed 92.2 % of concordance with IFA or ELISA test. This results suggests that dot blot assay could be applied a tool for easy and rapid screening of HFRS.

• Korean Journal of Veterinary Service
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• v.30 no.1
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• pp.51-66
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• 2007

The purpose of this study was to evaluate the effect of a subsequent infection of porcine reproductive and respiratory syndrome(PRRS) virus to pigs with A pleuropneumonia. Twenty three 7-week-old commercial pigs were infected intratracheally with PRRS virus and/or A pleuropneumoniae serotype 5. Serum antibody titers were examined by an enzyme-linked immunosorbent assay(ELISA) and proportion of porcine leukocyte subpopulations in peripheral blood was examined by flow cytometry. In this experiment, antibodies against PRRS virus and A pleuropneumoniae were detected at 2 weeks and 1 week postinfection and the number of antibody positive pigs were gradually increased. And in proportion to leukocyte subpopulations in peripheral blood of pigs infected with A pleuropneumoniae compared with pigs administrated with saline, the proportion of PoCD4 and N cells were increased(P<0.1). Furthermore, in proportion to leukocyte subpopulations in peripheral blood of pigs infected with PRRS virus followed by A pleuropneumoniae compared with pigs administrated with saline, the proportion of MHC class II, PoCD4 and B cells were significantly increased(P<0.1). The results indicated that dual infection with PRRS virus and A pleuropneumoniae induced the stronger immune responses associated with macrophages and Th cells in pigs than single infection with PRRS virus or A pleuropneumoniae.

• Biomedical Science Letters
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• v.8 no.4
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• pp.235-244
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• 2002

This study was carried out to determine the blood and milk progesterone by enzyme-linked immunosorbent assay (ELISA), and milk urea nitrogen (MUN) in cows. MUN and protein concentration were determined using automated infared procedures. The optimum conditions of ELISA system was investigated including the first and second antibody titres, bound percent, and enzyme conjugate and also the factors on MUN and protein concentration by sampling procedures and addition of preservatives. Progesterone antibodies did not react to pregnenlone, testosterone, estrone, estradiol-l7$\beta$, aldosterone, cortisol, corticosterone and 11$\alpha$-dehydroxycortisone (DOC), but reacted with only progesterone. The intra and inter-assay coefficient of variation 4.5%, 6.1~9.4% when used of bovine serum. The morning, MUN concentration (17.6$\pm$2.8 mg/100 ml) in the 13 herds was similar to that of evening MUN concentration of the lactating cows from the same herd. A significant relationship between morning and evening milk samples of upper parameters was found r=0.93. Difference in MUN concentration with sampling procedures and using of preservatives were investigated.