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The Effect of Antibacterials on Experimentally Induced Salmonella Typhimurium Infection in Chickens
307
B O N E STRENGTH
effect of wire pens, floor pens and cages on bone characteristics of laying hens. Poultry Sci. 49: 12231225. Rowland, L. O., Jr., J. L. Fry, R. B. Christmas, A. W. O'Steen and R. H. Harms, 1972. Differences in tibia strength and bone ash among strains of layers. Poultry Sci. 51: 1612-1615.
Snell, F. D., and L. S. Ettre, (Editors), 1969. Encyclopedia of Industrial Chemical Analysis, Vol. 9, p. 96. Interscience, John Wiley & Son, Inc., New York. Wabeck, C. J., and L. H. Littlefield, 1972. Bone strength of broilers reared in floor pens and cages having different bottoms. Poultry Sci. 51: 897-899.
MARGARET M . M A C K E N Z I E AND B . S. BAINS
Provincial Traders Pry. Limited, Momingside, Queensland 4170, Australia (Received for publication May 17, 1973)
ABSTRACT Chickens raised on wire floors were infected experimentally at 4 days of age with 8 x 10,0 cells of Salmonella typkimurium. Twenty four hours post-inoculation, infected chickens were treated with one of the following therapeutic substances—oxytetracycline, chlortetracycline, sulphadimidine, furaltadone, neomycin, furazolidone, chloramphenicl and ampicillin. Cloacal swabs were taken daily from each bird and cultured for Salmonella prior to autopsy examination on the 7th day when the visceral organs were subjected to bacteriological examination. Persistence of infection, mortality and average percent weight gains were correlated with the in vivo efficiency of the therapeutic substances. The results showed that chloramphenicol and furaltadone were the most efficient antibacterials for control of an experimental S. typhimunum infection in young chickens. POULTRY SCIENCE 53: 307-310, 1974
INTRODUCTION ALMONELLAE infect both man and animals with varying degrees of pathogenicity. Salmonellosis in poultry is not only of major economic importance, but also significant from the human health point of view. S. typhimunum is the predominant serotype occurring in both man and poultry. The shed pattern of chickens inoculated orally with S. typhimunum has been investigated by several workers. Milner and Shaffer (1952) inoculated day-old chicks orally with 1-5, 10, 102, 103, 104 and 105 cells of S. typhimunum, and correlated the faecal shedding pattern with inoculum size. They found that susceptibility to infection by the oral route was less at 4 days old than at 1 day old, and that infection at 1, 2, 3, 4 and 7 weeks old was detectable only with large
S
doses, and then only irregularly. Sadler et al. (1969) inoculated chickens at 2 days, 1, 2, 4 and 8 weeks with 102, 104, 106, 108 or 1010 viable cells of S. typhimurium, and studied the shed pattern by cloacal swabs, correlating the level of intestinal infection with bird age and inoculum dose. Fanelli et al. (1971) studied the shed pattern in 4 week chickens inoculated with 108 S. typhimunum cells, sampling by cloacal swabs on 0, 1, 2, 3, 6, 9, 13 and 17 days post-inoculation. A number of antibacterials have been widely used in the treatment of Salmonella infection in chickens, including oxytetracycline, chlortetracycline, sulphadimidine, furaltadone, neomycin, furazolidone, chloramphenicol and ampicillin. This study was made to determine the effect of treatment
Downloaded from http://ps.oxfordjournals.org/ at National Kaohsiung University of Applied Sciences on June 23, 2015
The Effect of Antibacterials on Experimentally Induced Salmonella Typhimunum Infection in Chickens
308
M. M. MACKENZIE AND B. S. BAINS
with these antibacterials on the shed pattern and subsequent isolation of S. typhimurium in chickens.
