ABSTRACT
Traditionally, acupuncture (AP) points used to treat respiratory disease include SanTai, FeiYu, An-fu and SuQi. Clinical, bacteriological, virological and immunological effects of AP were studied in calves with respiratory disease, which were allocated to one of 5 groups for treatment by: 1=Medicine alone; 2=Medicine + AP at SuQi; 3=Medicine + AP at (SanTai + FeiYu + AnFu); 4=AP at SuQi alone; 5=AP at (SanTai + FeiYu + AnFu) alone. In the AP groups, needles were inserted into the points to a depth of 3 cm for 20 minutes daily for three days.
Clinical recovery rates (Excellent to Good) were respectively: 1=59%; 2=83%; 3=73%; 4=83% and 5=63%. Virus isolation rate was significantly decreased in calves with viral respiratory diseases after AP. Otherwise, antibody response of calves given AP, especially at SuQi, was increased significantly. The percentages of leucocyte subpopulation expressing MHC class II antigen, CD2, CD4, sIgM and N12 antigens were significantly higher in the calves with respiratory disease treated with AP.
AP therapy reduced the clinical signs in calves with respiratory disease and promoted their immune responses. Therapy which combines medicine with AP could be put to practical use in field outbreaks of respiratory disease in calves.
KEYWORDS
Acupuncture, Bronchopneumonia, Immune response, Leucocyte subpopulation, Respiratory disease, Virus isolation rate.
INTRODUCTION
Infectious respiratory disease is a serious threat to the profitability of animal husbandry, especially in intensive units. The main infectious agents involved are viral (especially IBR, RSV, PI3, BVD) and bacterial (especially Pasteurella). Calf pneumonia results in great economic loss. The prevalence of respiratory disease in calves increases as herd size increases. Therefore, respiratory diseases are attributable to multiple factors, which include management and environment, etc. The major clinical signs of bovine influenza include fever, cough and nasal discharge. Fever, cough and dyspnea are common in bronchitis. When bronchitis develops to pneumonia, lung rales (rassel-sounds) are easily found and the condition is more serious1.
In Western countries, bovine respiratory disease often arises in spite of the use of currently available vaccines. Therapy of clinical cases often uses medication with antibiotic- and/or antiinflammatory- drugs (such as corticosteroids), bronchodilators and antitussives. Except to control secondary bacterial infection, antibiotic therapy seldom helps in viral diseases. Steroid therapy is problematical; it can aggravate the clinical signs and increase the mortality rate. To avoid the use of steroids, non-steroidal antiinflammatory drugs (NSAIDs)5 and antiprostaglandins have been recommended6. Antiprostaglandins effectively decreased respiratory rates, lowered body temperature and stopped cough in calf pneumonia6.
The prevalence of calf respiratory disease, especially pneumonia, has been investigated recently in Korea. The prevalence rate ranged from 13-64% and many calves died. Respiratory disease was more prevalent during the suckling and growing period, especially in calves less than one month old. Mortality rate was highest at that age. Although various approaches have been tried to reduce the occurrence of bovine respiratory disease, the disease is still troublesome.
In Eastern countries, herbal medicine (especially Traditional Chinese Medicine (TCM)) and AP have been used for centuries to treat respiratory disease7,15,16. For example, AP is used to reduce fever, to increase phagocytosis and local immunity in the lung, bronchi and trachea, to dilate the bronchi, to enhance mucolysis and expectoration in cough, to enhance circulation and to increase the supply of vital nutrients7,16. As it can support the internal organs and increase their defensive reactions, AP therapy can reduce the dose and the frequency of medication7. AP combined with western therapy was effective in treating respiratory diseases7. In 21 calves with bronchitis, TCM gave excellent and good results in 62% and 33%; 5% were not cured3. AP significantly decreased tracheal resistance of 9/12 patients with bronchial asthma at 10 minutes, 1 hour and 2 hours after AP treatment2. Over 70% of bronchial asthma patients were significantly improved at 10 weeks after AP treatment8. AP in rats had some preventive effects on damage to tracheal epithelial cells and enhanced mucosal secretion12. In mice, AP stimulation (especially, by filiform needle (hao chen) and by hot needling) had positive effects on the immunosuppressive response caused by cold stimulation. The most effective point to improve the immunosuppressive response caused by cold stimulation in mice was Hou-san-li (ST36)13,14.
