Passive protective effect of egg yolk antibodies

against some animal diseases

I. INTRODUCTION
Livestock agriculture still plays an important role in the economy. Therefore, diseases of
animals are always a concern for farmers, they may be empty-handed due to their
animals’ illness... That worry has subsided since the birth of antibiotics. Antibiotic use
significantly improves the animal’s performance and their health status. However, the
use and misuse of in-feed antibiotics have led to problems of drug residues in animal
products and increased bacterial resistance. As a result, many countries, including
Vietnam, have been enacting a ban on antibiotics in food. Therefore, alternatives to
antibiotics are urgently needed.

Recently, chicken egg yolk immunoglobulin, referred to as immunoglobulin Y (IgY) has

attracted considerable attention as a means to prevent and control the disease. Egg yolk
provides a more hygienic, cost-efficient, convenient, and rich source of antibodies
compared with the traditional method of obtaining antibodies from mammalian serum.
Specific antibodies were made by specific antigenes (Vibrio parahaemolyticus, Vibrio
harveyi, Vibrio anguillarum) were injected into the hen’s breast. After that, it will switch to egg
yolk and then produce IgY. In contrast to antibiotics, the use of IgY is environmentally-
friendly and elicits no undesirable side effects, disease resistance, or toxic residues
(Coleman, 1999).

To clarify the therapeutic effects on animals of IgY, in this study, we analyzed two
subjects: pigs with diarrhea caused by E. coli bacteria and shrimp with white spot
syndrome due to white spot syndrome virus (WSSV).

II. MATERIAL AND METHODS

2.1. Research subjects

2.1.1. Diarrhea in pigs caused by E. coli

Twenty 21-day-old healthy Cotswold piglets were randomly divided into two groups, i.e.,
the control and the treatment, of 10 piglets each. All piglets were challenged twice each
 12 -1
with 5 ml of E. coli K88 + MB at a dose of 10 CFU ml per piglet. The suspension or
E. coli K88+ MB was administered at 0 and 5 h of the experiment. After the challenge,
the piglets in the treatment group were treated with 0.5 g of egg-yolk antibodies at a titer
of 140000 (100000-168000) three times a day at times -1, 4, and 9 h each for two
consecutive days after the first E. coli challenge, whereas the control received placebo
treatment. The clinical response of each piglet was monitored throughout the experiment
in terms of occurrence of diarrhea, fecal consistency score, weight loss, and mortality.
(Marquardt et al. 1999)
2.1.2. White spot syndrome virus (WSSV) in shrimp

Healthy Metapenaeus Ensis shrimp (n=15, approximately 10g body weight) were
collected from the market and had continued to be grown under appropriate
conditions.

The dilution of WSSV used in the preliminary challenge test was 100x dilution of crude
WSSV stock.

In the assay, 250 mg/ml IgY powder was incubated with WSSV for 1 h at room
temperature before injection. Then the mixture of antibody and WSSV was
intramuscularly injected into shrimp in the fourth abdominal segment. Preimmune IgY
was used as the control for immune IgY. At the same time, a positive control (WSSV
only) and negative control (0.01 M PBS only, pH 7.4) group were injected. The shrimp
were checked for survival rates twice a day. (Lu et al. 2008)

III. RESULTS AND DISCUSSION

3.1. Diarrhea in pigs caused by bacteria E. coli

Table 1. Clinical response of 21-day-old pigs after challenge with ETEC K88+ MB and
treatment with egg-yolk antibodies.

*Note:

a: The object had been specifically presented in section 2.1.

b: FC score is the mean fecal consistency score: 0, normal; 1, soft feces; 2, mild diarrhea;
3, severe diarrhea. Pigs with a fecal score of ≤ 1 were considered not to have diarrhea.

(From (Marquardt et al. 1999)).

There is a big difference between the two subjects. The control piglets (treated with IgY
from unvaccinated hens) showed increasing and increasingly severe diarrhea.

After 24 hours, up to 30% began to have soft stools, and after 48 hours up to 80% of pigs
had mild diarrhea. From 48h-72h up to 30% of pigs died of diarrhea. Moreover, after 72
hours, up to 57% of pigs had severe diarrhea. Pigs' weight was reduced by 36.2g due to
diarrhea.

In contrast, pigs given IgY from vaccinated chickens had very positive results. However,
after 24 hours, up to 50% of pigs showed signs of soft stools, near mild diarrhea.
Nevertheless, on the second day, the pigs quickly recovered and on the third day, the pigs
remained healthy. Not only did no pig die from diarrhea, but even this group of pigs also
had a slight weight gain of 90.6g.

It can be seen that IgY has a good effect in treating diarrhea in piglets. However, the
number of pigs in this study was not large, so it would be expected to repeat this study
in a greater number of animals.

3.2. White spot syndrome virus (WSSV) in shrimp

 Fig.1. Effect of IgY on the survival rate of shrimp challenged with white
spot syndrome virus.

(From (Lu et al. 2008))

The graph shows that the test subjects tend to reduce survival rates over time.

Shrimp without white spot syndrome (PBS) had the highest survival rate, after 10
days still survived up to 80%.

For shrimp group with white spot syndrome (WSSV):

+ The shrimp group tested with inactivated virus vaccine - IgY (inactivated) had
the highest survival rate, after 10 days still up to 71% of alive shrimp

+ After that, the shrimp group was tested with DNA vaccine, but the survival rate had
dropped sharply to just over 30%.

+ For the remaining 2 groups that were not vaccinated and preimmune IgY, there was no
The 10-day survival rate, which suggested preimmune IgY did not neutralize the WSSV.
Thus, IgY antibodies were effective in treating white spot disease in shrimp. However,
traditional inactivated vaccines still had an advantage in producing antibodies over
DNA vaccines

IV. CONCLUSION

This research had shown two of the numerous animal diseases that have been
improved by IgY. In addition, it is no denying that IgY is also effective in treating
many other diseases, not only in animals but also in humans. This is indeed a
technology that deserves to be exploited and developed.

Furthermore, IgY has many advantages such as humanity, easy collection and high
productivity, and cost-saving. We do not need to sacrifice the human body, no need to or
even kill the animals to produce antibodies. More importantly, IgY is non-toxic, non-
allergic, no serological cross-reactivity.

However, IgY also has some limitations because of its susceptibility to proteolysis. IgY
is fairly resistant to digestion by intestinal proteases (Hatta et al. 1993). It was found that
the activity of IgY was decreased at pH 3.5 and completely lost at pH 3.0 (Shimizu et al.,
1988). IgY has a serum half-life of 1.85 days in newborn pigs. This is shorter than the
reported serum half-life of 12 to 14 days for homologous IgG (colostral antibodies) (Xu
et al. 2011)

It is hoped that we will find new solutions to make IgY technology even more complete.
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