TABLE 1.—Antibacterial treatment of experimental groups Group
MATERIALS AND METHODS
1 2 3 4
Treatment Oxytetracycline Chlortetracycline Sulphadimidine Furaltadone
Dose and administration
Chicks from a commercial hatchery were placed on wire floors and brooded in isolated quarters to 11 days of age. Brooding temper5 Neomycin 6 Furazolidone ature was maintained at 37° C. and each group 7 Chloramphenicol was well isolated. Feed, continuously moni8 Ampicillin tored for Salmonella contamination, and water, were provided in excess. 9 Nontreated — 10 Noninfected — The test strain of S. typhimurium used was a recent isolate from a field outbreak of salmonellosis in broiler chickens. To determine its degree of pathogenicity in 4 day old Rectal swabs were examined daily from chicks, the LD50 dose was calculated; this all birds in groups 1-10 until autopsy on the was 16 x 10l0 organisms per chick. In an 7th day post-inoculation of Salmonella, when in vitro plate sensitivity test, growth of the heart, liver, spleen, intestine, caecum and culture was inhibited by furaltadone,1 fura- bone marrow were cultured for Salmonella. zolidone,2 chloramphenicol3 and ampicillin.4 RESULTS AND DISCUSSION The culture was resistant to oxytetracycline,5 6 7 chlortetracycline, sulphadimidine and All rectal swabs taken prior to inoculation neomycin.8 of Salmonella, and all samples of feed were At the start of the experiment, day-old birds negative for Salmonella. The results of the were divided into 10 groups of 11 each, each daily swabs following Salmonella inoculation bird being identified by a wingband. Rectal until autopsy on the 7th day are presented swabs were taken from each bird for 3 days in Figure 1. prior to inoculation with S. typhimurium; any The mortality and recovery rates of the birds found harbouring a natural Salmonella birds in each group are given in Table 2. infection were discarded. On the 4th day, The rectal swabs of all non-infected birds birds in groups 1-9 were inoculated per os (Group 10) remained negative throughout the with 8 x 1010 viable cells of 5. typhimurium. trial. All Salmonella -inoculated birds (Group The 10th group remained as non-infected 1-9) became Salmonella -carriers within 72 controls. hours of Salmonella dosage. Rectal swabs of An oral 5-day antibacterial treatment of the untreated birds (Group 9) remained posigroups 1-8 was commenced 24 hours later tive throughout the trial. Mortalities occurred in 4/11 birds in Group 9, compared with as shown in Table 1. none in the non-infected birds, Group 10. In the oxytetracycline-treated birds, Group 1, the high rate of shedding observed after 1,2. Watts Winter Pty. Ltd., Brisbane, Queensland, Australia. 72 hours was decreased to 70% by the 6th 3,5,6,7. Chemical Resources Pty. Ltd., Sydney, day. Mortalities occurred in 2 birds (18%) N.S.W., Australia. on the 2nd and 3rd days respectively. On 4. Beecham Laboratories, Sydney, N.S.W., autopsy examination, 6/9 birds were found Australia. to be still harbouring Salmonella: recovery 8. Upjohn Pty. Ltd., Sydney, N.S.W., Australia.
Downloaded from http://ps.oxfordjournals.org/ at National Kaohsiung University of Applied Sciences on June 23, 2015
25g./40 gals, of water 25g./40 gals, of water 3.5g./gal. of water 454g./160 gals, of water 200g./tonof feed 0.11% in feed 2 mg./day/chick in water 1 mg./day/chick in water
309
ANTIBACTERIALS AND S. TYPHIMURIUM
Gp.l. Oxytetracycline
100
Gp.2. Gp.3. Gp.4. Gp.5. Chlortetracycline Sulphadimidine Furaltadone Neomycin
75 50
1 2 3 4 5 6 7
Gp. 6. Furazolidone
Gp. 7. Chloramphenicol
1 2 3 4 5 6 7 Gp. S. Ampicillin
1 2 3 4 5 6 7 Gp. 9. Nontreated
1 2 3 4 5 6 7 Gp. 10. Noninfectsd
25 . I II I I I I I
FIG. 1. Daily and cumulative faecal shed of Salmonella typhimurium B5H Percent identified as shedding on a given day f w 3 Percent having been identified at least once as shedding I I None identified as shedding (a) Days post-inoculation with Salmonella typhimurium TABLE 2.—Mortality and recovery rates of groups 1-10. Group no. 1 2 3 4 5 6 7 8 9 10
Sal. typhimurium inoculation
+ +
+ + + + +
+ + —
Treatment Oxytetracycline Chlortetracycline Sulphadimidine Furasol Neomycin Neftin Chloramphenicol Ampicillin
— —
from infection was observed in only 3/11 birds. Similarly, in Groups 2, 3, 5 and 6 representing the chlortetracycline-, sulphadimidine-, neomycin- and furazolidone-treated birds, treatment did not greatly decrease the proportion of carriers in a group (2/11 in all groups) despite the fact that in vitro the culture had been sensitive to furazolidone. Mortalities in these groups were 1/11, 4 / 1 1 ,
Mortalities 2/11 1/11 4/11 1/11 2/11 1/11 1/11 3/11 4/11 0/11
No. recovered from infection on autopsy 3/11 2/11 2/11 6/11 2/11 2/11 8/11 5/11 0/11
—
2/11 and 1/11 respectively, over the 7-day period. By contrast, treatment of Groups 4, 7 and 8 with furaltadone, chloramphenicol and ampicillin respectively, resulted in a significant decrease in the shedding rate over the 7-day period. Most efficient treatment was obtained with chloramphenicol which resulted in early (4th day) suppression of Salmonella shedding to a basal level of 20%.
