Medycyna Weterynaryjna - Summary Medycyna Wet. 63 (7), 805-809, 2007

Praca oryginalna Original paper

Coccidiosis is a protozoal disease that affects seve-ral different animal species including goats (1, 8, 14, 15, 20). The etiological agents are Eimeria and Iso-spora spp and their oocysts shed in the faeces of clini-cally affected or carrier animals. Ingestion of conta-minated feed and water and licking contaconta-minated hair coat play a crucial role in development of coccidiosis (22).

Acute clinic coccidiosis in goats commonly affects newly weaned goat kids kept in overcrowded con-dition (6, 13, 20, 21). Coccidiosis causes a significant economical loss due to retarded growth and death as a consequence of dysentery and anaemia (13, 20, 22). The mortality rate in goat kids could reach up to 40% (8). As cited by Jalila et al. (13), the mortality

rate can be even as high as 63%. The most common pathogenic coccidia species in goats are E. arloingi, E. ninakohlyokimovae, E. caprovina, E. christenseni, E. faurei and E. gilruthi (20, 22). In general, coocidia species are host-specific, and no cross-immunity occurs between species of coccidia (7, 8, 22, 27). Coc-cidiosis should be differentiated from the other causes of diarrhoea including Escherichia coli, Clostridium perfringens, Salmonella spp., and helminthiasis (22). Clinical sings, oocyst count in faeces, necropsy and pathological findings are used in differential diag-nosis (20, 22).

Anticoccidial drugs supported with oral or parente-ral fluids are recommended in treatment of acute clinical coccidiosis treatment (22). Chlortetracycline,

Toltrazuril treatment for acute clinical coccidiosis

in hair goat kids: clinical, pathological,

haematologic and biochemical findings

Department of Internal Medicine, *Department of Pathology, Faculty of Veterinary Medicine, Kirikkale University, Turkey

Öcal N., Yagci B. B., Duru S. Y., Kul O.

Toltrazuril treatment for acute clinical coccidiosis in hair goat kids: clinical, pathological, heamatologic and biochemical findings

Summary

This study was conducted to evaluate effectiveness and safety of oral administration of toltrazuril at a dose of 25 mg/kg/day for two consecutive days. The present study was carried out in a goat flock, reared in Kirikkale province of Turkey. Faeces samples were collected from 26 goat kids (6,716 ± 1.11 weeks old) exhibiting the clinical sings of acute clinic coccidiosis including dysentery, tenesmus, inappetence, and weakness. The diseased kids were randomly allocated into two groups: group I (n = 16); toltrazuril treatment and group II (n = 10); no treatment. An additional group was included as a healthy control (group III; n = 10). Prior to toltrazuril treatment, the number of oocysts was 8350 ± 4009 and 6295 ± 1490 in groups I and II, respectively. Eimeria arloingi (67.8%), E. ninakohlyakimovae (21.7%), E. apsheronica (6.4%), E. alijevi (4.1%) were determined in faeces. After toltrazuril treatment in group I, the number of oocysts reduced significantly down to 0 to 250 oocysts per gram faeces (p < 0.001). In contrast, the number of oocysts in untreated group (group II) did not declined, but even increased slightly. At the end of the study, all goat kids treated with toltrazuril (group I) recovered from clinical signs of coccidiosis. At necropsy carried out on two kids, one died prior to the beginning of the study and the other one died from group II, typical nodular coccidia lesions were encountered in intestinal tract. In goat kids with coccidiosis, a mild hypochromic macrositer anaemia developed which was determined on the basis of normal RBC count, increased MCV, lower Hb and PCV near the lower limit of the reference value. In addition, monocytosis and lymphopenia are also associated with coccidiosis in goat kids. Based upon clinical findings and a significant decrease in the number of oocysts in faeces (p < 0.001), it is concluded that oral administration toltrazuril at a dose of 25 mg/kg/day for two consecutive days provides an effective treatment for acute clinical coccidiosis in goat kids. As ALT, AST, BUN, and serum creatinine levels remain within the reference limits after the treatment; toltrazuril at the scheduled dose does not result in acute liver and kidney damage.

sent study was to evaluate the effectiveness of oral tol-trazuril treatment (25 mg/kg/day) for two consecutive days in hair goat kids suffering from acute clinical coccidiosis.

Material and methods

This study was carried out during a time period between March 15, 2005 and April 15, 2005 in a goat flock in Kirik-kale province of Turkey. The flock had 80 goat kids and 230 adult Turkish hair goats. Faeces were collected from 26 goat kids exhibiting clinical sings of acute clinic coccidiosis in-cluding dysentery, tenesmus, inappetence, and weakness. The mean age of the goat kids was 6,716 ± 1.11 weeks (ranging from 5 to 8.5 weeks) and the mean body weight was 9.482 ± 1.09 kg (ranging from 7 to 10 kg). The kids suffering from coccidiosis were randomly allocated into two groups: group I (n = 16); toltrazuril treatment (25 mg/kg/day for two con-secutive days) and group II (n = 10); control of treatment, no treatment. In addition, a third group (group III) (n = 10) was included as a healthy control. During the clinical phase of the study, goat kids in all three groups were kept separate. They were allowed to suckle and drink water.

