Determination of Antigenic Characteristics and Stabilities of <i>Mycoplasma hyopneumoniae</i>

Weng, Chung-Nan; Lin, Shan‐Yang; Tzan, Y. L.; Lee, C. J.

doi:10.1021/bp00007a012

Cited by 5 publications

(3 citation statements)

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“…The protein level in each sample was determined using a commercial bicinchoninic acid protein assay kit (Pierce, Rockford, IL, USA) with bovine serum protein as a standard according to the manufacture's recommendations. Serum anti-SEP titre was assayed using the ELISA method as Weng et al (1991) described. Briefly, each well of microtitre plates was coated with 100 ll of Mypravac suis lysate solution at a concentration of 1 lg.…”

Section: Plasma Bile and Mucosa Characteristicsmentioning

confidence: 99%

Effects of diets supplemented with organic acids and nucleotides on growth, immune responses and digestive tract development in weaned pigs

Lee

Liu

Chen

et al. 2007

Animal Physiology Nutrition

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Sixty-eight (Experiment 1, 46 days feeding) and sixteen (Experiment 2, 21 days feeding) 21-days-old weaned pigs were allotted to four dietary treatments including control, 0.6% organic acids (OA), 0.1% nucleotides (NA) and 0.6% OA plus 0.1% NA for determining the dietary effects. In Experiment 1, OA enhanced peripheral blood mononuclear cells proliferation on day 28 and 46. The plasma immunoglobulin (Ig) A level was elevated by OA (p < 0.06) and NA (p < 0.07), respectively. In Experiment 2, NA increased plasma IgM level, and had an interactive effect with OA on ileal Peyer's patches and mesenteric lymph node lymphocyte proliferation, bile and plasma IgA levels, and jejunal crypt depth. NA elevated gastric pepsin and jejunal alkaline phosphatase activities, however, decreased ileal aminopeptidase N, sucrase or maltase activity. These results suggest that OA and NA have synergistically enhanced the gut-associated lymphocyte responses and NA modulates the digestive tract development of weaned pigs.

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Section: Plasma Bile and Mucosa Characteristicsmentioning

confidence: 99%

Effects of diets supplemented with organic acids and nucleotides on growth, immune responses and digestive tract development in weaned pigs

Lee

Liu

Chen

et al. 2007

Animal Physiology Nutrition

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“…Indirect ELISA tests were performed as previously described. 25 For coating, MHA was diluted 50-1000-fold in carbonate-bicarbonate solution, pH 9.6. The uncoated surfaces on the plates were sealed by adding 100 pL of 3% (wthol) skim milk (Difco) solution in PBS.…”

Section: Assay Used For Testing Mha Antigenicity (Elisa Test)mentioning

confidence: 99%

Antigenic characteristics and stability of microencapsulated Mycoplasma hyopneumoniae vaccine

Lee

Tzan

1992

Biotech & Bioengineering

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The in vitro stability (temperature, pH, and trypsin) of Mycoplasma hyopneumoniae antigen (MHA) with and without enteric-coated microencapsulation were examined. Microencapsulation of MHA with cellulose acetate phthalate (CAP) is an effective route to produce enteric-coated vaccine microspheres for oral administration. The effect of temperature on the rate of inactivation of MHA was studied by exposing MHA to various temperatures, such as 25, 37, 50 and 60 degrees C. The MHA microspheres were thermally more stable than that of the unencapsulated MHA. The kinetic parameters were observed to follow an Arrhenius-type temperature dependence. The MHA microspheres were also more stable in acidic regions (pH 1.2-4.0) than that of the free one. The enteric-coated MHA microspheres exhibited an excellent enteric function to prevent acidic degradation. A model similar to the well-known Michaelis-Menten equation was formulated to describe the effect of trypsin on the antigenic degradation of MHA. The equilibrium constant K(A) and the maximum reaction velocity V(m) were obtained from experimental data for both free and microencapsulated MHA. Both K(A) and V(m) values of the microencapsulated MHA were smaller than that of the free one, i.e., the resistance to proteolytic enzyme such as trypsin was enhanced by microencapsulation. The storage stability of enteric-coated MHA microspheres has been satisfactorily prolonged that they could preserve more than 90% of original antigenicity after 30 days, and over 80% of antigenicity of MHA was retained in the microspheres for 95 days when it was stored at 4 degrees C.