Downloaded from http://ps.oxfordjournals.org/ at National Kaohsiung University of Applied Sciences on June 23, 2015
(a)l £ 3 4 5 6 7
310
M. M. MACKENZIE AND B. S. BAINS
TABLE 3.—Percent weight gains of groups 1-10 Av. % weight gains 130 130 131 148 123 123 138 108 108 135
Autopsy revealed 72% of the birds in this group to be negative in culturing for Salmonella, although the drug has bacteriostatic rather than bacteriocidal properties. Furaltadone produced a slower decline in shedding rate, decreasing to a basal level of 40% on the 6th day; 60% of birds were Salmonella negative on the 7th day. Mortality of both the chloramphenicol- and furaltadone- groups was 1/11 (9%). Although a comparatively high rate of recovery (50%) was obtained in the ampicillin-treated birds (Group 8) mortalities of 27% occurred. The average percentage weight gains obtained for the birds in each group over the 7-day period are shown in Table 3. Some stunting occurred in the Salmonellainoculated, untreated controls. No significant differences were observed in percentage weight gains on the antibiotic treatments in
ACKNOWLEDGMENT The authors wish to express their gratitude to Mrs. H. Adam for able technical assistance through this work. REFERENCES Fanelli, M. J., W. W. Sadler, C. E. Franti and J. R. Brownell, 1971. Localization of Salmonellae within the intestinal tract of chickens. Av. Dis. 15: 366-375. Milner, K. C , and M. F. Shaffer, 1952. Bacteriologic studies of experimental Salmonella infections in chicks. J. Infect. Dis. 90: 81. Sadler, W. W., J. R. Brownell and M. J. Fanelli, 1969. Influence of age and inoculum level on shed pattern of Salmonella typhimurium in chickens. Av. Dis. 13: 793.
NEWS AND NOTES (Continued from page 272) D.C. office. He is International Director for Market Development. R. J. Andrews has been appointed International Director for Market Development. He will assume U.S. supervision of I.T.D. programs. Lee Campbell, who has been handling this task, will have more time for increased attention to government relations work.
ARKANSAS NOTES A $5,000 grant from Midwest Solvents Co., Inc., Atchison, Kansas, has been given to the University of Arkansas Division of Agriculture. An important source of high protein feed in the United States comes from distilleries and industries
(Continued on page 348)
Downloaded from http://ps.oxfordjournals.org/ at National Kaohsiung University of Applied Sciences on June 23, 2015
Group no. Treatment 1 Oxytetracycline 2 Chlortetracycline 3 Sulphadimidine 4 Furasol 5 Neomycin 6 Neftin 7 Chloramphenicol 8 Ampicillin 9 Salmonella-untreated controls 10 Normal controls
Groups 1-7. The ampicillin-treated group, however, exhibited similar weight gains to the untreated Salmonella controls. The findings indicate that the most efficient drugs for treatment of S. typhimurium infection in chickens are chloramphenicol and furaltadone. Although ampicillin allows relatively high recoveries from infection, some stunting of the birds and higher mortalities do occur. Considerably lower rates of recovery from infection are obtained with the remaining antibacterials, although mortalities are reduced compared with the untreated Salmonella controls, except in the case of sulphadimidine where mortality is the same as in the controls.
