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Research Note: Anticoccidial Effect of Monensin Against Eimeria mitis and Eimeria dispersa
Research Note: Anticoccidial Effect of Monensin Against Eimeria mitis and Eimeria dispersa KEVIN L. WATKINS, KENNETH W. BAFUNDO, and DAVID J. DONOVAN Lilly Research Laboratories, A Division of Eli Lilly and Company, P.O. Box 708, Greenfield, Indiana 46140 (Received for publication November 13, 1989)
1990 Poultry Science 69:1009-1011
investigate the effect of monensin on experimentally produced infections of Eimeria mitis In 1929, Tyzzer characterized both Eimeria and Eimeria dispersa in chicks and poults, mitis in chickens and Eimeria dispersa in respectively. turkeys. Because of their relatively slight pathogenicity, these two protozoans usually are MATERIALS AND METHODS not considered to be of significant economic importance for the poultry industry. Although Male, Hubbard chicks were fed a basal early investigations noted the lack of gross ration wim 23.5% erode protein from 0 to 7 pathological changes and lesions of the intes- days after hatching. Male, Nicholas poults tine, many studies have demonstrated a reduc- were fed a basal ration with 30% crude protein tion in growth, pigmentation, and absorptive from 0 to 12 days after hatching (Table 1). All capacity among birds infected with E. mitis birds were obtained from commercial hatcher(Joyner, 1958; Ruff and Edgar, 1982; Allen, ies. On Days 7 and 12 after hatching, the 1987) and with E. dispersa (Oluleye and respective chicks and poults were randomly Edgar, 1980; Ruff et al, 1981); also, marked assigned to treatment group on basis of body microscopic signs of intestinal damage have weight. All birds were raised in stainless-steel been reported with such infections (Madden batteries. Feed, water, and lighting were and Ruff, 1979; Ruff et al, 1981; Novilla et provided continuously. al., 1987). Four chick trials were conducted using 13 Although monensin will control many spe- replicates each for the uninfected, unmedicated cies of Eimeria in both chickens (E. acer- chicks and for the infected, unmedicated vulina, E. maxima, E. tenella, E. brunetti, E.chicks. There were 17 replicates for the mivati, and E. necatrix) and in turkeys (E. infected chicks fed the basal containing 100 gallopavonis, E. meleagrimitis, and E. ppm of monensin. Five poult trials were adenoeides), little or no data are available conducted using a total of 20 replicates each of demonstrating the anticoccidial activity of uninfected, unmedicated poults; infected, unmonensin against E. mitis and E. dispersa. medicated poults; and of infected poults fed Therefore, several studies were conducted to the basal containing 60 ppm of monensin. INTRODUCTION
1009
Downloaded from http://ps.oxfordjournals.org/ at Rutgers University Libraries/Technical Services on June 1, 2015
ABSTRACT Four chick and five poult trials were conducted in order to investigate the anticoccidial efficacy of monensin against Eimeria mitis in chickens and Eimeria dispersa in turkeys. The chicks were fed a basal diet with either 0 or 100 ppm of monensin. The poults were fed a basal diet with either 0 or 60 ppm of monensin. Two days after the initiation of each experiment, the chicks and poults were crop-intubated with oocysts of E. mitis and E. dispersa, respectively. A group was also included that was not infected and not medicated. Growth and feed intake were recorded. At 6 or 7 days postinoculation, the birds were killed by cervical dislocation and were scored for the incidence and severity of intestinal abnormalities. The Eimeria mitis infection reduced (P<01) gain and the feed:gain ratio, compared with uninfected birds; the E. dispersa infection only reduced (P<05) gain. Although well-defined, discrete lesions were not observed, marked intestinal abnormalities were noted in birds infected with either E. mitis or E. dispersa. Also, the infected, unmedicated birds had increased (P<01) intestinal scores compared with uninfected birds or those with infection but treated with monensin. Monensin eliminated the reduction in gain and feed efficiency seen in the infected birds. The infected birds fed monensin had intestinal scores, gain, and feed:gain ratios similar (P>. 10) to those for the birds that were not infected and that did not receive medication. (Key words: growth, intestinal scores, coccidiosis, monensin, Eimeria mitis, Eimeria dispersa)