As antimicrobial agents are not very effective (especially in viral diseases) and consumers demand food free of chemical residues, we decided to study the efficacy of AP to treat calves with clinical respiratory disease. Our aim was to develop the most effective therapy for respiratory disease in calves and ultimately to improve their productivity. Different authors used different AP points to treat respiratory disease in calves: TaiMai and AnFu (Klide and Kung4); FeiYu and BiYu (Kurosawa & Sakai11); ShanKen, BiYu, TaiMai, FeiNei, SanTai, SuQi and FeiYu (Yang and Lee9); SuQi, AnFu and FeiYu (Woo10).
Using 89 calves with clinical respiratory disease, we compared the cure rate of medicine alone, medicine combined with AP (at a point complex (SuQi), or at a combination of three points (SanTai + AnFu + FeiYu)) and AP alone (at the same points). We also examined some aspects of the immune responses of treated calves.
MATERIALS AND METHODS
Patients and treatments: Calves with respiratory disease (influenza, bronchitis, pneumonia and bronchopneumonia) were submitted to the trial from local rearing farms. The respiratory diseases were not differentiated. Clinical cases (n=89) were selected and were allocated to one of five treatment groups, as follows:
| Number of calves treated | ||||
| Group | Treatment | Spring | Summer | Overall |
| 1 | Medicine only | 5 | 12 | 17 |
| 2 | Medicine + AP at SuQi | 10 | 8 | 18 |
| 3 | Medicine + AP at (SanTai + FeiYu + AnFu) | 5 | 10 | 15 |
| 4 | AP at SuQi only | 15 | 8 | 23 |
| 5 | AP at (SanTai + FeiYu + AnFu) | 7 | 9 | 16 |
| TOTAL | 42 | 47 | 89 | |
The medicine used was mainly injection of antibiotic (enrofloxacin), analgesic-antipyretics and antitussives, as required.
In groups 2 to 5, round sharp AP needles were inserted into the selected points to a depth of 3 cm, and remained in situ for 20 minutes. The angular direction of needling was forwards and downwards between the vertebral spines of the three midline points (AnFu, SanTai and main SuQi point) and inwards and downwards into the muscles at the other points (FeiYu and secondary SuQi points).
Sampling: Blood and sputum were obtained aseptically from the patient on the day before treatment, and on day 1, 3 and 10 posttreatment. The samples were transferred to National Veterinary Research Institute (Anyang, Korea) in order to investigate the infectious agents and to pursue the changes of host cellular immune response after AP application.
Evaluation of therapeutic effects: Clinical recovery was evaluated by four different grades as follows:
Excellent: positive effects +++
Good : positive effects ++
Fair : positive effects +
Poor : no, or negative, effects
Virus isolation and viral seroconversion rate: Cells were propagated in a -Minimum Essential Medium (a -MEM, GIBCO) supplemented with 7% fetal calf serum (FCS). Buffy coat from the blood and nasal secretion were cultured with a monolayer of Mardin Darby bovine kidney (RK-13) at 37oC and the cytopathic effects were observed. Bovine viral diarrhoea virus (BVDV) showed cytopathic effect in only RK-13 cells but Infectious bovine rhinotracheitis virus (IBRD) and Parainfluenza virus-3 (PI3) showed cytopathic effect in both MDBK and RK-13 cells. The infectious cells were fixed with cold acetone after washing with phosphate-buffered saline and identified by the direct fluorescent antibody technique with FITC conjugated anti-bovine polyclonal antibody.
Leucocyte subpopulations: Peripheral blood was collected in acid citrate dextrose (ACD). Mononuclear leucocytes were separated from other blood components by centrifugation on Lymphopaque (density: 1.086g/ml, sigma). Purified mononucleated leucocytes were reacted with a panel of monoclonal antibodies (MoAbs) specific to bovine leucocyte differentiation antigens. Fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG+M antibody (Caltag Lab., San Francisco) was used as a secondary antibody followed by reaction with MoAbs and separated cells. After fixing the cells on 96-well plates using 2% formaldehyde, proportions of leucocyte subpopulations were determined by using flow cytometry (Becton Dickinson, FACScan) and Consort 32 programs.