B O N E STRENGTH
effect of wire pens, floor pens and cages on bone characteristics of laying hens. Poultry Sci. 49: 12231225. Rowland, L. O., Jr., J. L. Fry, R. B. Christmas, A. W. O'Steen and R. H. Harms, 1972. Differences in tibia strength and bone ash among strains of layers. Poultry Sci. 51: 1612-1615.
Snell, F. D., and L. S. Ettre, (Editors), 1969. Encyclopedia of Industrial Chemical Analysis, Vol. 9, p. 96. Interscience, John Wiley & Son, Inc., New York. Wabeck, C. J., and L. H. Littlefield, 1972. Bone strength of broilers reared in floor pens and cages having different bottoms. Poultry Sci. 51: 897-899.
MARGARET M . M A C K E N Z I E AND B . S. BAINS
Provincial Traders Pry. Limited, Momingside, Queensland 4170, Australia (Received for publication May 17, 1973)
ABSTRACT Chickens raised on wire floors were infected experimentally at 4 days of age with 8 x 10,0 cells of Salmonella typkimurium. Twenty four hours post-inoculation, infected chickens were treated with one of the following therapeutic substances—oxytetracycline, chlortetracycline, sulphadimidine, furaltadone, neomycin, furazolidone, chloramphenicl and ampicillin. Cloacal swabs were taken daily from each bird and cultured for Salmonella prior to autopsy examination on the 7th day when the visceral organs were subjected to bacteriological examination. Persistence of infection, mortality and average percent weight gains were correlated with the in vivo efficiency of the therapeutic substances. The results showed that chloramphenicol and furaltadone were the most efficient antibacterials for control of an experimental S. typhimunum infection in young chickens. POULTRY SCIENCE 53: 307-310, 1974
INTRODUCTION ALMONELLAE infect both man and animals with varying degrees of pathogenicity. Salmonellosis in poultry is not only of major economic importance, but also significant from the human health point of view. S. typhimunum is the predominant serotype occurring in both man and poultry. The shed pattern of chickens inoculated orally with S. typhimunum has been investigated by several workers. Milner and Shaffer (1952) inoculated day-old chicks orally with 1-5, 10, 102, 103, 104 and 105 cells of S. typhimunum, and correlated the faecal shedding pattern with inoculum size. They found that susceptibility to infection by the oral route was less at 4 days old than at 1 day old, and that infection at 1, 2, 3, 4 and 7 weeks old was detectable only with large
S
doses, and then only irregularly. Sadler et al. (1969) inoculated chickens at 2 days, 1, 2, 4 and 8 weeks with 102, 104, 106, 108 or 1010 viable cells of S. typhimurium, and studied the shed pattern by cloacal swabs, correlating the level of intestinal infection with bird age and inoculum dose. Fanelli et al. (1971) studied the shed pattern in 4 week chickens inoculated with 108 S. typhimunum cells, sampling by cloacal swabs on 0, 1, 2, 3, 6, 9, 13 and 17 days post-inoculation. A number of antibacterials have been widely used in the treatment of Salmonella infection in chickens, including oxytetracycline, chlortetracycline, sulphadimidine, furaltadone, neomycin, furazolidone, chloramphenicol and ampicillin. This study was made to determine the effect of treatment
Downloaded from http://ps.oxfordjournals.org/ at National Kaohsiung University of Applied Sciences on June 23, 2015
The Effect of Antibacterials on Experimentally Induced Salmonella Typhimunum Infection in Chickens
308
M. M. MACKENZIE AND B. S. BAINS
with these antibacterials on the shed pattern and subsequent isolation of S. typhimurium in chickens.
