Chemoprophylaxis of coccidiosis in lambs with a single oral dose of toltrazuril

The prophylactic efficacy of a single oral dose of toltrazuril against coccidiosis (mixed Eimeria infections) in naturally infected lambs was evaluated in two experiments. Toltrazuril at 20 mg kg-1, given on Day 7 or Day 10 after turnout on pasture, proved to be highly efficacious in preventing clinical coccidiosis under Norwegian conditions. Toltrazuril reduced the oocyst output to very low levels, prevented the development of diarrhoea and improved weight gain during the first 4-5 weeks after treatment. Treatment on Day 7 was superior to treatment on Day 10 with respect to improving weight gain and preventing the development of soft faeces. Lambs treated with toltrazuril on Day 7 seemed to be as immune as untreated lambs to natural reinfections with coccidia later in the grazing season. In one of the experiments, natural infections with the nematode Nematodirus battus produced a coccidiosis-like disease in some lambs simultaneously with the outbreak of coccidiosis.     

Life cycle, host restriction and longevity of Cyrilia nili (Haemogregarina nili Wenyon, 1909) n. comb.

Morphometric and morphological observations on C. nili in an experimentally infected fish (using the leech vector Batracobdeloides tricarinata for the first time) revealed two successive types of merogonic cycles occurring within the fish erythrocytes. In the first cycle large meronts produced eight small merozoites each while in the second cycle smaller meronts produced four merozoites each. Merozoites of the second cycle were destined to become gamonts. The gamonts were somewhat larger than merogonic stages and comprised the majority of the parasitic stages during parasitemia for up to 8 months. In the leech crop, gamonts released from the fish erythrocytes, became associated in syzygy and fused. The formed zygote underwent sporogony within the intestinal tissues and up to 60 sporozoites were produced from an ovoid or round oocyst. The produced sporozoites migrated toward the salivary and the proboscis tissues. The parasite survived in the leech over a period of 8 months involving 5 meals after the initial meal provided the fasting period did not exceed 75-105 days. Survival was attributed to residual stages in the proboscis and salivary tissues. Cross transmission experiments between fishes via B. tricarinata revealed that the fish haemogregarines were not host specific. Previously described African fresh water fish haemogregarines were indistinguishable and on the basis of the current results, they are regarded as a single species Cyrilia nili.     

Efficacy of toltrazuril in the prevention of coccidiosis in naturally infected lambs on pasture

The efficacy of toltrazuril (Bay Vi 9142) in preventing ovine coccidiosis due to an infection acquired immediately after turnout on pasture was evaluated by comparing the faecal consistency, weight gain, and oocyst output of treated and untreated lambs in 3 trials. The lambs were either given a single treatment with toltrazuril at 15 or 20 mg/kg, or they were given a dose of 10 mg/kg on 2 separate days. A single treatment with toltrazuril at 20 mg/kg on day 10 after turnout on pasture almost completely prevented coccidiosis in 2 trials. In a third trial the acute phase of a severe Nematodirus battus infection coincided with the outbreak of coccidiosis, and thus partly masked the clinical effect of the anticoccidial treatment. In lambs treated with toltrazuril at 15 mg/kg on day 10 after turnout, the coccidial infection caused a softening of the faeces, but the lambs were not severely affected by the coccidia. In lambs given a dose of 10 mg/kg of toltrazuril twice, either on days 10 and 11 after turnout, or on days 10 and 20, the coccidial infection caused a softening af the faeces, including some cases of diarrhoea. Oocyst production due to the initial coccidial infection on pasture was markedly reduced by all treatments with toltrazuril. The reduction was most pronounced after a dose of 20 mg/kg. Lambs treated with single doses of 15 or 20 mg/kg of toltrazuril had a better weight gain than the untreated controls in 2 of the trials. Lambs treated with toltrazuril on day 10 after turnout were partially resistant to the coccidial reinfection acquired immediately after treatment, and they had a similar level of immunity as the untreated controls to the subsequent reinfection on pasture.     

The use of toltrazuril for the prevention of coccidiosis in piglets before weaning

SUMMARY: To determine the efficacy of toltrazuril as a prophylactic treatment for coccidiosis in piglets caused by Isospora suis (I suis) , a single 1.0 mL dose of toltrazuril was administered orally to 1056 piglets between 3 and 6 days of age, in 5 piggeries. Prophylactic treatment of piglets reduced the occurrence of coccidiosis in litters from 71% to 22%. The number of antibacterial treatments given and the number of piglets affected per litter were also significantly reduced, resulting in some improvement in growth rates to weaning. The severity of diarrhoea was significantly reduced, as was the amount of oocyst excretion. The number of days that piglets excreted oocysts in the faeces was reduced from 4.9 days to 2.5 days. The detection of J suis in piglets with diarrhoea was reduced from 84% in the untreated piglets to 6% in the piglets given the prophylactic treatment.     