RESULTS
Therapeutic effects in calf respiratory disease: Table 1 summarises the clinical recovery rates (Excellent to Good) within 3 days for each treatment.
Table 1. Treatment group, clinical recovery rate (and % recovered) in calves with respiratory disease Treated in Group = Treatment Spring Summer Overall 1 = Medicine only 3/ 5 (60) 7/12 (58) 10/17 (59) 2 = Medicine + AP at SuQi 9/10 (90) 6/ 8 (75) 15/18 (83) 3 = Medicine + AP at (SanTai + FeiYu + AnFu) 4/ 5 (80) 7/10 (70) 11/15 (73) 4 = AP at SuQi only 13/15 (87) 6/ 8 (75) 19/23 (83) 5 = AP at (SanTai + FeiYu + AnFu) only 5/ 7 (71) 5/ 9 (55) 10/16 (63) Total 34/42 (81) 31/47 (66) 65/89 (73)
Table 1 shows that the curative effects of treatments 2 to 5 seemed to be 10-16% points better in spring than in summer but further research is needed to confirm this finding. Both combinations of medicine with AP (Groups 2 and 3, 83 and 73% respectively, overall 79%) gave higher recovery rates than medicine alone (59%). AP alone (Groups 4 and 5, 83 and 63% respectively, overall 74%) gave intermediate results. However, Groups 2 and 4 (Medicine + AP at SuQi and AP at SuQi alone) gave the best clinical results (83 and 83%).
Virus isolation and viral seroconversion rate: In calves treated by AP at SuQi, virus isolation fell from 77% before treatment to 54% on day 3 and 57% on day 10 after AP. The combination of medicine + SuQi showed similar results to those of AP alone (Table 2).
AP at SuQi gave 73% antibody production against BVD but gave lower antibody production (13%, and 27%) against IBR and PI3. However, the combination of medicine + AP at SuQi gave a lower antibody production than that of AP alone (Table 3).
Percentage of leucocyte subpopulation expressing specific antigen: Compared with pretreatment values, calves treated with AP, or medicine combined with AP, had:
a. significantly more leucocytes expressing MHC class II antigens (which have central roles in host defense mechanisms);
b. significantly more Bo (Bovine) CD2 and BoCD4 antigens and
c. a higher percentage of expression of B lymphocyte-specific antigen, surface IgM (sIgM), and N (nonT/nonB cells)-12 antigens (Table 4).
DISCUSSION
In our study of calves with respiratory disease, AP alone, or AP combined with medicine gave better clinical results than those of medicine alone. The improvement correlated with a decreased viral isolation rate, an increased number of negative samples and an increase in viral seroconversion rate. The therapeutic effects of AP may be due to the enhancement of host immunity by activation of lymphocytes involved in central host defense mechanisms.
The clinical recovery rate (Excellent to Good) of calves with respiratory disease in Group 1 (medicine alone) was 59%. Recovery rates were higher in Group 2 (medicine + AP at SuQi, 83%) and Group 4 (AP at SuQi alone, 83%). Recovery rates were intermediate in Group 3 (medicine + AP at (SanTai + FeiYu + AnFu), 73%) and Group 5 (AP at (SanTai + FeiYu + AnFu) alone, 63%).
AP alone decreased the rate of virus isolation in calves with respiratory disease and increased the number of negative samples. These effects were stronger for AP at SuQi than for AP at (SanTai + FeiYu + AnFu). We concluded that the changes of the rate of virus isolation and the number of negative samples may be the theoretical basis for the better therapeutic effects of SuQi relative to (SanTai + FeiYu + AnFu).
Although the overall clinical recovery rate to medicine + AP (79%) was slightly better than for AP alone (74%), the rate of virus isolation and the number of negative samples between the two groups were not correlated with the clinical results. Further study is needed to elucidate these results.