1010
WATCONS ET AL. TABLE 1. Dietary composition 1
Chick
Ground com Soybean meal (48%) Distillers dried solubles Fish meal Animal fat Alfalfa leaf meal Dicalcium phosphate Limestone Salt Vitamin premix3 Trace-mineral premix4 Methionine hydroxy analog Selenium premix5
53.42 31.73 4.00 5.00 2.83
POult
(%)
1.28 .62 .30 .50 .10 .17 .05
31.85 46.00 5.00 5.00 5.00 2.50 2.20 1.40 .30 .50 .10 .10 .05
Calculated composition: 23.5% CP, .564% methionine, .85% Ca, .5% P, and 3,100 kcal of ME per kg. Calculated composition: 30.0% CP, .62% methionine, 1.41% Ca, 1.0% P, and 2,907 kcal of ME per kg. 3 Vitamin premix provided per kilogram of diet: vitamin A, 3,000 IU; vitamin D 3 , 900 ICU; vitamin E, 40 mg; vitamin K, .7 mg; choline, 1,000 mg; niacin, 70 mg; pantothenic acid, 4 mg; riboflavin, 4 mg; vitamin Bj2, .10 mg; biotin, .10 mg; ethoxyquin, 125 mg. ^Trace-mineral premix provided per kilogram of diet: manganese, 75 mg; zinc, 50 mg; iron, 25 mg; iodine, 1 mg. Selenium premix provides .10 mg Se per kg of diet.
Each replicate consisted of either 4 or 5 birds per pen. The infected chicks were inoculated via crop-intubation with 250,000 sporulated E. mitis (F.S. 271) oocysts 48 h after the experiment began. The poults were inoculated in a similar manner with 20,000 E. dispersa (F.S. 647) oocysts. All birds-including the uninfected, unmedicated group-were scored for intestinal abnormalities. The chicks were scored on Day 7 after inoculation; the poults, on Day 6 after inoculation. The response criteria were body weight gain, feed:gain ratio, mortality, and intestinal scores. The intestines of the chicks and of the poults were removed. Three sites (anterior, mid, and posterior) of the small intestine were assigned a score from 0 to 4. Although discrete, intestinal lesions are not usually seen in either E. dispersa or E. mitis infections, other pathological signs are present. Visual intestinal scoring was based on epithelial color, fluid accumulation, and the overall general appearance of the intestine (serosal thickness, mucosal erosion, dilation, and similar factors).
An intestinal score of 0 was given to areas of the intestine that looked normal. Scores from 1 to 4 were used to rank the incidence and severity of the symptoms just described. The scores from each of the three intestinal sites were summed for analysis. The data were analyzed using analysis of variance procedures appropriate for a completely randomized design (Steel and Torrie, 1980). Since no interactions for treatment by trial were present, all data were pooled across trials. Due to heterogeneity of variance, the intestinal scores were transformed [log(+l)] prior to statistical analysis. Treatment differences were separated by the F-protected, least significant difference procedure. Pen was used as the experimental unit. RESULTS AND DISCUSSION
The results demonstrated that both E. mitis and E. dispersa infections can reduce growth as well as feed efficiency (Table 2). Although the species studied historically have been considered some of the least pathogenic, the present findings support those by others who reported that infections of E. mitis and E. dispersa can result in reduced growth and absorptive capacity (Joyner, 1958; Oluleye and Edgar, 1980; Ruff and Edgar, 1982; Allen, 1987). In the current study, Eimeria mitis and E. dispersa infections reduced gain 21 and 7% and reduced feed efficiency 14 and 4%. Eimeria mitis infection reduced (P<01) gain and the feed.gain ratio compared with uninfected birds, while E. dispersa infection only reduced (P<.05) gain. The lack of well-defined, discrete lesions in infections of E. mitis (Ruff and Edgar, 1982), E, dispersa (Madden and Ruff, 1979), and turkey coccidiosis in general (Jeffers and Bentley, 1980) made lesion scoring very difficult. Although gross intestinal lesions were not present in either E. mitis or E. dispersa infection, the authors did observe a reduced pigmentation, fluid accumulation, and a generalized abnormal appearance, resulting in relatively high intestinal scores (Table 2). Although the intestinal scores for birds infected with E. dispersa were similar to those for birds infected with E. mitis, the reduction in growth and feed efficiency due to E. dispersa infection was much less. No mortality occurred in any of the trials, although in some cases severe intestinal damage was noted.