Effects of the GnRH antagonist acyline on the testis of the domestic cat (Felis catus)

The aim of this study was to describe the effects of a single dose of the gonadotrophin releasing hormone (GnRH) antagonist acyline on testicular characteristics of the domestic cat. Twelve mature cats were orchidectomised unilaterally (right testis) on Day -7 (n=7) or Day 15 (n=5). On Day 0, 330 μg/kg acyline was administered s.c. to all the animals. Left orchidectomy was carried out on Day 15 (n=2), Day 30 (n=4) and Day 60 (n=6). Sperm were recovered from the epididymis and the testes were evaluated grossly, histologically and immunohistochemically. Significant differences (P<0.05) were found between days for epididymal sperm motility, vigor, abnormal morphology, germinal epithelium height, spermatocytes, spermatids, spermatozoa, lumen and cellular debris. Conversely, no significant differences were found for gross testicular and tubular characteristics, spermatogonia, Sertoli and Leydig cells and intertubular compartments. It was concluded that a single dose of acyline reversibly impaired spermiogenesis, spermatocytogenesis and sperm motility for 2 weeks.     

Detection, quantifications and pharmacokinetics of toltrazuril sulfone (Ponazuril®) in cattle

Toltrazuril sulfone (Ponazuril^®) is a triazine-based anti-protozoal agent with highly specific actions against apicomplexan group of organisms, which are undergoing intensive investigation. Toltrazuril sulfone may have clinical application in the treatment of Neospora. caninum and other protozoal infections in cattle. To evaluate absorption, distribution, and elimination characteristics of toltrazuril sulfone in cattle, a sensitive validated quantitative high-pressure liquid chromatography method for toltrazuril sulfone in bovine biological fluids was developed. After a single oral dose of toltrazuril sulfone at 5 mg/kg (as 150 mg/g of Marquis^®; Bayer HealthCare, Shawnee Mission, KS, USA), samples from six cows showed good plasma concentrations of toltrazuril sulfone, which peaked at 4821 ng/mL ± 916 (SD) at 48 h postadministration. Thereafter, plasma concentration declined to 1950 ng/mL ± 184 (SD) at 192 h after administration with an average plasma elimination half-life of ∼58 h. Following oral dose of toltrazuril sulfone, the observed peak plasma concentrations were in relatively close agreement ranging from the lowest 3925 ng/mL to the highest of 6285 ng/mL with the mean peak plasma concentration being 4821 ng/mL. This study shows that toltrazuril sulfone is relatively well absorbed after oral dose in cattle. These results are therefore entirely consistent with and support the reported clinical efficacy of toltrazuril sulfone in the treatment of experimentally induced clinical cases of N. caninum and other protozoal-mediated bovine diseases.     

几种抗球虫药物对獭兔球虫感染和幼兔生长性能的影响

在福建省福安市某獭兔场共检测出16种艾美耳球虫,均为混合感染。选取210只断乳1周、球虫卵囊阳性的獭兔幼兔,随机分为7组,进行抗球虫药物试验。在抗球虫药物试验组中,2.5%妥曲珠利组的獭兔幼兔按有效剂量30 mg/kg体重、5%肠球双效(盐酸氨丙啉+胺喹噁啉钠)组和5%优球乐(癸氧喹酯)组按有效剂量50 mg/kg体重灌服,分为1次用药组和2次用药组;对照组灌服等量生理盐水。结果显示,妥曲珠利组獭兔幼兔用药后第1周,粪便中的球虫卵囊数(OPG值)比用药前减少78.39%,也显著低于其他药物组和对照组(P<0.01);用药后第2周,球虫OPG值比用药前减少88.35%,感染率也降至最低;用药后第8周,粪便中球虫OPG值比用药前减少92.99%。至用药后第8周,各试验组与对照组獭兔幼兔球虫OPG值均转归为相对较低水平,感染率则仍保持较高水平;妥曲珠利组獭兔幼兔死亡率显著低于对照组(P<0.05),其他组间死亡率差异不显著;其平均增重显著高于对照组(P<0.05),料重比则显著低于对照组(P<0.05)、极显著低于5%肠球双效1次用药组(P<0.01)。     