The clinical, biochemical and haematological analyses as well as faeces examinations were conducted prior to and after toltrazuril treatment. Faecal samples were collected per rectum and the number of oocysts per gram of faeces was determined by the modified McMaster technique. Clinic coc-cidiosis was confirmed based on presence of 4000 or above oocysts per gram faeces. For sporulation, faeces samples were kept in 2.5% potassium dichromate at room temperature for two weeks. According to morphological characteristics, specie identification was made using 40 x the microscope objective (2, 8).

Blood samples were collected from the jugular vein for blood count and serum biochemistry. Total blood count and differential leucocytes were determined using a MS9 cell counter (MS9-3–Melet Schloesing Laboratoires, France). Serum ALT, AST (Valtek® Ltda. Nunoa Santiago-Chile), BUN, and serum creatinine (Teco

Diag-nostics, Anaheim-USA) were determined with spectrophotometric method (Shima-dzu UV-1208) following the instructions provided by the vendors.

Necropsy was carried out in two goat kids: one died just prior to the beginning of the study and another died in group II during the course of the study. Tissue

sam-and eyes. However, the body temperature, pulse sam-and respiratory rates were within normal limits. In toltra-zuril treatment (group I), the severity of diarrhoea as well as nasal and ocular discharges was decreased ap-proximately 24 hours after the first administration of toltrazuril. Moreover, the goat kids exhibited suckling behaviour more often. Twenty four hours after the second toltrazuril administration, clinical signs of coc-cidiosis disappeared in all kids except in 3 kids which still had mild diarrhoea. Faeces with an acceptable consistency and colour were noted. No clinical signs of drug toxicity or any unusual behaviour were obser-ved in toltrazuril treated group. In goat kids suffering from coccidiosis but received no treatment (group II), the severity of clinical signs worsened. Moreover, an incidence of death occurred.

Prior to treatment, faecal samples of the goat kids in groups I and II were positive for coccidia oocysts. There was no difference for the number of oocysts between groups I and II prior to toltrazuril treatment (p > 0.05). The number of oocysts in faecal samples was presen-ted in tab. 1. E. arloingi (67.8%), E. ninakohlyakimo-vae (21.7%), E apsheronica (6.4%), E alijevi (4.1%) were determined. No other parasite oocysts and larvae were encountered in faeces. After toltrazuril treatment in group I, the number of oocysts was significantly reduced (p < 0.001) down 0 to 250 per gram faeces. In contrast, the number of oocysts in group II did not change, but even increased slightly (tab. 1).

The pathological findings in the necropsied kids were similar. In gross examination, the jejunum and ileum had oedematous and wrinkled mucosa and hyperaemic submucosa. On the mucosal surface, grey to white nodular lesions of pin head size (1-2 mm in diameter) were observed. In some instances, several nodules conjoined to form larger nodules in ovoid shape (fig. 1). In histopathological examination, these

r e t e m a r a P _beforG_ero_tou_lp_rtaI_{zurli atfe}G_rr_to_olup_rtazI_{urli f(}G_risr_tou_cop_unII_{)t (sec}G_or_no_dup_coII_un)t r e b m u n t s y c o O 8350±4009,90a _90±92,96b _{6295±1490,80}a _{7070±1839,11}a

Tab. 1. The number of oocyst per gram faeces in goat kids with coccidiosis before and after treatment in group I and untreated group (group II) (–x ± SD)

Explanation: means with different superscripts within the same row differ significantly (p < 0.05)

nodular lesions appeared as of intestinal villi pro-truding towards the intestinal lumen. The epithelial surface of the protrusions had various developmental stages of Eimeria sp., such as schizont, microgameto-cyst, macrogametocyst and oocyst. The developmen-tal forms were mainly detected in the intestinal villus epithelia and intestinal glandular epithelia. Hyperaemic vessels, numerous macrophages, eosinophil leuco-cytes and lympholeuco-cytes infiltrations were features of the lamina propria. Hyperplasia, atrophy and necrosis were also noted in the intestinal glands (fig. 2).

The haematological parameters evaluated were pre-sented in tab. 2. In toltrazuril treatment group (group I), red blood cell (RBC) counts prior to toltrazuril treat-ment and after treattreat-ment were not significantly dif-ferent (p < 0.05). However, toltrazuril treatment did not lower the RBC count down to a level comparable to that of control as RBC counts in group III (health control) was significantly lower than both counts of group I, prior to and after treatment (p < 0.05). After toltrazuril treatment, the leukocyte count declined significantly (p < 0.05) down to a level comparable to that of health control kids (tab. 2).