Viral seroconversion rate was low in calves treated by medicine + AP, whereas it was remarkably high in those treated by AP at SuQi. Compared with pre-treatment values, the percentage of leucocyte subpopulations expressing MHC class II antigens (which have central roles in host defence mechanism), CD2, CD4, B cells and N cells, were significantly higher in calves with respiratory disease treated with AP alone, or with medicine + AP. This may be the scientific basis for the therapeutic effects of AP alone or medicine + AP on SuQi in respiratory diseases in calves. Even though clinical recovery rates to the combined use of medicine + AP were somewhat better than those of AP alone, the isolation rate of virus and immune response showed no difference between those groups. This remains to be elucidated.
CONCLUSIONS
This trial used 89 calves with clinical respiratory disease to compare the therapeutic effects of medicine alone (injection of enrofloxacin, analgesic-antipyretics and antitussives), combinations of medicine + AP, and AP alone. AP was used for 3 days at one of two sets of points: SuQi alone, or a combination of (SanTai + AnFu + FeiYu).
Clinical recovery rates were poor (59%) in calves treated by medicine alone. Rates were best to combinations of medicine + AP (83-73%). AP alone gave intermediate rates (83-63%). AP alone at SuQi gave better recovery rates (83%) than at (SanTai + AnFu + FeiYu) (63%). Therapeutic effects of AP were related to enhanced host immune responses, by activating lymphocytes which express MHC (Major Histocompatibility Complex) class II, CD2 and CD4 antigens.
ACKNOWLEDGEMENTS
This paper was presented at the XIX World Buiatrics Congress, Edinburgh, 8-12 July, 1996. We thank Mr. Philip A.M. Rogers MRCVS, Dublin, Ireland for help in the post-Congress redrafting of the paper.
REFERENCES
1. Blood,D.C. & Radostits,O.M.: Veterinary Medicine 7th ed. Bailliere Tindall Toronto 1989, pp 367-372.
2. Berger,D. & Nolte,D.: Acupuncture in Bronchial Asthma. Comparative Medicine East and West 1977, 5: 265-269.
3. Da,Z.Z.: Treatment of Calf Bronchitis with TCM. J. Trad. Chin. Vet. Med. 1992, 4: 34-35.
4. Klide,A.M. & Kung,S.H.: Veterinary acupuncture. University of Pennsylvania Press 1977, pp 96-106.
5. Snow,D.H.: Pharmacological Basis of Large Animal Medicine. Eds J.A.Bogan, P.Lees, A.T.Yoxall. Blackwell Scientific Publication 1983, pp 391-427.
6. Selman,I.E.: Effects of Anti-prostaglandin Therapy in Experimental Parainfluenza type 3 Pneumonia in Weaned Conventional Calves. Vet. Rec. 1984, 109: 101-105.
7. Schwartz,C.: Acupuncture for Chronic Respiratory Conditions. In: Veterinary Acupuncture: Ancient Art to Modern Medicine (Ed: A.Schoen), American Veterinary Publications, 5782 Thornwood Drive, Goleta, CA93317, USA 1994, pp213-222.
8. Zwolfer,W., Keznickl-Hillebrand,W., Spacek,A., Cartellieri,M. & Grubofer,G.: Beneficial Effect of Acupuncture on Adult Patients with Bronchial Asthma. Am. J Chin. Med. 1993, 21: 113-117.
9. Yang,G.D. & Li,S.J.: Handbook of Veterinary Acupuncture, China Agriculture Press, Beijing 1985, pp 179-204.
10. Beijing Agricultural University, Chinese Veterinary Medicine, China Agriculture Press, Beijing 1986, pp 383-386.
11. Kurosawa,R. & Sakai,T.: Veterinary Surgery, Yang Hung Dang, Tokyo 1981, 569 pp.
12. Kim,Y.S. & Choe,Y.T.: Effect of Aqua-acupuncture of Sileris Radix Extract Solution on the Epithelium of the Trachea in Rats. Kyung Hee University, O. Med. J. 1984, 7: 335-344.
13. Song,Y.H. & Choe,Y.T.: Effects of On-chim, Laser acupuncture and acupuncture on the Depression of the Immune Response Induced by Cold stress in Mice. Kyung Hee University. O. Med. J. 1992, 15: 69-89.
14. Choo,T.C. & Choe,Y.T.: Effect of On-chim on Depression of Immune Response Induced by Exposed to Cold Stress in Mice. Kyung Hee University, O. Med. J. 1992, 15: 297-312.