Effects of toltrazuril on the growth of piglets in herds without clinical isosporosis

Isospora suis is a widely prevalent and economically important parasite. The antiprotozoal compound, toltrazuril, was given as a single treatment to piglets without clinical signs in 10 herds. The daily weight gain (DWG) and mortality between the treatment date and weaning was compared on each herd in 10 treated and 10 control litters. The faeces of control litters were examined for oocyst excretion. In six herds, no oocysts were detected and treatment had no effect on DWG or mortality. In four pig herds, oocysts were detected and toltrazuril treatment significantly improved DWG by 25 g (P=0.003). Mortality rate was not affected. The partial economic benefit of toltrazuril treatment in positive herds was 0.20 euros per piglet. A single toltrazuril treatment of piglets in herds without clinical signs of isosporosis but with oocysts detected significantly improved DWG and was considered economically justified on each farm.     

Eprinomectin pour-on for control of Boophilus microplus (Canestrini) ticks (Acari: Ixodidae) on cattle

The efficacy of a commercial pour-on formulation of eprinomectin, a macrocyclic lactone, against experimental infestations of Boophilus microplus (Canestrini) ticks was evaluated in two trials involving 27 Bos taurus calves. The first trial was designed to evaluate the effects of a single treatment at a dose of 0.5 mg/kg of body weight against standard size B. microplus females (4.5-8.0 mm long). A significant reduction in tick numbers (P<0.05, Wilcoxon test) was observed between treated calves as compared to untreated ones from Day 3 (44% efficacy) after treatment to the end of the trial on Day 28 (96.9%), with a peak efficacy of 97.1% on Day 21. In the second trial the effect of eprinomectin on standard size tick numbers, engorgement weight and fertility of female ticks from calves with a single treatment dose of 1 mg/kg on Day 0 and calves treated twice at a dose of 0.5 mg/kg on Days 0 and 4 was evaluated. An efficacy >93% was obtained from Day 2 to Day 28 after treatment in calves treated twice at 0.5 mg/kg, and to the end of the trial (Day 35) in calves treated once with 1 mg/kg. The 1mg/kg treatment provided >98% residual efficacy for at least 7 days. During the first part of the second trial the efficacy of eprinomectin resulted from a dramatic adverse effect on engorgement weight and fertility of female ticks, with 100% control on Day 5 (dosage of 1 mg/kg) and on Days 6 and 7 (two doses of 0.5 mg/kg). Following Day 7, most of the effect was due to reduction in the number of standard size female ticks.     

Pharmacokinetics of toltrazuril and its metabolites in pregnant and nonpregnant ewes and determination of their concentrations in milk,allantoic fluid,and newborn plasma

The objectives of this study were to determine the pharmacokinetics of toltrazuril and its metabolites in pregnant and nonpregnant ewes following a single oral dose and to determine the plasma concentrations of these compounds in milk, allantoic fluid, and newborn plasma. Eighteen healthy ewes were randomly divided into three groups (n = 6 each): pregnant ewes at 12–13 weeks of gestation (group A), nonpregnant ewes (group B), and pregnant ewes at 1–2 weeks before expected lambing date (group C). Ewes in all groups received a single oral dose of toltrazuril at 20 mg/kg body weight. In groups A and B, blood samples were collected at 1, 3, 5, 7, 9, 12, 15, 18 hr, every 6 hr to day 3, every 12 hr to day 7 and thereafter every 24 hr to day 14 post-toltrazuril administration. In group C, parturition was induced 24–36 hr after toltrazuril administration then milk, allantoic fluid, and newborn plasma samples were collected immediately after birth. Drug metabolites were assayed using ultra high-performance liquid chromatography–ultraviolet detection method (UHPLC-UV). The maximum concentration (C_max), area under the plasma concentration-time curve (AUC_0–t), AUC to 24 and 48 hr (AUC_0–24), and (AUC_0–48) were significantly higher in pregnant ewes. Longer apparent half-life (T_1/2), significantly higher apparent volume of distribution (Vd/F) and total clearance (Cl/F) were observed in nonpregnant ewes. The time to maximum plasma concentration (T_max), mean residence time (MRT) and elimination rate constant (K_el) were similar in both groups. The AUC_0–24 and AUC_0–48 were significantly higher in nonpregnant ewes. The AUC_0–t was significantly higher in pregnant ones. The ratio of plasma toltrazuril concentrations in ewes and toltrazuril concentrations in newborn lambs' plasma, allantoic fluid, and milk were 68%, 2.3%, and 5.3%, respectively. Results of this study showed that toltrazuril is well absorbed after a single oral dose in ewes with widespread distribution in different body tissues.     

