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Prevalence, hematology, and treatment of balantidiasis among donkeys in and around Lahore, Pakistan
Veterinary Parasitology 196 (2013) 203–205
Contents lists available at SciVerse ScienceDirect
Veterinary Parasitology journal homepage: www.elsevier.com/locate/vetpar
Short communication
Prevalence, hematology, and treatment of balantidiasis among donkeys in and around Lahore, Pakistan A. Khan, M.S. Khan, M. Avais, M. Ijaz, M.M. Ali ∗ , T. Abbas Department of Clinical Medicine and Surgery, Faculty of Veterinary Science, University of Veterinary and Animal Sciences, Lahore 54000, Punjab, Pakistan
a r t i c l e
i n f o
Article history: Received 23 April 2012 Received in revised form 17 January 2013 Accepted 18 January 2013 Keywords: Balantidium coli Donkey Secnidazole Prevalence Nigella sativa
a b s t r a c t The prevalence of Balantidium coli among donkeys in Lahore and adjoining areas was surveyed and a trial conducted to determine the efficacy of two antiprotozoal drugs: secnidazole (Dysen Forte) and Kalonji (Nigella sativa). Four-hundred donkeys were examined, and 73 (18.3%) were found positive for Balantidium coli. A slight decrease in PCV and an increase in Hb values of infected donkeys were found after antiprotozoal treatment. Secnidazole was 89.5% effective for the treatment of equine balantidiasis compared to 40.0% for Nigella sativa. This is the first report of balantidiasis in equines from Pakistan. It is not known if balantidiasis is an emerging problem in equines or whether it is a newly reported infection. © 2013 Elsevier B.V. All rights reserved.
1. Introduction Balantidium coli has a wide variety of hostsincluding rodents, pigs (wild and domestic), camels, and primates, both human and nonhuman (Taylor et al., 2007). Considerable information is available on B. coli in cattle (Bilal et al., 2009). It is less commonly found in dogs (Barr, 2005; Taylor et al., 2007) and equines (Bowman, 2009; Headley et al., 2008).B. coli has been considered a non-significant pathogen,especially in equines, and investigations of B. coli have not been actively conducted in even intensive equinerearing areas. B. coli is ofzoonotic importance,especially with respect to infections in pigs (Schuster et al., 2008). It has also been characterized as an emerging protozoan pathogen (Garcia, 2008; Schuster et al., 2008). In view of the importance of donkeys in developing countries, the potential for a
∗ Corresponding author at: Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore, Pakistan. Tel.: +92 3336252189. E-mail address: [email protected] (M.M. Ali). 0304-4017/$ – see front matter © 2013 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.vetpar.2013.01.017
zoonotic relationship between donkeys and humans exists, although this has not been documented. The primary aim of this study was to survey donkeys for B. coli infection to determine its significance in this group of equines. 2. Materials and methods Approximately 5 g of feces was taken directly from the rectum of 400 donkeys of mixed age and sex (M = 210; F = 190). The samples were placed in self-sealing polythene bags and held at 4 ◦ C for analysis within 24 h. A direct smear and sedimentation were used to confirm the presence of B. coli cysts,which were counted with a McMaster slide. 2.1. Chemotherapeutic trial Of 73 donkeys positive for B. coli, 30 were randomly selected and separated into three groups of 10 (A–C). A fourth group (D) comprised 10 uninfected donkeys. Group A was treated with the nitroimidazole, secnidazole (Dysen Forte, NabiQasim, Pvt. Ltd., Pakistan) at a single oral dose of 10 mg/kg and Group B with Nigella sativa (Kalonji) at a daily oral dose of 200 mg/kg for five days. Group C (untreated)
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A. Khan et al. / Veterinary Parasitology 196 (2013) 203–205
Table 1 Age and sex based prevalence of B. coli in donkeys. Age groups
≤2 yeara 2–5 yeara Above 5 yeara a
Malea
Femalea
No. examined
No. positive
prevalence (%)
No. examined
No. positive
Prevalence (%)
83 97 30
18 14 4
21.69 14.43 13.33
63 79 48
15 14 8
23.81 17.72 16.67
Indicates non-significant difference (P > 0.05) among different age and sex groups.