The mean corpuscular volume (MCV) was not sig-nificantly different between both measurements of group I, prior to and after treatment. However, both measurements of group I were significantly higher than that of group III (p < 0.001).

Packed cell volume (PCV) value was not signifi-cantly different between both measurements of group I, prior to and after treatment. Both measurements in group I were not significantly different compared to group III, either.

The mean haemoglobin (Hb) value in group I was significantly higher after toltrazuril treatment compa-red to that measucompa-red before treatment (p < 0.05). Both measurements of group I were significantly lower than that of group III (p < 0.01).

The mean percent lymphocyte (LYMP) was not sig-nificantly different between both measurements of group I, prior to and after treatment. For LYMP, group III was significantly different both counts of group I, prior to toltrazuril treatment (p < 0.05) and after treat-ment (p < 0.01).

The mean percent monocyte (MONO) was not sig-nificantly different between both measurements of group I, prior to and after treatment. On the other hand, the percent MONO in group III was significantly lower than both counts of group I (p < 0.01).

Tab. 3: Biochemical parameters in goat with coccidiosis before and after toltrazuril treatment in group I and in healthy controls (group III) (–x ± SD)

r e t e m a r a P Gro_tou_lp_rtaI_zb_ue_rf_liore G_tro_olup_rtaI_zua_rtf_lier GroupIII )l / U ( T L A 17,23±2,86a _17,82±2,86a _25,8±8,20b )l / U ( T S A 210,07±11,57a _{219,52±13,54}b _{224,25±12,00}b )l /l o m m ( N U B 11,18±1,98 11,41±2,26 10,93±2,17 e n i n it a e r C )l /l o m µ ( 86,63±7,96 85,75±6,19 78,68±7,51 Explanation: as in tab. 1. Tab. 2. Hematological parameters in goat with coccidiosis

before and after toltrazuril treatment in group I and in healthy controls (group III) (–x ± SD)

r e t e m a r a P Gro_tou_lp_rtaI_zb_ue_rf_liore G_tro_olup_rtaI_zua_rtf_lier GroupIII ( C B R × 01 12_{/)l 17,05±2,02}a _16,71±2,76a _15,69±0,82b ( C B W × 01 9_{/)l 17,38±5,39}a _13,98±3,02b _12,23±1,94b )l f( V C M 22,77±1,41a _22,73±1,30a _18,43±0,88b )l /l ( V C P 0,224±0,027 0,228±0,025 0,227±0,018 )l / g ( g H 66,6±7,6a _67,6±7,8a _81,2±12,0b ) % ( P M Y L 41,49±9,34a _39,96±9,05a _48,06±4,43b ) % ( O N O M 19,46±3,20a _20,76±4,23a _6,53±1,54b ) % ( T U E N 39,02±9,54 39,30±8,52 38,65±4,57 Explanation: as in tab. 1.

Fig. 2. Oocysts (thick arrow) and leukocyte and macrophage infiltrations (small arrows) in the lamina propria of the jejunum, (H+E), ×180

Fig. 1. Numerous grey to white foci (arrows) on the mucosa of the jejunum

cantly different among groups and unchanged after treatment in group I (p > 0.05).

Acute clinical coccidiosis usually affects weaning kids kept under inappropriate care and condition (21). The disease is often associated with anaemia, dysen-tery, tenesmus, retarded growth, and death (22). The goat kids included in this study had been kept in an inappropriate and crowded condition. During the clinical phase of the study, the geographical region, where the goat kids reared, received lots of rain, which was possibly an additional stress factor (21). Goat kids with acute clinical coccidiosis in the present study were clinically characterised by severe diarrhoea, weakness, weight loss and nasal and lacrimal discharges in some kids. Jalila et al. (13) proposed that the incidence of clinical coccidiosis is often higher in kids of 6-16 weeks of age, during which passive immunity gradually weakened. Such a proposition is also supported by the present study as the diseased goat kids were 5-8 weeks of age. Jalila et al. (13) also indicated that adult goats should be considered as the source of infection. Acute clinical coccidiosis was diagnosed based upon clinical signs, presence of a large number of oocysts in faeces, and pathological findings (20, 22). In patho-logical examination, no other lesions or parasite were observed other than typical pathological coccidia lesions in the intestine.

