蛔蟲및 鉤蟲의 感染이 宿主의 血淸蛋白分劃像에 미치는 影響에 關한 實驗的硏究

Abstract

[한글]

[영문]

Since the introduction of electrophoretic technique by Tiselius in 1937, Kunkel and Tiselius (1951) proposed the modified technique, paper electrophoretic method. This simple technique has been used in the study of helminthic infestation ever since (Stauber, 1954).

Mori (1955), Baba et al. (1956), Hara(1956), Ishizaki et al. (1956, 1959) and Ashida(1959) reported serum protein changes in hookworm disease but there are few studies in relation to ascaris infestation (Dalin, 1962).

Kang (1959) reported increased susceptibility to Clonorchis sinensis and damage of liver in mice fed nonprotein diet only and Yanagihori (1956) stated an inhibition of ascaris infestation in the mice fed with high protein diet.

Venkatachalam and Patwordham (1953) noticed the decrease of fecal nitrogen after the elimination of ascaris. The present study was designed to study serum protein changes in animals infested with Ascaris lumbricoides, Toxocara canis, and Ancylostoma caninum

(1) in abnormal hosts, during somatic migration of larvae

(2) in normal hosts

(3) in protein fed animals.

And also this study includes the fecal nitrogen excretion in normal and abnormal hosts with or without parasitic infestation.

Materials and Methods

Parasites: Ascaris lumbricoides, Toxocara cains, and Ancytostoma caninum were used as experimental parasites. Eggs of A. lumbricoides and T. canis were taken from the uterus of female adult worms and were incubated at 26-28℃ for 4 to 5 weeks, by Tile culture method. Larvae of A. caninum were obtained from a dog's stool and were cultured for 7 days at 25 to 30℃ (Lee and soh's method, 1964) Eggs and larvae were given to animals by mouth.

Animals: Rabbits, dogs and rats born naturally grown in isolation and proved to be parasites free in their digestive system by Formalin-ether centrifugal method(M.G.L.), were used as experimental animals.

Experimental animal groups were as follows:

1. Abnormal host (Rabbits) groups

Group No. of Rabbits Parasites

1 5 Ascaris lumbricoides 10,000 eggs

2 5 Toxocara canis 10,000 eggs

3 5 Ancylostoma caninum 10,000 larvae

4 5 Normal control

2. Normal host(Dogs) groups

Group No. of Dogs Parasites

1 3 Toxocara canis 100 eggs

2 3 Ancylostoma caninum 100 larvae

3 3 Normal control

3. Protein diet Rat groups

Group No. of Rats Protein content Parasites

1 5 high protein diet(Casein 25%, Rice 65% A. lumbricoides,

10,000 eggs

2 5 low protein diet(casein 15%, Rice 75%) A. lumbricoides,

10,000 eggs

3 5 infectious control(casein 0%, Rice 90%) lumbricoides, 10,000

eggs

Table of protein diet of infectious controls.

Casein 0%

Rice 90%

Olive oi1 4%

Salt mixture 4%

Vitamin mixture, Cod liver oil and Yeast 2%

Serum collection method: From rabbits and dogs, the serum was collected after 15 minutes centrifugation at 3,000 r.p.m. with 2 to 3 ml. of blood which was placed at room temperature for 20 minutes. From rats the blood was taken from the cut tail with capillary tube and the serum was obtained after 5 minutes centrifugation at 11,000 r.p.m. by Adams autocrit.

Total serum protein was measured by the Micro-Kjeldahl method and the serum protein fraction was measured using paper electrophoresis in which serum was run on the Whatman No.1 filter pater for 16 hours at 70 volt and the reading done by Junior spectrophotometer at 520 mμ.

Total fecal nitrogen was measured by Folin-Farmor Micro-Kjoldahl method.

Results

Serum protein changes in abnormal hosts infested with Ascaris lumbricoides, Toxocara canis, and Ancylostoma caninum. In the group infected with Ascaris lumbricoides there was gradual increase of total protein and at the end of 3 weeks the protein level was higher by 1.0 gm/dl than in the control group. Whereas the albumin level was decreasing, and at the end of 7 weeks there was about 20% decrease, there were little differences in α^^1-and, α^^2-globulin in comparison to normal controls. There was 4 to 10% increase of β-globulin in 5 to 7 weeks and 10% increase of γ-globulinin 3 to 5 weeks in experimental group. In the groups infested with toxocara and ancylostoma the results were similar to the ascaris group in that in all infested groups there were serum protein changes during the somatic migration of larvae and at the end of 5 weeks the changes showed recovery toward normal.