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“…Enzyme-linked immunosorbent assay (ELISA): Each well of microtiter plates was coated with 100 µl of M. hyopneumoniae lysate solution at a concentration of 1 µg, in a 0.1 M sodium carbonate/bicarbonate buffer (pH 9.6) and stored overnight at 4°C [27]. After suction, 100 µl of PBS containing 0.5% skim milk was used to block the antigen on the plates, which were then incubated at 37°C for 30 min and washed 5 times with PBST (PBS containing 0.05% Tween 20).…”

Section: Preparation Of Microspheresmentioning

confidence: 99%

Protective Effects of Oral Microencapsulated Mycoplasma hyopneumoniae Vaccine Prepared by Co-Spray Drying Method.

et al. 2003

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ABSTRACT. The efficacy of Mycoplasma hyopneumoniae oral vaccine was investigated in microsphere dosage form. A co-spray drying process was used to apply an encapsulating material, Eudragit L30 D-55, to microspheres containing Mycoplasma hyopneumoniae antigens. The microspheres were generally effective (>93%) with protein release at pH 7.4, but almost none were released at pH 1.2, for 3 hr in an in vitro dissolution test. An SPF-swine model was used to evaluate the effectiveness of the microspheres as an oral vaccine, and the related immune responses. The serum's systemic IgG against M. hyopneumoniae was evoked by ELISA analysis, after a 2nd immunization of all pigs. The vaccinated groups' mean lesion score was significantly lower after the Mycoplasma hyopneumoniae challenge than that of the nonvaccinated/challenged groups (P<0.05). This study strongly suggests that the oral microspheres vaccine prepared by a co-spray drying method can provide effective protection against M. hyopneumoniae infection in pigs. KEY WORDS: co-spray drying, microsphere, Mycoplasma hyopneumoniae, oral vaccine.J. Vet. Med. Sci. 65(1): 69-74, 2003 Mycoplasma hyopneumoniae is widely recognized as a potent pathogen for mycoplasmal pneumonia in swine [2], colonizing the respiratory epithelia and compromising integrity by inducing an inflammatory response [20]. The pathogen also causes significant economic losses by reducing body weight and prolonging periods between feeds [22]. Although some drugs and antibiotics are effective in vitro against M. hyopneumoniae, reducing the disease's clinical signs [12], they have not yet been proven effective in eliminating the pathogen in vivo. Vaccination is an essential strategy in controlling mycoplasmal pneumonia. M. hyopneumoniae infects the ciliated epithelial cells of the respiratory tract [6], and a mucosal immune response may therefore be important in the prevention and control of M. hyopneumoniae-induced pneumonia [23]. The concept of a common mucosal immune system has been supported by several oral immunization reports [8,13]. Numerous experimental systems have proven that oral immunization can induce antibody secretion into the mucus on these surfaces [8,16,18], and is almost completely restricted to the secretory form of IgA. Oral immunization causes this IgA antibody to be secreted from the GALT to distant mucosal tissue, including the salivary, mammary, respiratory, intestine and genital mucosal tissue, protecting these areas from pathogen invasion [5,14].Microspheres have recently received widespread interest as vehicles for the controlled release of drug or bioactive agents; resisting degradation in gastric acid, they can release drugs in the intestines [1]. Oral antigen delivery by pHdependent microspheres can improve local and systemic immune responses [4,16], serving as a potent mucosal immunogen. It is essential to develop a new, simple and effective vaccine process. Microsphere oral vaccines have been previously developed in our laboratory via a novel cospray-drying process [16]. C...

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Determination of Antigenic Characteristics and Stabilities of Mycoplasma hyopneumoniae

Cited by 5 publications

References 5 publications

Effects of diets supplemented with organic acids and nucleotides on growth, immune responses and digestive tract development in weaned pigs

Effects of diets supplemented with organic acids and nucleotides on growth, immune responses and digestive tract development in weaned pigs

Antigenic characteristics and stability of microencapsulated Mycoplasma hyopneumoniae vaccine

Protective Effects of Oral Microencapsulated Mycoplasma hyopneumoniae Vaccine Prepared by Co-Spray Drying Method.

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