Background: Coccidiosis is one of the most common diseases that hinder the raising of rabbits. The disease causes symptoms such as diarrhea and loss of appetite. Acute infection causes bloody feces and negatively affects fertility. Diarrhea may be bloody due to the intestinal epithelium withering off as many oocysts and merozoites burst out of the cells. This leads to the deaths of rabbits resulting in huge economic losses for commercial rabbit systems. 
 
Methods: The study aims to determine the optimal effective concentration of bile collected from healthy sheep gall bladder inhibiting in vitro sporulation and viability of E. magna and E. exigua oocysts and sporozoites isolated from rabbit intestine. Un-sporulated oocysts were exposed (1&times;102) to six treatments: 2.5% potassium dichromate solution as the non-treated control, four concentrations of bile (12.5%, 25%, 50% and 100%) and Toltrazuril 25 mg/ml as traditional medicine for anti-oocyst activities. In addition, 250, 500, 750 and 1000 mg/mL concentrations of bile were evaluated for effect on sporozoite viability. The oocysts were examined after 25, 50, 75 and 100 hours and sporozoites after 12 and 24 hours of treatment. 
 
Result: GC-Mass analysis showed that sheep bile contains 12 biologically active compounds. In addition, quantitative analysis revealed total phenols (50.65&plusmn;0.5) and total flavonoids (9.39&plusmn;0.4). After 100 hours of exposure, sheep bile was able to prevent sporulation of E. magna and E. exigua oocysts at 100% and 50% concentrations and the rate of inhibition was about 96% and 79%, respectively. Additionally, bile exhibited the least discouraging of 11% at a dose of 125 mg/mL and the highest suppression of 95% of E. magna and E. exigua sporozoites viable at 1000 mg/ml. Generally, the inhibition rate typically increased with longer incubation times and greater doses. Findings have scientifically validated the use of bile in fighting against coccidiosis. Additional studies are required to separate the active compounds found in bile and their mechanisms of action and potential applications in rabbit farms.

In vitro Studies on Anticoccidial Effects of Healthy Sheep Bile against Eimeria magna and Eimeria exigua Oocysts and Sporozoites Isolated from Domestic Rabbits

<div>Background: A novel, rapid and specific multiplex polymerase chain reaction was developed to diagnose hemo-parasitic infection in bovine blood co-infected with three of the most common hemo-parasites.&nbsp; 
 
Methods: The diagnostic process relied on the detection of the three different bovine hemoparasites isolated from red blood cells (RBCs) of cattle (N=30) by conventional Giemsa stained blood smear (GSBS) and confirmed by multiplex PCR. The multiplex PCR system was used to diagnose GSBS positive blood samples (N=12) found infected or co-infected with hemoparasites. The designed multiplex primer sets was attempted to amplify 205, 313 and 422 bp fragments of apocytochrome b, sporozoite and macroschizont 2 (spm2) and 16S rRNA gene for Babesia bigemina, Theileria annulata and Anaplasma marginale, respectively. 
 
Result: This multiplex PCR was sensitive with the ability to detect the presence of 150 ng of genomic DNA. The primers used in this multiplex PCR also showed highly specific amplification of specific gene fragments of each respective parasite. Comparing the two detection methods revealed that 58.33% of specimens showed concordant diagnoses with both techniques. The specificity, positive predictive value and kappa coefficient of the agreement was highest for diagnosis of B. bigemina and lowest for A. marginale. The overall Kappa coefficient for diagnosis based on GSBS for multiple pathogens compared to multiplex PCR was 0.56, slightly behind the threshold of 0.6 of agreement. Therefore, confirmation should always be based on PCR to rule out false positives due to differences in subjective observations, stain particles and false negatives due to low parasitemia. The simplicity and rapidity of this specific multiplex PCR method make it suitable for large-scale epidemiological studies and follow-up of drug treatments.</div>

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In vitro Studies on Anticoccidial Effects of Healthy Sheep Bile against Eimeria magna and Eimeria exigua Oocysts and Sporozoites Isolated from Domestic Rabbits

Development of Novel Multiplex PCR for Diagnosis of Co-infected Hemo-parasites in Cattle