Table 2 Balantidium coli cyst counts in treated and control groups of donkeys (mean ± SD). Group
Treatment
Aa Ba Ca Da
Secnidazole Nigella sativa Positive Control Negative Control
Mean (n = 10) cyst count per g Day 0a
a
242.2 ± 83.72 213.9 ± 87.1 210.7 ± 90.79 0.0
Day 3a 146 ± 51.42 188.2 ± 82.66 213.1 ± 90.58 0.0
Day 7a
Day 14a
57.6 ± 20.16 162.5 ± 76.17 222.3 ± 90.48 0.0
25.4 ± 8.33 128.3 ± 70.48 233.1 ± 89.61 0.0
Significant difference among groups at various days (P < 0.05).
was designated as positive control, and the animals in Group D as negative controls. Fecal samples were collected on day 0 (pre-treatment) and on days 3, 7, and 14 following initiation of treatment for all groups. The efficacy of the drugs was assessed on the basis of reduction in cyst count and was calculated according to the formula of Varady et al. (2004): Pretreatment CPG − Post treatment CPG × 100 Pretreatment CPG In hematological studies, hemoglobin estimation (g/dl) by Sahli’s method (Coles, 1986) and packed cell volume (PCV) by the method of Maxwell and Wintrobe (1929) were carried out. Blood samples (5 ml) were collected from all groups with disposable 18G sterilized hypodermic needles in an EDTA coated vacutaineron day 0 (pre-treatment) and on days 3, 7, and 14 post-treatment. Prevalence of B. coli infection was analyzed by 2 test using the statistical software package STATA 9.1 (College Station, TX 77845, USA) whereas data on hematology was analyzed through one way ANOVA. 3. Results and discussion Data on age and sex-based prevalence is shown in Table 1. The prevalence of B. coli was not significantly different (P > 0.05) among age groups and sexes. Of the 400 donkeys, 73 (18.3%) tested positive for B. coli, which corresponds closely to the findings of Bilal (2006) and Tarrar et al. (2008), who reported prevalence of B. coli in cattle and buffalo around the banks of the River Ravi, Lahore to be 25% and 20%, respectively. The cyst counts in Groups A and B were significantly lower compared togroup C (P < 0.05, Table 2). Secnidazole showed higher efficacy compared to Nigella sativa, and neither treatment was associated with post-treatment side effects (Table 3). These findings confirm efficacy of secnidazolereported by Bilal (2009) and Tarrar et al. (2008) of 75% and 83%,respectively. In current study, Nigella sativa
showed poor efficacy, contrary to Baghdadi and Al-Mathal (2011), who reported good anti-parasitic efficacy when used at the dose of 200–400 mg/kg body weight. Increase in hemoglobin and decrease in PCV values among groups were observed at 3, 7, and 14 days posttreatment (P < 0.05, Table 4). The slightly lower value of hemoglobinand the higher PCV inGroups A, B, and C on day 0 and the initial days post-treatment may be due to pathological lesions, bloody diarrhea, severe hemorrhage, or digestive disturbances. No reports were found regarding the hematological values in B. coli infected equines, but Miller (2003) reported mild anemia, marked leukocytosis, and mild hypo-albuminemia in B. coli positive chimpanzees.B. coli infection occurs in most developing countries where water sources are contaminated by sewage (Schuster et al., 2008). This study has shown that there is substantial infection in donkeys, presumably arising from contaminated water. Strategies should be made to eliminate the transmission factors in equines, such as provision of clean drinking water and housing equines separately from other domestic animals to avoid transmission. As equine infections have not been reported previously, it is impossible to determine if this is an emerging disease or one that is newly reported. Secnidazole is the drug of choice for the treatment of balantidiasis in donkeys.
Table 3 Efficacy of secnidazole and Nigellasativa in reducing B. coli cysts in donkeys at three time intervals post-infection based on the formula given by Varady et al. (2004). Groups
Treatment
Aa Ba
Secnidazole Nigella sativa
a
Significant difference (P < 0.05).