TABLE 1.—Antibacterial treatment of experimental groups Group
MATERIALS AND METHODS
1 2 3 4
Treatment Oxytetracycline Chlortetracycline Sulphadimidine Furaltadone
Dose and administration
Chicks from a commercial hatchery were placed on wire floors and brooded in isolated quarters to 11 days of age. Brooding temper5 Neomycin 6 Furazolidone ature was maintained at 37° C. and each group 7 Chloramphenicol was well isolated. Feed, continuously moni8 Ampicillin tored for Salmonella contamination, and water, were provided in excess. 9 Nontreated — 10 Noninfected — The test strain of S. typhimurium used was a recent isolate from a field outbreak of salmonellosis in broiler chickens. To determine its degree of pathogenicity in 4 day old Rectal swabs were examined daily from chicks, the LD50 dose was calculated; this all birds in groups 1-10 until autopsy on the was 16 x 10l0 organisms per chick. In an 7th day post-inoculation of Salmonella, when in vitro plate sensitivity test, growth of the heart, liver, spleen, intestine, caecum and culture was inhibited by furaltadone,1 fura- bone marrow were cultured for Salmonella. zolidone,2 chloramphenicol3 and ampicillin.4 RESULTS AND DISCUSSION The culture was resistant to oxytetracycline,5 6 7 chlortetracycline, sulphadimidine and All rectal swabs taken prior to inoculation neomycin.8 of Salmonella, and all samples of feed were At the start of the experiment, day-old birds negative for Salmonella. The results of the were divided into 10 groups of 11 each, each daily swabs following Salmonella inoculation bird being identified by a wingband. Rectal until autopsy on the 7th day are presented swabs were taken from each bird for 3 days in Figure 1. prior to inoculation with S. typhimurium; any The mortality and recovery rates of the birds found harbouring a natural Salmonella birds in each group are given in Table 2. infection were discarded. On the 4th day, The rectal swabs of all non-infected birds birds in groups 1-9 were inoculated per os (Group 10) remained negative throughout the with 8 x 1010 viable cells of 5. typhimurium. trial. All Salmonella -inoculated birds (Group The 10th group remained as non-infected 1-9) became Salmonella -carriers within 72 controls. hours of Salmonella dosage. Rectal swabs of An oral 5-day antibacterial treatment of the untreated birds (Group 9) remained posigroups 1-8 was commenced 24 hours later tive throughout the trial. Mortalities occurred in 4/11 birds in Group 9, compared with as shown in Table 1. none in the non-infected birds, Group 10. In the oxytetracycline-treated birds, Group 1, the high rate of shedding observed after 1,2. Watts Winter Pty. Ltd., Brisbane, Queensland, Australia. 72 hours was decreased to 70% by the 6th 3,5,6,7. Chemical Resources Pty. Ltd., Sydney, day. Mortalities occurred in 2 birds (18%) N.S.W., Australia. on the 2nd and 3rd days respectively. On 4. Beecham Laboratories, Sydney, N.S.W., autopsy examination, 6/9 birds were found Australia. to be still harbouring Salmonella: recovery 8. Upjohn Pty. Ltd., Sydney, N.S.W., Australia.
Downloaded from http://ps.oxfordjournals.org/ at National Kaohsiung University of Applied Sciences on June 23, 2015
25g./40 gals, of water 25g./40 gals, of water 3.5g./gal. of water 454g./160 gals, of water 200g./tonof feed 0.11% in feed 2 mg./day/chick in water 1 mg./day/chick in water
309
ANTIBACTERIALS AND S. TYPHIMURIUM
Gp.l. Oxytetracycline
100
Gp.2. Gp.3. Gp.4. Gp.5. Chlortetracycline Sulphadimidine Furaltadone Neomycin
75 50
1 2 3 4 5 6 7
Gp. 6. Furazolidone
Gp. 7. Chloramphenicol
1 2 3 4 5 6 7 Gp. S. Ampicillin
1 2 3 4 5 6 7 Gp. 9. Nontreated
1 2 3 4 5 6 7 Gp. 10. Noninfectsd
25 . I II I I I I I
FIG. 1. Daily and cumulative faecal shed of Salmonella typhimurium B5H Percent identified as shedding on a given day f w 3 Percent having been identified at least once as shedding I I None identified as shedding (a) Days post-inoculation with Salmonella typhimurium TABLE 2.—Mortality and recovery rates of groups 1-10. Group no. 1 2 3 4 5 6 7 8 9 10
Sal. typhimurium inoculation
+ +
+ + + + +
+ + —
Treatment Oxytetracycline Chlortetracycline Sulphadimidine Furasol Neomycin Neftin Chloramphenicol Ampicillin
— —
from infection was observed in only 3/11 birds. Similarly, in Groups 2, 3, 5 and 6 representing the chlortetracycline-, sulphadimidine-, neomycin- and furazolidone-treated birds, treatment did not greatly decrease the proportion of carriers in a group (2/11 in all groups) despite the fact that in vitro the culture had been sensitive to furazolidone. Mortalities in these groups were 1/11, 4 / 1 1 ,
Mortalities 2/11 1/11 4/11 1/11 2/11 1/11 1/11 3/11 4/11 0/11
No. recovered from infection on autopsy 3/11 2/11 2/11 6/11 2/11 2/11 8/11 5/11 0/11
—
2/11 and 1/11 respectively, over the 7-day period. By contrast, treatment of Groups 4, 7 and 8 with furaltadone, chloramphenicol and ampicillin respectively, resulted in a significant decrease in the shedding rate over the 7-day period. Most efficient treatment was obtained with chloramphenicol which resulted in early (4th day) suppression of Salmonella shedding to a basal level of 20%.