Toltrazuril sulfone sodium salt: synthesis, analytical detection, and pharmacokinetics in the horse

Toltrazuril sulfone (ponazuril) is a triazine-based antiprotozoal agent with clinical application in the treatment of equine protozoal myeloencephalomyelitis (EPM). In this study, we synthesized and determined the bioavailability of a sodium salt formulation of toltrazuril sulfone that can be used for the treatment and prophylaxis of EPM in horses. Toltrazuril sulfone sodium salt was rapidly absorbed, with a mean peak plasma concentration of 2400 ± 169 (SEM) ng/mL occurring at 8 h after oral-mucosal dosing and was about 56% bioavailable compared with the i.v. administration of toltrazuril sulfone in dimethylsulfoxide (DMSO). The relative bioavailability of toltrazuril sulfone suspended in water compared with toltrazuril sulfone sodium salt was 46%, indicating approximately 54% less oral bioavailability of this compound suspended in water. In this study, we also investigated whether this salt formulation of toltrazuril sulfone can be used as a feed additive formulation without significant reduction in oral bioavailability. Our results indicated that toltrazuril sulfone sodium salt is relatively well absorbed when administered with feed with a mean oral bioavailability of 52%. Based on these data, repeated oral administration of toltrazuril sulfone sodium salt with or without feed will yield effective plasma and cerebrospinal fluid (CSF) concentrations of toltrazuril sulfone for the treatment and prophylaxis of EPM and other protozoal diseases of horses and other species. As such, toltrazuril sulfone sodium salt has the potential to be used as feed additive formulations for both the treatment and prophylaxis of EPM and various other apicomplexan diseases.     

Efficacy of toltrazuril against intestinal and hepatic coccidiosis in rabbits

The anticoccidial effect of toltrazuril (Bay Vi 9142) against Eimeria flavescens, E. intestinalis, E. magna, E. perforans and E. stiedai was tested in experimentally-infected rabbits. Continuous administration of 10-15 p.p.m. of the drug in the drinking water was highly effective in reducing oocyst output of all five species and in preventing clinical signs and macroscopic lesions. Sporulation of excreted oocysts was not affected. After 5 weeks of medication, no negative influence was noted on zootechnic performance of growing healthy rabbits. Medication of rabbits with 25 p.p.m. only during schizogony or gamogony (2 days of treatment, repeated after 5 days) quickly reduced clinical signs and oocyst output. When administered during late schizogony or gamogony, toltrazuril allowed development of immunity against reinfection with homologous species.     

Synthesis and detection of toltrazuril sulfone and its pharmacokinetics in horses following administration in dimethylsulfoxide

Triazine-based antiprotozoal agents are known for their lipophylic characteristics and may therefore be expected to be well absorbed following oral administration. However, although an increase in lipid solubility generally increases the absorption of chemicals, extremely lipid-soluble chemicals may dissolve poorly in gastrointestinal (GI) fluids, and their corresponding absorption and bioavailability would be low. Also, if the compound is administered in solid form and is relatively insoluble in GI fluids, it is likely to have limited contact with the GI mucosa, and therefore, its rate of absorption will be low. Based on the above considerations, we sought a solvent with low or no toxicity that would maintain triazine agents in solution. As the oral route is most preferred for daily drug therapy, such a solvent would allow an increased rate of absorption following oral administration. In present study, it was demonstrated that dimethylsulfoxide (DMSO) increased the oral bioavailability of toltrazuril sulfone (Ponazuril) threefold, relative to oral administrations of toltrazuril sulfone suspended in water. The cross-over study of toltrazuril sulfone formulated in DMSO indicated that the absolute oral bioavailability of toltrazuril sulfone in DMSO is 71%. The high bioavailability of the DMSO-preparation suggests that its daily oral administration will routinely yield effective plasma and cerebral spinal fluid (CSF) concentrations in all horses treated. Also, this improved formulation would allow clinicians to administer loading doses of toltrazuril sulfone in acute cases of Equine Protozoal Myeloencephalitis. Another option would involve administration of toltrazuril sulfone in DMSO mixed with feed (1.23 kg daily dose) meeting the US Food and Drug Administration (FDA) recommendations for the levels of DMSO permissible in pharmaceutical preparations.     