% efficacy days post-infection 3
7
14
39.72 12.02
76.22 24.03
89.51 40.02
A. Khan et al. / Veterinary Parasitology 196 (2013) 203–205
205
Table 4 Hemoglobinand PCV intreatment groups of donkeys infected with Balantidium coli at four time intervals. Groups
Treatment
Day0a
Day3a
Mean Hb (g/dl) ± S.D. Aa Ba Ca Da a
Secnidazole Nigellasativa Positive control Negative control
10.61 10.55 10.61 11.01
± ± ± ±
0.07 0.13 0.15 0.06
Mean PCV (%) ± S.D. 31.37 31.48 31.47 30.14
± ± ± ±
0.27 0.22 0.24 0.24
Day7a
Mean Hb (g/dl) ± S.D. 10.61 10.54 10.46 11.39
± ± ± ±
0.076 0.13 0.15 0.10
Mean PCV (%) ± S.D. 31.47 31.48 31.67 30.19
± ± ± ±
0.27 0.22 0.24 0.25
Day14a
Mean Hb (g/dl) ± S.D. 10.68 10.64 10.01 11.03
± ± ± ±
0.097 0.127 0.15 0.02
Mean PCV (%) ± S.D. 31.20 31.61 31.88 30.77
± ± ± ±
0.24 0.14 0.24 0.29
Mean Hb (g/dl) ± S.D. 10.79 10.69 10.00 10.91
± ± ± ±
0.094 0.126 0.15 0.09
Mean PCV (%) ± S.D. 31.12 31.44 31.87 30.50
± ± ± ±
0.16 0.13 0.24 0.31
Significant difference among groups at various days (P < 0.05).
References Baghdadi, H.B., Al-Mathal, E.M., 2011. Anti-coccidial activity of Nigella sativa L. J. Food Agric. Environ. 9, 10–17. Barr, S.C., 2005. Balantidiasis. In: Green, C.E. (Ed.), Infectious Diseases of the Dog and Cat., 3rd ed. Elsevier, Saunders, pp. 744–745. Bilal, C.Q., Khan, M.S., Avais, M., Ijaz, M., Khan, J.A., 2009. Prevalence and chemotherapy of Balantidium coli in cattle in the River Ravi region Lahore (Pakistan). Vet. Parasitol. 163, 15–17. Bowman, D.D., 2009. Georges’ parasitology for veterinarian, 9th ed. W.B Saunders Co, Philadelphia. Coles, E.H., 1986. Veterinary Clinical Pathology, 4th ed. W.B. Saunders Co, Philadelphia. Garcia, L.S., 2008. Balantidium coli. In: Khan, N.A. (Ed.), Emerging Protozoan Pathogens. Taylor and Francis, New York, pp. 353–366.
Headley, S.A., Kummala, E., Sukura, A., 2008. Balantidium coli-infection in a Finnish horse. Vet. Parasitol. 158, 129–132. Maxwell, M., Wintrobe, M.D., 1929. The volume and hemoglobin content of the red blood corpuscles. Amer. J. Med. Sci. 177, 513–523. Miller, G., 2003. Blood changes in Balantidium coli infected chimpanzee. Department of Veterinary Pathology, The Armed Force Institute of Pathology, Bethesda, U.S.A, Case-IV-N pp. 21–98. Schuster, F.L., Ramirez-Avila, Lynn., 2008. Current world status of Balantidium coli. Clin. Microbiol. Rev. 21, 626–638. Tarrar, M.A., Khan, M.S., Pervez, K., Ashraf, K., Khan, J.A., Rehman, Z.U., 2008. Detection and chemotherapy of Balantidium coli in buffaloes around Lahore Pakistan. Pak. J. Agric. Sci. 45, 163–166. Taylor, M.A., Coop, R.L., Wall, R.L., 2007. Veterinary Parasitology, 3rd ed. Blackwell, Ames, 874. Varady, M., Konigova, A., Corba, J., 2004. A field study to evaluate the efficacy of fenbendazole on 9 stud farms. Vet. Med. Czech. 49, 42–46.