Downloaded from http://ps.oxfordjournals.org/ at National Kaohsiung University of Applied Sciences on June 23, 2015
(a)l £ 3 4 5 6 7
310
M. M. MACKENZIE AND B. S. BAINS
TABLE 3.—Percent weight gains of groups 1-10 Av. % weight gains 130 130 131 148 123 123 138 108 108 135
Autopsy revealed 72% of the birds in this group to be negative in culturing for Salmonella, although the drug has bacteriostatic rather than bacteriocidal properties. Furaltadone produced a slower decline in shedding rate, decreasing to a basal level of 40% on the 6th day; 60% of birds were Salmonella negative on the 7th day. Mortality of both the chloramphenicol- and furaltadone- groups was 1/11 (9%). Although a comparatively high rate of recovery (50%) was obtained in the ampicillin-treated birds (Group 8) mortalities of 27% occurred. The average percentage weight gains obtained for the birds in each group over the 7-day period are shown in Table 3. Some stunting occurred in the Salmonellainoculated, untreated controls. No significant differences were observed in percentage weight gains on the antibiotic treatments in
ACKNOWLEDGMENT The authors wish to express their gratitude to Mrs. H. Adam for able technical assistance through this work. REFERENCES Fanelli, M. J., W. W. Sadler, C. E. Franti and J. R. Brownell, 1971. Localization of Salmonellae within the intestinal tract of chickens. Av. Dis. 15: 366-375. Milner, K. C , and M. F. Shaffer, 1952. Bacteriologic studies of experimental Salmonella infections in chicks. J. Infect. Dis. 90: 81. Sadler, W. W., J. R. Brownell and M. J. Fanelli, 1969. Influence of age and inoculum level on shed pattern of Salmonella typhimurium in chickens. Av. Dis. 13: 793.
NEWS AND NOTES (Continued from page 272) D.C. office. He is International Director for Market Development. R. J. Andrews has been appointed International Director for Market Development. He will assume U.S. supervision of I.T.D. programs. Lee Campbell, who has been handling this task, will have more time for increased attention to government relations work.
ARKANSAS NOTES A $5,000 grant from Midwest Solvents Co., Inc., Atchison, Kansas, has been given to the University of Arkansas Division of Agriculture. An important source of high protein feed in the United States comes from distilleries and industries
(Continued on page 348)
Downloaded from http://ps.oxfordjournals.org/ at National Kaohsiung University of Applied Sciences on June 23, 2015
Group no. Treatment 1 Oxytetracycline 2 Chlortetracycline 3 Sulphadimidine 4 Furasol 5 Neomycin 6 Neftin 7 Chloramphenicol 8 Ampicillin 9 Salmonella-untreated controls 10 Normal controls
Groups 1-7. The ampicillin-treated group, however, exhibited similar weight gains to the untreated Salmonella controls. The findings indicate that the most efficient drugs for treatment of S. typhimurium infection in chickens are chloramphenicol and furaltadone. Although ampicillin allows relatively high recoveries from infection, some stunting of the birds and higher mortalities do occur. Considerably lower rates of recovery from infection are obtained with the remaining antibacterials, although mortalities are reduced compared with the untreated Salmonella controls, except in the case of sulphadimidine where mortality is the same as in the controls.