Downloaded from http://ps.oxfordjournals.org/ at Rutgers University Libraries/Technical Services on June 1, 2015
Ingredient
2
1011
RESEARCH NOTE
TABLE 2. Effect of monensin on the growth and intestinal scores of chicks infected with Eimeria mitis and of poults infected with Eimeria dispersa Chicks infected with E. mitis
.O
Intestinal score
Gain
Feed:gain ratio
.00b 6.54" l-52 b
(£) 205 c 190d 205°
(g/g) 1.645 1.708 1.667
.76
3.79
.019
Intestinal score2 .51" 6.09" .44 b .51
"•"Means within columns with no common superscripts are significantly different (P<01). c,d Means within columns with no common superscripts are significantly different (P<.05). 1 NN=not infected, not medicated; IN=infected, not medicated; MON=infected, monensin-medicated. The chicks were fed diets containing 100 ppm of monensin; the poults were fed diets containing 60 ppm of monensin. ^Total score for the three intestinal sites, with a score of 12 being the highest possible. The intestinal scores were retransformed for table presentation. 3 Pooled.
Monensin was very effective in controlling the infections of both E. mitis and E. dispersa. The infected birds fed diets containing monensin had gains and feed:gain ratios similar (P>.10) to those of the uninfected birds. Monensin greatly reduced the incidence and severity of the intestinal abnormalities observed in the infected birds. No difference (P>.10) in intestinal scores was detected between the infected birds fed monensin and the uninfected, unmedicated birds. Oluleye and Edgar (1980) also reported that monensin was effective in controlling E. dispersa infections in turkeys. In summary, although E. mitis and E. dispersa are considered to be some of the lesspathogenic species of chicken and turkey coccidia, infection with either one can result in economic losses due to reductions in both growth and in feed efficiency. Monensin, when fed at 100 ppm to chicks or at 60 ppm to poults, provides excellent control of both E. mitis and E. dispersa. REFERENCES Allen, P. C , 1987. Physiological responses of chicken gut tissue to coccidial infection: Comparative effects of
Eimeria acervulina and Eimeria mitis on mucosal mass, carotenoid content, and brush border enzyme activity. Poultry Sci. 66:1306-1315. Jeffers, T. K., and E. J. Bentley, 1980. Monensin sensitivity of recent field isolates of turkey coccidia. Poultry Sci. 59:1722-1730. Joyner, L. P., 1958. Experimental fiwieria mitis infections in chickens. Parasitology 48:101-112. Madden, P. A., and M. D. Ruff, 1979. Eimeria dispersa, E. adenoeides, and E. meleagrimitis: intestinal mucosal disruption in turkeys as seen with scanning electron microscopy. J. Parasitol. 65:234-242. Novilla, M N., T. K. Jeffers, W. J. Griffing, and S. L. White, 1987. A redescription of the life cycle of Eimeria mitis, Tyzzer, 1929. J. Protozool. 34:87-92. Oluleye, O. R., and S. A. Edgar, 1980. Details of the life history and pathogenicity of the turkey coccidinm, Eimeria dispersa, Tyzzer 1929, and efficacy of four drugs for the control of the disease. Poultry Sci. 59(Suppl. 1):1569. (Abstr.) Ruff, M D., P. C. Augustine, and P. A. Madden, 1981. Eimeria meleagrimitis, E. adenoeides, and E. dispersa: severity of infection and changes in the intestinal mucosa of the turkey. Exp. Parasitol. 51:87-94. Ruff, M. D., and S. A. Edgar, 1982. Reduced intestinal absorption in broilers during Eimeria mitis infection. Am. J. Vet Res. 43:507-509. Steel, R.GX)., and J. H. Torrie, 1980. Principles and Procedures of Statistics: A Biometrical Approach. McGrawHill, Inc., New York, NY. Tyzzer, E. E., 1929. Coccidiosis in gallinaceous birds. Am. J. Hyg. 10:269-383.