Contents lists available at SciVerse ScienceDirect
Veterinary Parasitology journal homepage: www.elsevier.com/locate/vetpar
Short communication
Prevalence, hematology, and treatment of balantidiasis among donkeys in and around Lahore, Pakistan A. Khan, M.S. Khan, M. Avais, M. Ijaz, M.M. Ali ∗ , T. Abbas Department of Clinical Medicine and Surgery, Faculty of Veterinary Science, University of Veterinary and Animal Sciences, Lahore 54000, Punjab, Pakistan
a r t i c l e
i n f o
Article history: Received 23 April 2012 Received in revised form 17 January 2013 Accepted 18 January 2013 Keywords: Balantidium coli Donkey Secnidazole Prevalence Nigella sativa
a b s t r a c t The prevalence of Balantidium coli among donkeys in Lahore and adjoining areas was surveyed and a trial conducted to determine the efficacy of two antiprotozoal drugs: secnidazole (Dysen Forte) and Kalonji (Nigella sativa). Four-hundred donkeys were examined, and 73 (18.3%) were found positive for Balantidium coli. A slight decrease in PCV and an increase in Hb values of infected donkeys were found after antiprotozoal treatment. Secnidazole was 89.5% effective for the treatment of equine balantidiasis compared to 40.0% for Nigella sativa. This is the first report of balantidiasis in equines from Pakistan. It is not known if balantidiasis is an emerging problem in equines or whether it is a newly reported infection. © 2013 Elsevier B.V. All rights reserved.
1. Introduction Balantidium coli has a wide variety of hostsincluding rodents, pigs (wild and domestic), camels, and primates, both human and nonhuman (Taylor et al., 2007). Considerable information is available on B. coli in cattle (Bilal et al., 2009). It is less commonly found in dogs (Barr, 2005; Taylor et al., 2007) and equines (Bowman, 2009; Headley et al., 2008).B. coli has been considered a non-significant pathogen,especially in equines, and investigations of B. coli have not been actively conducted in even intensive equinerearing areas. B. coli is ofzoonotic importance,especially with respect to infections in pigs (Schuster et al., 2008). It has also been characterized as an emerging protozoan pathogen (Garcia, 2008; Schuster et al., 2008). In view of the importance of donkeys in developing countries, the potential for a
∗ Corresponding author at: Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore, Pakistan. Tel.: +92 3336252189. E-mail address: [email protected] (M.M. Ali). 0304-4017/$ – see front matter © 2013 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.vetpar.2013.01.017
zoonotic relationship between donkeys and humans exists, although this has not been documented. The primary aim of this study was to survey donkeys for B. coli infection to determine its significance in this group of equines. 2. Materials and methods Approximately 5 g of feces was taken directly from the rectum of 400 donkeys of mixed age and sex (M = 210; F = 190). The samples were placed in self-sealing polythene bags and held at 4 ◦ C for analysis within 24 h. A direct smear and sedimentation were used to confirm the presence of B. coli cysts,which were counted with a McMaster slide. 2.1. Chemotherapeutic trial Of 73 donkeys positive for B. coli, 30 were randomly selected and separated into three groups of 10 (A–C). A fourth group (D) comprised 10 uninfected donkeys. Group A was treated with the nitroimidazole, secnidazole (Dysen Forte, NabiQasim, Pvt. Ltd., Pakistan) at a single oral dose of 10 mg/kg and Group B with Nigella sativa (Kalonji) at a daily oral dose of 200 mg/kg for five days. Group C (untreated)
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A. Khan et al. / Veterinary Parasitology 196 (2013) 203–205
Table 1 Age and sex based prevalence of B. coli in donkeys. Age groups
≤2 yeara 2–5 yeara Above 5 yeara a
Malea
Femalea
No. examined
No. positive
prevalence (%)
No. examined
No. positive
Prevalence (%)
83 97 30
18 14 4
21.69 14.43 13.33
63 79 48
15 14 8
23.81 17.72 16.67
Indicates non-significant difference (P > 0.05) among different age and sex groups.