通扬球虫清对鸡球虫病的疗效试验

测定了通扬球虫清对鸡球虫病的疗效,三批试验结果显示,通扬球虫清高剂量组(甲基三嗪酮37.5mg/L饮水浓度,8天)和中剂量1组(甲基三嗪酮25mg/L饮水浓度,8天)的抗球虫指数始终在高效水平;中剂量2组(甲基三嗪酮25mg/L饮水浓度,2天)和通扬独霸组(地克珠利0.5mg/L饮水浓度,8天)的抗球虫指数相近,也属高效抗球虫范围;通扬球虫清低剂量组(甲基三嗪酮12.5mg/L饮水浓度,8天)的抗球虫指数属中等有效水平。综合各项指标及与通扬独霸组的药效相比,通扬球虫清在临床上选用25mg/L饮水浓度,连续饮用2天或2天以上切实可行。     

Coccidiosis control with toltrazuril in conjunction with anticoccidial medicated or nonmedicated feed

A 42-day broiler floor pen study was conducted comparing the anticoccidial efficacy of toltrazuril (Baycox) as a stand alone treatment and as an additional treatment to in-feed anticoccidial programs. Toltrazuril was administered on days 18 and 19 in the drinking water at 7 mg/kg of body weight. The treatments were 125 ppm nicarbazin (days 0-14) to 66 ppm salinomycin (SAL) (days 15-35) with and without toltrazuril, SAL (days 0-35) with and without toltrazuril, nonmedicated (NM) to SAL with toltrazuril, and NM with and without toltrazuril. The controls were NM noninfected and infected. The treatments were replicated in five blocks of eight pens each in a randomized complete block design. All withdrawal feed was nonmedicated. On day 14, birds, except noninfected, were exposed to coccidial oocysts (Eimeria acervulina, Eimeria maxima, and Eimeria tenella) seeded litter. On days 21, 28, 35, and 42, birds and feed were weighed, four birds per pen were coccidial lesion scored, and litter oocyst counts were performed. The coccidial infection in the NM infected treatment caused a significant (P < 0.05) coccidiosis infection. Coccidiosis was moderately controlled in the anticoccidial treatment birds without toltrazuril. Performance in the NM with toltrazuril was equal to or better (P < 0.05) than the anticoccidial programs without toltrazuril. Toltrazuril was equal to the noninfected birds in performance. Toltrazuril most completely eliminated all coccidial lesions and dramatically reduced oocyst shedding. The performance data, lesion scores, and oocyst counts showed that a 2-day treatment with toltrazuril successfully controlled the coccidiosis with no relapse of infection. Toltrazuril can thus be used for supplemental control with in-feed anticoccidials or as a primary anticoccidial with nonmedicated feed.     

Immunohistochemical observations on the initial disorders of the epiphyseal growth plate in rats induced by high dose of vitamin A.

The initial disorders of the epiphyseal growth plate cartilage were immunohistochemically examined in the proximal tibia of rats administered a high dose of vitamin A. Male Wistar rats were given 100,000 IU/100 g body weight/day of vitamin A for administration periods of 1 to 5 days (Day 1 to 5) from 4 weeks after birth or were given deionized water and used as control. They were sacrificed after 5-bromo-2'-deoxyuridine (BrdU) injection on Day 1 to Day 5 to remove the tibiae. The tibiae were processed for immunohistochemical examinations using antibodies against type I, II, X collagens and BrdU. BrdU-incorporated chondrocytes and type X collagen-negative area were reduced since Day 2 and type X collagen-positive area was reduced since Day 4. The cartilage matrix partially lost type II collagen and deposited type I collagen in the epiphyseal growth plate near the periosteum on Day 5. These findings suggest that a high dose of vitamin A initially disturbed the differentiation from resting to proliferating chondrocytes, subsequently inhibited the differentiation from proliferating to hypertrophic chondrocytes, caused the chondrocytes to deviate from the process of normal differentiation, and finally resulted in the deformation of the epiphyseal growth plate.     

