ABSTRACT
Lactobacillus spp. inhibit the growth of Campylobacter spp. in vitro. However, in chicken crops, in which Lactobacillus spp. predominate, such inhibition of Campylobacter has not been confirmed. In our previous study, feeding paddy rice to broiler chicks increased the residence time of the food, which might enhance the bactericidal activity of the crop. Here, the bactericidal activity against the remaining Campylobacter spp. in broiler crops was evaluated. A suspension prepared by mixing Campylobacter jejuni and titanium dioxide (TiO2) was inoculated into the pharynx of 26-day-old broiler chicks fed a paddy rice-based diet. The crop contents were sampled at 20-min intervals. The TiO2 residual ratio in the crop gradually decreased with time after inoculation, with 57% of the inoculated TiO2 remaining in the crop 60 min after inoculation. The survival fraction of C. jejuni in the crops was 11% at 40 min, only 1% at 60 min, and was undetectable at 80 min. Most of the inoculated C. jejuni died in the crop before entering the next segment. These data indicated that bacterial death occurred between 30 min and 40 min after inoculation. The average survival time of C. jejuni in the crop was calculated to be 37.1 min. Thus, C. jejuni remaining in a chicken crop for more than 40 min died.
ABSTRACT
DNA immunization has been used to study vaccination methods and for production of specific antibodies. The present study aimed to apply DNA immunization to prepare specific IgYs, which react against rabies virus N protein (RV-N) and can be used to research and diagnose rabies virus. The DNA sequence of RV-N was ligated into a pcDNA 3.1 plasmid for constructing pcDNA-N. Eight hens were divided into four groups. Group 1 comprised the control group (non-immunized). In Groups 2, 3, and 4, hens were injected intramuscularly with pcDNA-N (400 µg/hen). Eight injections were administered every other week. From the 4th week, an adjuvant was injected in addition to pcDNA-N. Freund's complete adjuvant (FCA) and λ-carrageenan were administered to Groups 3 and 4, respectively. Eggs were collected daily, and the specific antibody activities of egg yolks were measured by ELISA. IgYs were purified from pooled egg yolks at 16-19 weeks post-administration in each group. The detection sensitivities of the RV-N were compared using purified IgY as the primary antibody for ELISA, dot blotting, and western blotting. Egg yolks from one of the two hens in Group 2 (pcDNA-N alone) and all hens in Groups 3 (pcDNA-N + FCA) and 4 (pcDNA-N + λCarra) had increased ELISA values. The combined use of λ-carrageen in DNA immunization resulted in an adjuvant effect comparable to that of FCA. Each purified specific IgY detected RV-N in the ELISA, western blotting, and dot blotting; however, the detection sensitivity differed. Higher detection sensitivity of the +λCarra IgY was observed by ELISA, whereas there was higher detection sensitivity of +FCA IgY in western blotting and dot blotting. In summary, anti-rabies virus N protein IgY was prepared through DNA immunization of hens using FCA or λ-carrageenan as adjuvants and can be used as a primary antibody to detect rabies viruses.
ABSTRACT
Objectives: Atopic dermatitis (AD) is one of the most common skin disorders in infants and children and is often aggravated by increased Staphylococcus aureus (S. aureus) colonization. An inhibitory effect of a specific egg yolk antibody (IgY) on S. aureus growth was demonstrated in this study. Furthermore, the effects of water- or oil-based adjuvants on the preparation of anti-S. aureus IgY and hen immunization were compared. Methods: Hens were immunized intramuscularly with formalin-killed S. aureus mixed with either a water-soluble polysaccharide λ-carrageenan, oil-based Freund's complete adjuvant (FCA), or Freund's incomplete adjuvant (FIA). Anti-S. aureus IgYs (FIA-IgY, FCA/FIA-IgY, and λCarra-IgY) were purified from the egg yolk of immunized hen eggs, and the activity of the IgY against S. aureus antigen was measured by ELISA. The proportion of each IgY that was absorbed by S. aureus was also determined. Then, the effect of purified anti-S. aureus IgY on S. aureus growth inhibition was investigated in vitro. Results: The yolk of eggs and purified FIA-IgY from the FIA group showed the highest antibody activity, followed by FCA/FIA-IgY and λCarra-IgY. The proportion of each IgY that was absorbed by S. aureus antigen was as follows: FIA-IgY (18.1%), FCA/FIA-IgY (12.9%), and λCarra-IgY (7.0%). Only FIA-IgY significantly inhibited S. aureus growth in liquid medium. Conclusion: A specific IgY that was produced using the FIA adjutant inhibited S. aureus growth. Although water-soluble λ-carrageenan showed an adjuvant effect on anti-S. aureus IgY induction in egg yolk, but did not inhibit S. aureus growth. The use of the oil adjuvant FIA was necessary in the preparation of anti-S. aureus IgY as a treatment for AD symptoms.
ABSTRACT
We previously reported that a diet containing 65% paddy rice suppressed the colonization of Campylobacter jejuni in the cecum of broiler chicks, suggesting that this type of diet has positive effects on upper gastrointestinal tract function in broilers. To reveal the possible mechanisms involved in this antibacterial effect of the whole-grain paddy rice diet, we performed experiments comparing the digesta passage rate in the crop and gizzard, the development of gizzard, and the pH distribution in the gizzard between groups of chicks fed two different diets, such as ground corn and whole-grain paddy rice. During these experiments, we made the following observations: the chicks in the group fed the whole-grain paddy rice diet had more developed gizzards and significantly larger crop content than the chicks in the group fed the ground corn diet. The chicks fed the whole-grain paddy rice diet retained the digesta in the crop for much longer and had less variation in the pH values in the gizzard than those fed ground corn. On the basis of these observations, we concluded that the hardness of the rice hull in whole-grain paddy rice leads to a larger amount and longer retention of content in the crop, as well as the uniformity of the internal pH of the gizzard, by promoting gizzard activity. We speculate that the hardness of the rice hulls promoted the grinding activity of the gizzard, resulting in the long retention time of the digesta in the crop and uniformity of the internal pH of the gizzard, which may sterilize or suppress Campylobacter growth in the gastrointestinal tract of broiler chicks.