Table 2 Balantidium coli cyst counts in treated and control groups of donkeys (mean ± SD). Group
Treatment
Aa Ba Ca Da
Secnidazole Nigella sativa Positive Control Negative Control
Mean (n = 10) cyst count per g Day 0a
a
242.2 ± 83.72 213.9 ± 87.1 210.7 ± 90.79 0.0
Day 3a 146 ± 51.42 188.2 ± 82.66 213.1 ± 90.58 0.0
Day 7a
Day 14a
57.6 ± 20.16 162.5 ± 76.17 222.3 ± 90.48 0.0
25.4 ± 8.33 128.3 ± 70.48 233.1 ± 89.61 0.0
Significant difference among groups at various days (P < 0.05).
was designated as positive control, and the animals in Group D as negative controls. Fecal samples were collected on day 0 (pre-treatment) and on days 3, 7, and 14 following initiation of treatment for all groups. The efficacy of the drugs was assessed on the basis of reduction in cyst count and was calculated according to the formula of Varady et al. (2004): Pretreatment CPG − Post treatment CPG × 100 Pretreatment CPG In hematological studies, hemoglobin estimation (g/dl) by Sahli’s method (Coles, 1986) and packed cell volume (PCV) by the method of Maxwell and Wintrobe (1929) were carried out. Blood samples (5 ml) were collected from all groups with disposable 18G sterilized hypodermic needles in an EDTA coated vacutaineron day 0 (pre-treatment) and on days 3, 7, and 14 post-treatment. Prevalence of B. coli infection was analyzed by 2 test using the statistical software package STATA 9.1 (College Station, TX 77845, USA) whereas data on hematology was analyzed through one way ANOVA. 3. Results and discussion Data on age and sex-based prevalence is shown in Table 1. The prevalence of B. coli was not significantly different (P > 0.05) among age groups and sexes. Of the 400 donkeys, 73 (18.3%) tested positive for B. coli, which corresponds closely to the findings of Bilal (2006) and Tarrar et al. (2008), who reported prevalence of B. coli in cattle and buffalo around the banks of the River Ravi, Lahore to be 25% and 20%, respectively. The cyst counts in Groups A and B were significantly lower compared togroup C (P < 0.05, Table 2). Secnidazole showed higher efficacy compared to Nigella sativa, and neither treatment was associated with post-treatment side effects (Table 3). These findings confirm efficacy of secnidazolereported by Bilal (2009) and Tarrar et al. (2008) of 75% and 83%,respectively. In current study, Nigella sativa
showed poor efficacy, contrary to Baghdadi and Al-Mathal (2011), who reported good anti-parasitic efficacy when used at the dose of 200–400 mg/kg body weight. Increase in hemoglobin and decrease in PCV values among groups were observed at 3, 7, and 14 days posttreatment (P < 0.05, Table 4). The slightly lower value of hemoglobinand the higher PCV inGroups A, B, and C on day 0 and the initial days post-treatment may be due to pathological lesions, bloody diarrhea, severe hemorrhage, or digestive disturbances. No reports were found regarding the hematological values in B. coli infected equines, but Miller (2003) reported mild anemia, marked leukocytosis, and mild hypo-albuminemia in B. coli positive chimpanzees.B. coli infection occurs in most developing countries where water sources are contaminated by sewage (Schuster et al., 2008). This study has shown that there is substantial infection in donkeys, presumably arising from contaminated water. Strategies should be made to eliminate the transmission factors in equines, such as provision of clean drinking water and housing equines separately from other domestic animals to avoid transmission. As equine infections have not been reported previously, it is impossible to determine if this is an emerging disease or one that is newly reported. Secnidazole is the drug of choice for the treatment of balantidiasis in donkeys.
Table 3 Efficacy of secnidazole and Nigellasativa in reducing B. coli cysts in donkeys at three time intervals post-infection based on the formula given by Varady et al. (2004). Groups
Treatment
Aa Ba
Secnidazole Nigella sativa
a
Significant difference (P < 0.05).