Balicka-Ramisz (5) reported that administration of toltrazuril at a dose of 20 mg/kg twice with a week interval, is successful in the treatment of coccidiosis. In another study, Balicka-Ramisz (4) found oral admi-nistration of toltrazuril at a dose of 20 mg/kg body weight was effective in the treatment of coccidiosis in goats. In a previous study, Frandsen et al. (11) repor-ted a single dose of 20 mg/kg toltrazuril was not effec-tive in the treatment of goat coccidiosis. In the present study, we found that oral administration of toltrazuril at a dose of 25 mg/kg/day for two consecutive days is successful in the treatment of acute clinical coccidio-sis in goat kids as the number of oocysts in faeces was remarkably reduced and the clinical conditions of the kids improved after the toltrazuril treatment. In addi-tion, no clinical signs of drug toxicity or any unusual behaviour were observed. In contrast, the severity of clinical signs in kids of the untreated group increased.

in RBC count most likely occurred due to an erytroid--regenerative response as a consequence of dysentery (16). In the present study, expected anaemia was not as prominent that might be related to clinical stage of coccidiosis. Besides, it is known that anaemia is not always determined based only on the RBC count (29). In this study, the leukocyte count after toltrazuril treatment significantly declined and became com-parable to that of health control. Thus, the toltrazuril effect is also reflected as a reduction in leukocytosis, which is an ongoing feature of coccidiosis (9, 26).

In toltrazuril treatment group, MCV was not diffe-rent between the values measured prior to and after treatment. However, both measurements were signifi-cantly higher than that of group III (p < 0.01). This condition was interpreted as macrocytosis related to an eryhtroid-regenerative response to intestinal blood loss (16).

There was no significant difference between values measured prior to and after toltrazuril treatment for PCV. Compared to both values of group I, PCV in healthy controls was not significantly different, either. However, PCV was slightly lower prior to toltrazuril treatment and close to the lower limit of the reference value (17). This condition is considered as an indica-tion of mild anaemia or a pre-disposiindica-tion to anaemia. According to Tvedten and Weiss (29), the PCV value of 20-26% may indicate presence of mild anaemia.

The mean Hb values measured both prior to and after toltrazuril treatment were significantly lower (hypo-chromia) compared to that of healthy controls (81.2 g/l) (p < 0.01). This condition might be related to intesti-nal bleeding in coccidiosis or to feeding practice of the goat kids, which is suckling only.

The percent LYMP in diseased kids (41.49%) was not significantly changed following toltrazuril treat-ment (39.96%); however, both values were lower (lymphopenia) compared to healthy controls (48.06%) and compared to the reference value (50-70%). Lym-phopenia can be explained by massive migration of lymphocytes to inflammation sites in response to in-flammatory mediators (25).

The percent MONO in diseased kids was not sig-nificantly changed following toltrazuril treatment; however, both values were higher (monocytosis)

com-pared to that of healthy controls (6.53%) and the reference value (0-4%). This finding may support Stockham (25) who suggested that monocytosis could occur in the early phase of inflammatory response.

The percent NEUT in diseased kids was not changed following the toltrazuril treatment; however, the percent neutrophil in healthy controls was lower although the difference was not significant. The per-cent neutrophil is expected to be shifted to the left in acute infections; however, this was not prominent in the present study, and the percent neutrophil was within the limits of the reference value (30-48%). This condition is thought to be related to accumulation of neutrophils to inflammation sites during the early phase of inflammation (25) (fig. 2).

Turk and Casteel (28) reported that the serum levels of ALT and AST are increased in acute hepatic dege-nerations. In the present study, ALT was not signifi-cantly different before and after toltrazuril treatment and within the limits of the reference value. The AST level in diseased kids (210.07 U/l) slightly increased after toltrazuril treatment, which, however, was not significantly different from that of healthy controls (224.25 U/l). Thus, we thought that toltrazuril does not cause an acute damage in the liver, which is the main route of toltrazuril excretion (3).

The blood BUN and serum creatinine levels can elevate in response to increase in protein catabolism caused by hepatotoxins, myotoxins, and the other necrotisan agents as well as nephrotoxic drugs (28). In the present study, no difference was found between diseased and control kids as well as between before and after toltrazuril treatment. This finding may indi-cate that toltrazuril (25 mg/kg/day for two consecu-tive days) does not cause acute kidney damage, which is another important route of excretion.

Conclusion

Based upon clinical findings and a significant de-crease in oocyst count in faeces, we concluded that oral dose of toltrazuril given 25 mg/kg/day for two consecutive days provides an effective treatment in acute clinical coccidiosis in goat kids. As evidenced by ALT, AST, BUN, and serum creatinine levels com-parable to reference values after toltrazuril treatment at the above mentioned dose, we also suggest that tol-trazuril does not cause acute hepatic and nephrotic damage.

Mild hypochromic macrositer anaemia develops in goat kids with coccidiosis evidenced by normal RBC count, increased MCV, lower Hb and PCV which is close to the lower reference limit. In addition, mono-cytosis and lymphopenia are also associated with coc-cidiosis in goat kids.

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