Efficacy of toltrazuril and clazuril against experimental infections with Eimeria labbeana and E. columbarum in racing pigeons

The efficacy of toltrazuril in comparison with clazuril on heavy experimental Eimeria labbeana and E. columbarum infections in pigeons was investigated. The minimum required dose of toltrazuril to completely suppress oocyst excretion is 20 mg/kg body weight for 1 day. After treatment with toltrazuril, there was a dose-dependent period during which pigeons remained negative; at a dosage of 35 mg/kg or higher, this period was at least 4 weeks. Clazuril at the recommended dose of 2.5 mg/pigeon resulted in a rapid suppression of oocyst excretion, but oocysts were again observed in the feces 20 days after treatment. The fact that oocysts appeared again in the feces 3 to 5 weeks after treatment can probably be explained only by a persistence of endogenous stages, which are not affected during treatment, rather than by a reinfection. Reinfection of the previously medicated and infected pigeons 30 days after the first infection provided some data on possible induced immunity.     

Safety evaluation of orally administered afoxolaner and milbemycin oxime in eight‐week‐old dogs

The safety profile of afoxolaner (an isoxazoline molecule) when combined with milbemycin oxime (a macrocyclic lactone) was evaluated according to the regulatory requirements when administered six times orally in a soft chewable formulation at a dose of at least 1×, 3×, or 5× the maximum exposure dose in 8‐week‐old Beagle dogs. Thirty‐two healthy puppies (16 males and 16 females) were enrolled and allocated randomly to one of four treatment groups. Three doses were administered at 28‐day intervals (Days 0, 28, and 56), followed by three additional doses administered with 14‐day intervals (Days 84, 98, and 112). The study ended on Day 126. Treatment groups were as follows: Group 1: untreated, sham‐dosed control; Group 2: afoxolaner/milbemycin oxime chews administered at a dose of at least 5 and 1 mg/kg, respectively (1×); Group 3: afoxolaner/milbemycin oxime chews administered at a dose of at least 15 and 3 mg/kg, respectively (3); and Group 4: afoxolaner/milbemycin oxime chews administered at a dose of at least 25 and 5 mg/kg, respectively (5×). All dogs were examined for general health twice a day beginning on Day ‐14. Physical examinations, and blood collections for clinical pathology analysis and afoxolaner and milbemycin oxime plasma concentrations, were performed throughout the study. No afoxolaner/milbemycin oxime treatment‐related changes were observed in growth, physical variables, clinical pathology variables, or tissues examined histologically. No clinically relevant or statistically significant health abnormalities related to the administration of afoxolaner/milbemycin oxime were observed. No signs of macrocyclic lactone sensitivity were observed at any time during the study. Vomiting and diarrhea were observed sporadically across all groups including the controls. Based upon the results of this study, afoxolaner/milbemycin oxime soft chewables were shown to be safe when administered repeatedly at up to 5× the maximum exposure dose in dogs as young as 8 weeks of age.     

Hepatozoonosis in a dog with skeletal involvement and meningoencephalomyelitis

Abstract: A 15‐month‐old, female mongrel dog was presented with a 6‐week history of inappetence, weight loss, and tetraparesis. Physical examination revealed weakness, poor body condition, mild fever, pale mucous membranes, and diffuse muscle atrophy. The right hind limb was painful and edematous, with large ecchymoses. The femur was irregular on palpation and moderate popliteal lymphadenopathy was evident. Results of a CBC showed severe anemia with mild regeneration, an inflammatory leukogram with 90% of neutrophils parasitized by Hepatozoon sp. gamonts, and moderate thrombocytopenia. A bone marrow aspirate had myeloid hyperplasia and contained a few extracellular Hepatozoon meronts and a few intracellular gamonts within neutrophils. Serum chemistry abnormalities included hypoalbuminemia, hyperglobulinemia, hypoglycemia, hypercalcemia, hyperphosphatemia, and elevated alkaline phosphatase activity. Radiologic findings of the right femur included periosteal bone proliferation and lesions compatible with osteomyelitis. A fine needle aspirate specimen from the bone lesion had neutrophilic inflammation; 36% of the neutrophils contained Hepatozoon gamonts. Results of cerebrospinal fluid analysis included a protein concentration of 37 mg/dL and marked mononuclear pleocytosis (243 cell/μL) with a predominance of lymphocytes. An ELISA was positive for Hepatozoon canis and PCR results with DNA sequencing confirmed infection with this organism. A diagnosis of hepatozoonosis with skeletal involvement and meningoencephalomyelitis was made. The dog recovered almost completely neurologically and had no gamonts in the blood after 60 days of therapy with imidocarb dipropionate and prednisone. This is an unusual case of canine hepatozoonosis involving neurologic signs and a periosteal reaction more typical of H. americanum infection and rarely reported in dogs infected with H. canis.