Serum protein charges of normal hosts infested with Toxocara canis and Ancylostoma caninum. In the group infested with toxocara, the total protein level was similar to that of control group. However there was a gradual decrease of serum albumin and at the end of 7 weeks there was a 17% decrease in the albumin level. α^^l-, α^^2- and β-globulin levels were similar to those of normal controls but at the end of 3 weeks there was a marked increase of γ-globulin (30%), The A/G ratio, therefore, had changed from 0.43 to 0.33.

In the group infested with ancylostoma the results ware similar to those of the toxocara infested group. However, the total protein level showed some increase at the end of one week and then a decrease of 0.7 gm/dl in comparison to normal controls.

Therefore in the abnormal host there was a trend to recovery toward normal at the end of 5 weeks, whereas in normal hosts there was a continuation of changes in serum protein level.

Serum protein changes in protein fed abnormal host infested with Ascaris lumbricoides. The body weight in experimental groups was 197gm in low protein diet fed group and 216 gm in high protein diet fed group in comparison to normal control of 146 gm, after 50 days feeding, prior to the infestation with Ascaris

lumbricoides. Therefore the increase ratio of body weight were 2.02 gm/day in control, 2.95 gm/day in low protein fed group and 3.39 gm/day in high protein fed group. Serum albumin was decreasing at the end of 5 weeks(45.1±2.06%) but α^^1-,

α^^2- and β-globulin were little different in comparison with preinfestation level. r-globulin was higher by 6.4% in comparison to preinfestation level of 11.6±1.92%. At the end of 5 weeks the A/G ratio dropped to 0.83 from 1.18 and was similar to that of the control group.

Fecal nitrogen excretion in hosts infected with parasites. In the dog, the normal host, the excretion was 7.0 gm/dl in control group. However, after 5 to 7 weeks of infestation the total amount of nitrogen excretion ranged to 7.7-8.0 gm/dl. In the rat, the abnormal host, the nitrogen was increased at the earlier stage but after 5 weeks there was a trend of recovery toward normal. In the group fed with protein diet the nitrogen ecretion was increased in comparison with normal controls.

Conclusion

The influence of the infestation of Ascaris lumbricoides, Toxocara canis, and Ancylostoma caninum in normal, abnormal, and protoin fed host upon serum electrophoretic changes and fecal nitrogen excretion.

1. During the somatic migration of larvae of Ascaris lumbricoides, Toxocara canis, and Ancylostoma caninum in the abnormal host there was little changes in α^^1-and α^^2-globulin, but total serum protein, pβ-and γ-globulin showed an

increase and albumin and A/G ratio showed a decrease. After the somatic migration the earlier changes of protein electrophoresis showed a trend of recovery toward normal.

2. In the normal hosts infested with Ascaris lumbricoides and Ancytostoma caninum the serum electrophoretic changes showed a marked decrease of albumin and A/G ratio in comparison to normal controls. However, the total serum protein and γ-globulin

level were increased after infestation with ascaris and hookworm.

3. In abnormal hosts fed with protein diet total serum protein, fecal nitrogen content, and γ-globulin level were increased at the earlier phase of ascaris infestation, whereas albumin and A/G ratio were decreased. Howeever, there were no remarkable differences in comparison to infectious controls fed with non-protein diet.

4. The total fecal nitrogen content in normal hosts infested with Ascaris lumbicoides and Ancylostoma raninum showed a marked increase in comparison to normal controls after 3 weeks. In abnormal hosts infested with Ascaris lumbricoides there was an increase of fecal nitrogen content up to 3 weeks but was a decrease after 5 weeks.

From above results we conclude that in ascaris and ancylostoma infestation, during the period of somatic migration of larvae and adult atage form, the total serum protein and γ-globulin level increase, albumin and A/G ratio decrease, and excretion of fecal nitrogen increases.