Downloaded from http://ps.oxfordjournals.org/ at Rutgers University Libraries/Technical Services on June 1, 2015
c*i
(g) 288" 227 b 279" 9.38
at
NN IN MON SEM3
Feed:gain ratio X>
Gain
bo oo •£ 0\
Treatment1
Poults infected with E. dispersa
1990 Poultry Science 69:1009-1011
investigate the effect of monensin on experimentally produced infections of Eimeria mitis In 1929, Tyzzer characterized both Eimeria and Eimeria dispersa in chicks and poults, mitis in chickens and Eimeria dispersa in respectively. turkeys. Because of their relatively slight pathogenicity, these two protozoans usually are MATERIALS AND METHODS not considered to be of significant economic importance for the poultry industry. Although Male, Hubbard chicks were fed a basal early investigations noted the lack of gross ration wim 23.5% erode protein from 0 to 7 pathological changes and lesions of the intes- days after hatching. Male, Nicholas poults tine, many studies have demonstrated a reduc- were fed a basal ration with 30% crude protein tion in growth, pigmentation, and absorptive from 0 to 12 days after hatching (Table 1). All capacity among birds infected with E. mitis birds were obtained from commercial hatcher(Joyner, 1958; Ruff and Edgar, 1982; Allen, ies. On Days 7 and 12 after hatching, the 1987) and with E. dispersa (Oluleye and respective chicks and poults were randomly Edgar, 1980; Ruff et al, 1981); also, marked assigned to treatment group on basis of body microscopic signs of intestinal damage have weight. All birds were raised in stainless-steel been reported with such infections (Madden batteries. Feed, water, and lighting were and Ruff, 1979; Ruff et al, 1981; Novilla et provided continuously. al., 1987). Four chick trials were conducted using 13 Although monensin will control many spe- replicates each for the uninfected, unmedicated cies of Eimeria in both chickens (E. acer- chicks and for the infected, unmedicated vulina, E. maxima, E. tenella, E. brunetti, E.chicks. There were 17 replicates for the mivati, and E. necatrix) and in turkeys (E. infected chicks fed the basal containing 100 gallopavonis, E. meleagrimitis, and E. ppm of monensin. Five poult trials were adenoeides), little or no data are available conducted using a total of 20 replicates each of demonstrating the anticoccidial activity of uninfected, unmedicated poults; infected, unmonensin against E. mitis and E. dispersa. medicated poults; and of infected poults fed Therefore, several studies were conducted to the basal containing 60 ppm of monensin. INTRODUCTION
1009
Downloaded from http://ps.oxfordjournals.org/ at Rutgers University Libraries/Technical Services on June 1, 2015
ABSTRACT Four chick and five poult trials were conducted in order to investigate the anticoccidial efficacy of monensin against Eimeria mitis in chickens and Eimeria dispersa in turkeys. The chicks were fed a basal diet with either 0 or 100 ppm of monensin. The poults were fed a basal diet with either 0 or 60 ppm of monensin. Two days after the initiation of each experiment, the chicks and poults were crop-intubated with oocysts of E. mitis and E. dispersa, respectively. A group was also included that was not infected and not medicated. Growth and feed intake were recorded. At 6 or 7 days postinoculation, the birds were killed by cervical dislocation and were scored for the incidence and severity of intestinal abnormalities. The Eimeria mitis infection reduced (P<01) gain and the feed:gain ratio, compared with uninfected birds; the E. dispersa infection only reduced (P<05) gain. Although well-defined, discrete lesions were not observed, marked intestinal abnormalities were noted in birds infected with either E. mitis or E. dispersa. Also, the infected, unmedicated birds had increased (P<01) intestinal scores compared with uninfected birds or those with infection but treated with monensin. Monensin eliminated the reduction in gain and feed efficiency seen in the infected birds. The infected birds fed monensin had intestinal scores, gain, and feed:gain ratios similar (P>. 10) to those for the birds that were not infected and that did not receive medication. (Key words: growth, intestinal scores, coccidiosis, monensin, Eimeria mitis, Eimeria dispersa)
1010
WATCONS ET AL. TABLE 1. Dietary composition 1
Chick
Ground com Soybean meal (48%) Distillers dried solubles Fish meal Animal fat Alfalfa leaf meal Dicalcium phosphate Limestone Salt Vitamin premix3 Trace-mineral premix4 Methionine hydroxy analog Selenium premix5
53.42 31.73 4.00 5.00 2.83
POult
(%)
1.28 .62 .30 .50 .10 .17 .05
31.85 46.00 5.00 5.00 5.00 2.50 2.20 1.40 .30 .50 .10 .10 .05
Calculated composition: 23.5% CP, .564% methionine, .85% Ca, .5% P, and 3,100 kcal of ME per kg. Calculated composition: 30.0% CP, .62% methionine, 1.41% Ca, 1.0% P, and 2,907 kcal of ME per kg. 3 Vitamin premix provided per kilogram of diet: vitamin A, 3,000 IU; vitamin D 3 , 900 ICU; vitamin E, 40 mg; vitamin K, .7 mg; choline, 1,000 mg; niacin, 70 mg; pantothenic acid, 4 mg; riboflavin, 4 mg; vitamin Bj2, .10 mg; biotin, .10 mg; ethoxyquin, 125 mg. ^Trace-mineral premix provided per kilogram of diet: manganese, 75 mg; zinc, 50 mg; iron, 25 mg; iodine, 1 mg. Selenium premix provides .10 mg Se per kg of diet.
