Validation of a blood gas device for ionized calcium analysis in Holstein cows.

BACKGROUND: Accurate analysis of ionized calcium (iCa) is critical for the detection of hypocalcemia or subclinical hypocalcemia. The Edan i15 Vet (EDAN) blood gas device has not been validated for iCa in dairy cows. OBJECTIVES: We aimed to validate the EDAN blood gas device against the Gem Premier 3000 (GEM) analyzer by measuring iCa concentrations and evaluating the ability of these measurements, compared with serum total calcium (TCa) concentrations, to diagnose subclinical hypocalcemia. METHODS: iCa concentrations were measured with the EDAN and GEM devices, and serum TCa concentrations were measured with a wet biochemistry method with blood from 125 lactating Holstein cows between calving to day 27 postpartum. RESULTS: Bland-Altman plots showed a mean and total bias of 0.05 and 0.24 mmol/L for the EDAN device, respectively. The intercept did not include zero, but the slope included 1.0 in the Passing-Bablok regression. The sensitivity and specificity (Se/Sp) of the EDAN device were 93/94%, 93/90%, 91/93%, and 85/95% for iCa cut-off values of <1.00, 1.05, 1.10, and 1.15 mmol/L, respectively, as determined with the GEM device. The Se/Sp were 57/82% and 72/80% for EDAN and 57/80% and 72/79% for GEM at serum TCa cut-off points <2.15 and <2.00 mmol/L, respectively. The average iCa concentrations analyzed with the GEM and EDAN devices were 1.04 ± 0.18 and 1.09 ± 0.17, respectively. CONCLUSIONS: The EDAN device did not have satisfactory agreement with GEM and could not be used interchangeably, but it had satisfactory Se/Sp to diagnose subclinical hypocalcemia compared with the GEM-derived iCa cut-off points. Serum TCa concentration cut-off values were not suitable for diagnosing subclinical hypocalcemia because of unsatisfactory Se/Sp compared with iCa concentrations analyzed by the GEM and EDAN devices. The iCa values analyzed using the EDAN and GEM devices were consistent with previously reported data.

In-vivo efficacy of toltrazuril on experimentally induced Toxoplasma gondii tissue cysts in lambs: a novel strategy for prevention of human exposure to meat-borne toxoplasmosis.

The aim of the present study was to investigate in vivo efficacy of toltrazuril on Toxoplasma gondii tissue cysts following induction of chronic toxoplasmosis in 4-week-old lambs (n=27) by inoculation of 1×10(5) T. gondii ME 49 strain oocysts (day 0). Beginning at the 15th day after inoculation, lambs in Group T20 and Group T40 were given toltrazuril orally 2 times, once every week (Baycox 5%, Bayer Animal Health) at a dose of 20 mg/kg and 40 mg/kg, respectively. Positive control (PC) lambs were not given any therapy, and 2 clinically healthy non-infected lambs were used as negative controls (Group NC). Two out of 9 lambs in PC group (oocyst inoculated but non-treated) were killed on toltrazuril treatment days (day 15 and 22) to evaluate the tissue cyst presence in their muscles. On day 90, the remaining 25 lambs were necropsied, and samples from the brain and 11 different muscle groups were collected. The tissues were examined for the presence of tissue cysts by histopathology, immunohistochemistry, nested-PCR and percoll gradient centrifugation. Anti-T. gondii antibodies were screened by IFAT throughout the experiment. The increased T. gondii seropositivity beginning from the 15th day of inoculation remained steady at Day 45 and Day 90 in Groups PC while it was significantly lower at Day 90 in toltrazuril receiving groups. In toltrazuril treated groups, histopathological findings included degenerative changes in the cyst wall, complete macrophage invasion to the cysts, and reduction or removal of the cysts in toto. Four out of 9 lambs (44.4%) in both toltrazuril treated group (Group T20 and T40) did not contain tissue cyst in any examined tissues while all positive control animals had T. gondii tissue cysts at least in one muscle group. The toltrazuril treatment efficacy on the cyst presence was determined as 44.4%. The number of the cysts in the musculature was significantly different between non-treated and toltrazuril treated lambs (X(2)=6.613; p=0.037). For the total number of cysts, the positive control lambs had higher number of cysts compared to both toltrazuril treated lambs (T20 and T40) (X(2)=5.629; p=0.018 and X(2)=5.629; p=0.018, respectively) while there were no differences between Group T20 and Group T40 (X(2)=0.000; p=1.000). According to PCR results, the brain and M. semitendinosus were positive in all 7 control lambs while 12 out of 18 lambs were positive in toltrazuril treated lambs. In conclusion, the results are promising as the toltrazuril treated lambs had markedly less parasite counts compared to those of untreated lambs. Further research should be conducted to reveal if toltrazuril treatment in sheep could be used as a strategy to minimize the cyst exposure of humans through consumption of raw or undercooked mutton.

Efficacy of toltrazuril 5 % suspension against Eimeria bovis and Eimeria zuernii in calves and observations on the associated immunopathology.

16 Calves were each infected with suspensions containing a mixture of approximately 230,000 Eimeria bovis and 70,000 E. zuernii oocysts, which resulted in detection of oocysts in faeces of 12 of 16 calves by day +14 after infection. On day +14 after infection calves were either treated (n = 8) with toltrazuril at 15 mg/kg body weight or with a placebo. Observations were made on the clinical condition, faecal score and liveweight of calves daily from one day post infection (pi) until 24 days pi when all calves were euthanised and examined post mortem. Samples were collected from ileum and colon for histological, immunohistochemical and gene expression studies. The study demonstrated an efficacy of toltrazuril for the treatment of E. bovis and E. zuernii infections in calves reaching 99 % (based on arithmetic mean oocyst counts in faeces) within three days of treatment and remaining at or above this level for six days. Toltrazuril did not have a significant effect on the pattern and extent of immune cellular infiltration in the mucosa of ileum and colon, but the expression of the genes coding IL-2, IL-10 and TNF-α in the ileum and TNF-α in the colon were elevated in calves treated with toltrazuril. Higher levels of oocyst shedding were significantly associated with lower expression of genes coding for IL-2, IL-10 and higher IP-10. It is concluded that toltrazuril is effective for the treatment of coccidiosis due to E. bovis and E. zuernii in calves and enables the development of a normal or enhanced immune response to infection.