% efficacy days post-infection 3
7
14
39.72 12.02
76.22 24.03
89.51 40.02
A. Khan et al. / Veterinary Parasitology 196 (2013) 203–205
205
Table 4 Hemoglobinand PCV intreatment groups of donkeys infected with Balantidium coli at four time intervals. Groups
Treatment
Day0a
Day3a
Mean Hb (g/dl) ± S.D. Aa Ba Ca Da a
Secnidazole Nigellasativa Positive control Negative control
10.61 10.55 10.61 11.01
± ± ± ±
0.07 0.13 0.15 0.06
Mean PCV (%) ± S.D. 31.37 31.48 31.47 30.14
± ± ± ±
0.27 0.22 0.24 0.24
Day7a
Mean Hb (g/dl) ± S.D. 10.61 10.54 10.46 11.39
± ± ± ±
0.076 0.13 0.15 0.10
Mean PCV (%) ± S.D. 31.47 31.48 31.67 30.19
± ± ± ±
0.27 0.22 0.24 0.25
Day14a
Mean Hb (g/dl) ± S.D. 10.68 10.64 10.01 11.03
± ± ± ±
0.097 0.127 0.15 0.02
Mean PCV (%) ± S.D. 31.20 31.61 31.88 30.77
± ± ± ±
0.24 0.14 0.24 0.29
Mean Hb (g/dl) ± S.D. 10.79 10.69 10.00 10.91
± ± ± ±
0.094 0.126 0.15 0.09
Mean PCV (%) ± S.D. 31.12 31.44 31.87 30.50
± ± ± ±
0.16 0.13 0.24 0.31
Significant difference among groups at various days (P < 0.05).
References Baghdadi, H.B., Al-Mathal, E.M., 2011. Anti-coccidial activity of Nigella sativa L. J. Food Agric. Environ. 9, 10–17. Barr, S.C., 2005. Balantidiasis. In: Green, C.E. (Ed.), Infectious Diseases of the Dog and Cat., 3rd ed. Elsevier, Saunders, pp. 744–745. Bilal, C.Q., Khan, M.S., Avais, M., Ijaz, M., Khan, J.A., 2009. Prevalence and chemotherapy of Balantidium coli in cattle in the River Ravi region Lahore (Pakistan). Vet. Parasitol. 163, 15–17. Bowman, D.D., 2009. Georges’ parasitology for veterinarian, 9th ed. W.B Saunders Co, Philadelphia. Coles, E.H., 1986. Veterinary Clinical Pathology, 4th ed. W.B. Saunders Co, Philadelphia. Garcia, L.S., 2008. Balantidium coli. In: Khan, N.A. (Ed.), Emerging Protozoan Pathogens. Taylor and Francis, New York, pp. 353–366.
Headley, S.A., Kummala, E., Sukura, A., 2008. Balantidium coli-infection in a Finnish horse. Vet. Parasitol. 158, 129–132. Maxwell, M., Wintrobe, M.D., 1929. The volume and hemoglobin content of the red blood corpuscles. Amer. J. Med. Sci. 177, 513–523. Miller, G., 2003. Blood changes in Balantidium coli infected chimpanzee. Department of Veterinary Pathology, The Armed Force Institute of Pathology, Bethesda, U.S.A, Case-IV-N pp. 21–98. Schuster, F.L., Ramirez-Avila, Lynn., 2008. Current world status of Balantidium coli. Clin. Microbiol. Rev. 21, 626–638. Tarrar, M.A., Khan, M.S., Pervez, K., Ashraf, K., Khan, J.A., Rehman, Z.U., 2008. Detection and chemotherapy of Balantidium coli in buffaloes around Lahore Pakistan. Pak. J. Agric. Sci. 45, 163–166. Taylor, M.A., Coop, R.L., Wall, R.L., 2007. Veterinary Parasitology, 3rd ed. Blackwell, Ames, 874. Varady, M., Konigova, A., Corba, J., 2004. A field study to evaluate the efficacy of fenbendazole on 9 stud farms. Vet. Med. Czech. 49, 42–46.