Each replicate consisted of either 4 or 5 birds per pen. The infected chicks were inoculated via crop-intubation with 250,000 sporulated E. mitis (F.S. 271) oocysts 48 h after the experiment began. The poults were inoculated in a similar manner with 20,000 E. dispersa (F.S. 647) oocysts. All birds-including the uninfected, unmedicated group-were scored for intestinal abnormalities. The chicks were scored on Day 7 after inoculation; the poults, on Day 6 after inoculation. The response criteria were body weight gain, feed:gain ratio, mortality, and intestinal scores. The intestines of the chicks and of the poults were removed. Three sites (anterior, mid, and posterior) of the small intestine were assigned a score from 0 to 4. Although discrete, intestinal lesions are not usually seen in either E. dispersa or E. mitis infections, other pathological signs are present. Visual intestinal scoring was based on epithelial color, fluid accumulation, and the overall general appearance of the intestine (serosal thickness, mucosal erosion, dilation, and similar factors).
An intestinal score of 0 was given to areas of the intestine that looked normal. Scores from 1 to 4 were used to rank the incidence and severity of the symptoms just described. The scores from each of the three intestinal sites were summed for analysis. The data were analyzed using analysis of variance procedures appropriate for a completely randomized design (Steel and Torrie, 1980). Since no interactions for treatment by trial were present, all data were pooled across trials. Due to heterogeneity of variance, the intestinal scores were transformed [log(+l)] prior to statistical analysis. Treatment differences were separated by the F-protected, least significant difference procedure. Pen was used as the experimental unit. RESULTS AND DISCUSSION
The results demonstrated that both E. mitis and E. dispersa infections can reduce growth as well as feed efficiency (Table 2). Although the species studied historically have been considered some of the least pathogenic, the present findings support those by others who reported that infections of E. mitis and E. dispersa can result in reduced growth and absorptive capacity (Joyner, 1958; Oluleye and Edgar, 1980; Ruff and Edgar, 1982; Allen, 1987). In the current study, Eimeria mitis and E. dispersa infections reduced gain 21 and 7% and reduced feed efficiency 14 and 4%. Eimeria mitis infection reduced (P<01) gain and the feed.gain ratio compared with uninfected birds, while E. dispersa infection only reduced (P<.05) gain. The lack of well-defined, discrete lesions in infections of E. mitis (Ruff and Edgar, 1982), E, dispersa (Madden and Ruff, 1979), and turkey coccidiosis in general (Jeffers and Bentley, 1980) made lesion scoring very difficult. Although gross intestinal lesions were not present in either E. mitis or E. dispersa infection, the authors did observe a reduced pigmentation, fluid accumulation, and a generalized abnormal appearance, resulting in relatively high intestinal scores (Table 2). Although the intestinal scores for birds infected with E. dispersa were similar to those for birds infected with E. mitis, the reduction in growth and feed efficiency due to E. dispersa infection was much less. No mortality occurred in any of the trials, although in some cases severe intestinal damage was noted.
