the dry mass of the plantago asiatica was purchased from an oriental pharmacy ( iksan , korea ) , was according to the standard as mentioned in korean pharmacopoeia and korean herbal pharmacopoeia , which are the official compendia of standard . the procedure for preparing pae was as follows . the air - dried mass of plantago asiatica ( 100 g ) was cut into pieces and extracted twice with 50% ( v / v ) ethanol ( six times as much as the weight of the dried plants ) for 3 hours at 80. after filtration through a 400-mesh filter cloth , the filtrate was refiltered through filter paper ( whatman , no . 5 ) and concentrated on a rotary evaporator ( eyela , tokyo , japan ) and the concentrated filtrate was evaporated to dryness under vacuum with freezing dryer ( labconco , usafinally , the solid residue was collected , placed in sealed bottles and stored at 20. the extract yield of plantago asiatica with 50% ethanol was 15.55% . plantamajoside , which was used for the standard material of pae composition , was purchased from the natural product bank , institute for korea traditional medical industry ( geong - san , korea ) . plantago asiatica is a member of the plantaginaceae family , is widely distributed in east asia , used in chinese medicine , where its leaf is called plantaginis herba and its seed is plantaginis semen , and has been commercialized as an extract and as a dietary supplement in various forms . a previous report discussed the isolations of plantamajoside , plantagin and aucubin from p. asiatica , as well as the various biological properties of these compounds , including antibacterial , antiallergic , antiinammatory , antioxidative and lipoxygenase inhibitory activity . figure 1 shows the chemical structures of phenylethanoid glycosides ( phgs ) including acteoside and plantamajoside . waters acquity lc system ( waters corp . , milford , usa ) was used for lc system . the column was c18 type acquity uplc beh ( 2.150 mm , 1.7 m , waters corp . , milford , usa ) . a waters nova pack c-18 column ( acquity uplc beh ( 2.150 mm , 1.7 m , waters corp . , the column temperature was set at 30 with a flow rate of mobile phase at 0.6 ml / min ( 0.1% h3po4/acetonnitrile ) . this study was conducted on the 3-day - old chickens ( n=30 ) in the animal facility of center for animal resources development , wonkwang university , korea . animals were acclimatized and kept in an animal facility room with regulated temperature ( 282 ) , humidity ( 505% ) and light / dark cycle ( 12/12 hours ) . the animals were fed commercial post - broiler diet without antibiotics and coccidiostat ( hanil feed co. , yongin , korea ) and tab water ad libitum . all studies were performed in accordance with the guide for animal experimentation by wonkwang university and approved by the institutional animal care and use committee of wonkwang university . those animals were divided with 3 groups ; pae 0.1% treated / infected ( n=10 ) , pae untreated / infected ( n=10 ) and non - infected control ( n=10 ) . chickens were fed a standard diet supplemented with or without pae for 1 week prior to infection with e. tenella ( 10,000 sporulated oocysts per chicken ) . the effects of pae on e. tenella infection were assessed by two parameters , fecal oocysts shedding and body weights gain . oocysts of e. tenella were cleaned by flotation on 5.25% sodium hypochlorite and washed three times with phosphate buffered saline . e. tenella was provided kindly by professor wongi min at gyeongsang national university in korea . chickens were treated orally by gavages using a 24 gauge , mouse stainless steel animal feeding tube ( popper & sons , inc . the oral infectious dose of has been approximated 10 oocysts of e. tenella in 1 ml of saline . the control chickens ( n=10 ) received saline through the same route . during the study period body weights were individually measured for 2 weeks before infection and for 10 days post - infection . the fecal samples were analyzed for the presence of coccidial oocysts using a standard fecal flotation technique . briefly , 5 ml from each sample was pelleted by centrifugation at 1500g for 5 min . the resulting pellet was resuspended in saturated sodium chloride ( aqueous ) , passed through a 1 mm mesh size sieve to remove coarse fecal debris . the resulting filtrate was used in a standard gravity vial fecal flotation using 22 mm22 mm coverslips . after flotation , the coverslip was mounted on a slide and examined in its entirety for