15. Lin,J.H., Rogers,P.A.M. & Yamada,H.: The Scientific Basis of Chinese Herbal Medicine. In: Alternative Veterinary Medicine. Eds.: A.Schoen & S.Wynn, Mosby Book Publishers, USA 1996. In Press.
16. Rogers,P.A.M.: Immunologic Effects of Acupuncture. In: Veterinary Acupuncture: Ancient Art to Modern Medicine (Ed: A.Schoen), American Veterinary Publications, 5782 Thornwood Drive, Goleta, CA93317, USA 1994: pp243-267.
Table 2. Rate of virus isolation in calves with respiratory disease treated by AP at SuQi alone, or with medicine + AP at SuQi.
| Virus |
Days of treatment AP on SuQi alone Medicine + AP at SuQi |
|||||
| Day
1 n (%) |
Day
3 n (%) |
Day
10 n (%) |
Day
1 n (%) |
Day
3 n (%) |
Day
10 n (%) |
|
| Negative BVD + IBR BVD + IBR + PI3 IBR IBR + PI3 PI3 PI3 + BVD BVD |
4
(24) 4 (24) 2 (12) 1 ( 6) 2 (12) 1 ( 6) 2 (12) 1 ( 6) |
6
(46) 2 (15) 1 ( 8) 2 (15) 1 ( 8) 0 ( 0) 1 ( 8) 0 ( 0) |
6
(43) 2 (14) 1 ( 7) 2 (14) 0 ( 0) 0 ( 0) 0 ( 0) 3 (21) |
3
(30) 1 (10) 1 (10) 3 (30) 1 (10) 1 (10) 0 ( 0) 0 ( 0) |
4
(40) 2 (20) 1 (10) 3 (30) 0 ( 0) 0 ( 0) 0 ( 0) 0 ( 0) |
2
(20) 0 ( 0) 3 (30) 3 (30) 0 ( 0) 0 ( 0) 1 (10) 1 (10) |
Table 3. Effects on antibody production in calves with respiratory disease treated by AP at SuQi alone, or with medicine + AP at SuQi.
| AP on SuQi alone | Medicine + AP at SuQi | |||
| Virus |
Seroconversion n (%) |
Nonconversion n (%) |
Seroconversion n (%) |
Nonconversion n (%) |
| BVD IBR PI-3 |
11
(73) 2 (13) 4 (27) |
4
(27) 13 (86) 11 (73) |
2
(20) 0 ( 0) 1 (10) |
8 (
80) 10 (100) 9 ( 90) |
Table 4. Mean percentage and standard deviation (X%+sd) of leucocyte subpopulations expressing specific antigens in blood from calves with respiratory disease treated by AP at SuQi alone, or with medicine + AP at SuQi.
| AP on SuQi alone | Medicine + AP at SuQi | ||||||
| Antigens |
Monoclonal antibodies |
Before
AP (n=10) X%+sd |
Day
3 (n=10) X%+sd |
Day
10 (n=10) X%+sd |
Before AP (n=10) X%+sd |
Day
3 (n=10) X%+sd |
Day
10 (n=10) X%+sd |
| MHC
class I MHC class II BoCD 2 BoCD 4 BoCD 8 sIgM NI2 |
H58A H42A BAQ95A CACT138A CACT80C PIG45A CACT61A |
85.9+8.3 28.5+4.7 49.3+3.6 19.7+3.4 10.7+4.2 20.1+4.3 28.3+3.7 |
95.8+4.2 39.9+3.7 53.8+6.2 35.5+3.6 10.7+3.7 34.0+2.9 26.4+3.9 |
93.0+5.9 38.2+3.9 59.4+5.7 22.6+3.7 23.9+3.9 32.3+6.7 31.0+4.9 |
81.5+4.2 27.4+2.7 52.3+4.6 20.7+3.7 18.5+3.9 10.7+3.7 23.1+4.1 |
92.5+3.2 34.1+2.7 53.7+3.8 30.2+4.7 18.6+3.1 30.0+2.3 32.2+2.1 |
92.4+2.1 36.3+2.2 56.0+3.7 20.6+4.1 27.6+3.2 30.3+4.0 31.2+3.8 |