Downloaded from http://ps.oxfordjournals.org/ at Rutgers University Libraries/Technical Services on June 1, 2015
Ingredient
2
1011
RESEARCH NOTE
TABLE 2. Effect of monensin on the growth and intestinal scores of chicks infected with Eimeria mitis and of poults infected with Eimeria dispersa Chicks infected with E. mitis
.O
Intestinal score
Gain
Feed:gain ratio
.00b 6.54" l-52 b
(£) 205 c 190d 205°
(g/g) 1.645 1.708 1.667
.76
3.79
.019
Intestinal score2 .51" 6.09" .44 b .51
"•"Means within columns with no common superscripts are significantly different (P<01). c,d Means within columns with no common superscripts are significantly different (P<.05). 1 NN=not infected, not medicated; IN=infected, not medicated; MON=infected, monensin-medicated. The chicks were fed diets containing 100 ppm of monensin; the poults were fed diets containing 60 ppm of monensin. ^Total score for the three intestinal sites, with a score of 12 being the highest possible. The intestinal scores were retransformed for table presentation. 3 Pooled.
Monensin was very effective in controlling the infections of both E. mitis and E. dispersa. The infected birds fed diets containing monensin had gains and feed:gain ratios similar (P>.10) to those of the uninfected birds. Monensin greatly reduced the incidence and severity of the intestinal abnormalities observed in the infected birds. No difference (P>.10) in intestinal scores was detected between the infected birds fed monensin and the uninfected, unmedicated birds. Oluleye and Edgar (1980) also reported that monensin was effective in controlling E. dispersa infections in turkeys. In summary, although E. mitis and E. dispersa are considered to be some of the lesspathogenic species of chicken and turkey coccidia, infection with either one can result in economic losses due to reductions in both growth and in feed efficiency. Monensin, when fed at 100 ppm to chicks or at 60 ppm to poults, provides excellent control of both E. mitis and E. dispersa. REFERENCES Allen, P. C , 1987. Physiological responses of chicken gut tissue to coccidial infection: Comparative effects of
Eimeria acervulina and Eimeria mitis on mucosal mass, carotenoid content, and brush border enzyme activity. Poultry Sci. 66:1306-1315. Jeffers, T. K., and E. J. Bentley, 1980. Monensin sensitivity of recent field isolates of turkey coccidia. Poultry Sci. 59:1722-1730. Joyner, L. P., 1958. Experimental fiwieria mitis infections in chickens. Parasitology 48:101-112. Madden, P. A., and M. D. Ruff, 1979. Eimeria dispersa, E. adenoeides, and E. meleagrimitis: intestinal mucosal disruption in turkeys as seen with scanning electron microscopy. J. Parasitol. 65:234-242. Novilla, M N., T. K. Jeffers, W. J. Griffing, and S. L. White, 1987. A redescription of the life cycle of Eimeria mitis, Tyzzer, 1929. J. Protozool. 34:87-92. Oluleye, O. R., and S. A. Edgar, 1980. Details of the life history and pathogenicity of the turkey coccidinm, Eimeria dispersa, Tyzzer 1929, and efficacy of four drugs for the control of the disease. Poultry Sci. 59(Suppl. 1):1569. (Abstr.) Ruff, M D., P. C. Augustine, and P. A. Madden, 1981. Eimeria meleagrimitis, E. adenoeides, and E. dispersa: severity of infection and changes in the intestinal mucosa of the turkey. Exp. Parasitol. 51:87-94. Ruff, M. D., and S. A. Edgar, 1982. Reduced intestinal absorption in broilers during Eimeria mitis infection. Am. J. Vet Res. 43:507-509. Steel, R.GX)., and J. H. Torrie, 1980. Principles and Procedures of Statistics: A Biometrical Approach. McGrawHill, Inc., New York, NY. Tyzzer, E. E., 1929. Coccidiosis in gallinaceous birds. Am. J. Hyg. 10:269-383.
Downloaded from http://ps.oxfordjournals.org/ at Rutgers University Libraries/Technical Services on June 1, 2015
c*i
(g) 288" 227 b 279" 9.38
at
NN IN MON SEM3
Feed:gain ratio X>
Gain
bo oo •£ 0\
Treatment1
Poults infected with E. dispersa