the presence of coccidial oocysts . total number of oocysts was calculated using the following formula : [ total number of oocysts = oocyst countdilution factor(fecal sample volume / counting chamber volume)/number of birds per cage ] . differences in mean oocysts production and mean weight gain between the 4 groups were tested by using one - way analysis of variance ( anova ; graphpad instat ; graphpad software inc . , the effects of pae on e. tenella infection were assessed by two parameters , fecal oocysts shedding and body weight gain . pae - fed chickens produced significantly reduced fecal oocysts ( p<0.05 ) when compared to the e. tenella - infected group fed standard diet . also , pae - based diet , improved body weight loss caused by e. tenella infection . the results showed that , compared to untreated controls , chickens treated with pae had significantly decreased fecal oocysts shedding and showed strong anticoccidial activities ( p<0.01 ) . as shown in table 1 , oocyst shedding was significantly higher in the inoculated chickens than in the control chickens ( p<0.05 ) . the number of fecal oocysts shed was highest on day 7 post - inoculation ( table 1 ) . moreover , body weight gain was lesser in the animals in the inoculated group than in the animals in the control group ( table 2 ) . coccidiosis of domestic fowl is a worldwide disease caused by obligatory intracellular protozoa of the genus eimeria . e. tenella is important pathogen causing avian coccidiosis in laboratory avian animals and known to affect influencing experimental results obtained with contaminated animals . the disease is characterized by enteric lesions of variable extent and severity , reducing the absorptive function of the intestinal mucosa , thus leading to weight loss , diarrhea , poorer feed conversion and a higher mortality in the affected flocks . the results of this study showed that pae had a strong anticoccidial effect on e. tenella . the major bioactive constitute of p. asiatica are phenylethanoid glycosides ( phgs ) including acteoside and plantamajoside as shown in scheme 1 . phgs from many plants have been extensively studied for their various biological functions such as anti - hepatotoxic , anti - inflammatory , antinociceptive activities , improving sexual function   and antioxidant activity . the plantaginin , plantamajoside , acetoside and 6-hydroxyluteolin 7-glucoside isolated from p. asiatica were the chief compounds   and they had various biological activities . the plantamajoside , in particular , was reported to show antibacterial activity against staphylococcus aureus , inhibition activity against camp phosphodiesterase and 5-lipoxygenase , and antioxidant activity on adp+ nadph - induced lipid peroxidation in rat liver microsome . also , the plantamajoside is used as a marker compound in chemotaxonomic studies , as are other members of the dihydroxyphenylethyl glycoside family . theses phenylethanoid glycosides - included bioactive structures could be suggested a plausible explanation of the anticoccidial properties in this study . in this study , the pae - fed chickens produced significantly reduced fecal oocysts ( p<0.05 ) when compared to the e. tenella - infected group fed standard diet . also , pae - based diet , improved body weights loss caused by e. tenella infection .
anticoccidial effects of the plantago asiatica extract ( pae ) were evaluated in chickens following oral infection with eimeria ( e. ) tenella . 
 this study was conducted on the 3-day - old chickens ( n=30 ) . 
 those animals were divided with 3 groups ; pae 0.1% treated / infected ( n=10 ) , pae untreated / infected ( n=10 ) and non - infected control ( n=10 ) . 
 chickens were fed a standard diet supplemented with or without pae for 1 week prior to infection with e. tenella ( 10,000 sporulated oocysts per chicken ) . 
 the effects of pae on e. tenella infection were assessed by two parameters ; fecal oocysts shedding and body weights gain . 
 the pae - fed chickens produced significantly reduced fecal oocysts ( p<0.05 ) when compared to the e. tenella - infected group fed standard diet . 
 also , pae - based diet , improved body weight loss caused by e. tenella infection . 
 our data demonstrated that pae had remarkable anticoccidial activities against e. tenella . 
 this finding might have implications for the development of anticoccidial drug . 
 this study is the first to demonstrate anticoccidial effect of pae on eimeria parasites .