Figure 1: Percent lysis of E. tenella oocysts at different contact times with 3% and 4% dilution of WSD
Figure 2: Percent infectivity of sporulated oocysts of E. tenella, contacted with 3% dilution of WSD for 120 minutes, before challenge and one week after

Organism

Interference with Organic matter (1% skim milk)

Mean % reduction in bacterial count or HA titer of viruses


at different contact times (min) with 3 % WSD

5 min                      15 min                             30  min                      60 min

E. coli

+

_

100

100

100

100

100

100

100

100

Streptococcus spp.

+

_

96.6

100

100

100

100

100

100

100

Salmonella Enteritidis

+

_

100

100

100

100

100

100

100

100

Proteus mirabilis

+

_

100

100

100

100

100

100

100

100

NewCastle Disease Virus

+

_

100

100

100

100

100

100

100

100

Avian Influenza (H9N2)

+

_

100

100

100

100

100

100

100

100

Table 1: Mean % reduction in bacterial count or HA titer of viruses at different contact times (minutes) with 3% dilution of WSD

Contact time (min)

Percent EOBWE Dilutions (V/V)

0.005                             0.010                         0.020                            0.100

30

 

 

X1

X

60

 

 

 

 

90

 

 

 

 

120

 

X

 

 

180

X

 

 

 

1 X designates the dilution of EOBWE and the contact time resulting in maximum lysis of the oocysts
Table 2: Impact of EOBWE dilution and contact time on obtaining maximum lysis of equivalent number of pooled oocysts of 8 Eimeria spp

Presence of
1 % skim milk

Average % reduction in skim milk after different
contact time in minutes with EOBWE

Organism

5 min

15 min

30 min

60 min

+

15.0

12.6

4.0

26.4

E. coli

-

35.1

47.3

73.5

78.4

E. coli

+

6.1

6.9

6.2

14.0

Strep. spp.

-

39.2

16.2

37.2

39.0

Strep. spp.

+

85.0

87.0

100.0

100.0

Salmonella Enteritidis

-

90.6

94.1

100.0

100.0

Salmonella
Enteritidis

+

50.5

65.6

54.3

66.9

Proteus mirabilis

-

93.5

97.3

99.4

96.1

Proteus mirabilis

+

39.0

100.0

100.0

100.0

Newcastle Disease virus

-

ND

100.0

100.0

100.0

Newcastle Disease virus

+

100.0

100.0

100.0

100.0

Avian Influenza (H9N2)

-

100.0

100.0

100.0

100.0

Avian Influenza (H9N2)

Table 3: Average % reduction in organisms after 4 different contact times (min) with EOBW

Challenge at age (d)

Mean weight increase of broilers in the 4 treatments at end of 6 days post each challenge

       G1                               G2                                               G3                                                 G4                              SEM2

14

79.2

87.0

21.5

82.6

 

21

45.7

49.3

17.7

47.5

 

28

55.2

57.7

19.4

52.4

 

35

32.4

37.2

15.6

35.7

 

Mean of Means

53.1a

57.8b

18.6c

54.6a

1.2

1The four treatments were: G1 (Unchallenged – Untreated with EOBWE), G2 (Unchallenged – Treated with EOBWE), G3 (Challenged –Untreated with EOBWE), and G4 (Challenged-Treated with EOBWE).
2SEM is Standard Error of Means
aMeans of Means in the last row followed by different alphabet superscripts are significantly different at P<0.05
Table 4: Percent weight increase in broilers of the 4 treatments1 (G1 – G4) during the 6 days post each challenge that was administered at different ages

Age at challenge age (days)

Feed conversion ratio of broilers in Treatments G1 – G4

G1

G2

G3

G4

SEM2

14

1.68

1.58

2.50

1.65

 

 

 

0.16

21

1.41

1.40

2.10

1.45

28

1.47

1.42

2.95

1.44

35

1.54

1.43

2.17

1.47

Mean feed conversion

1.53a

1.46a

2.43b

1.50a

1The four treatments were: G1 (Unchallenged – Untreated with EOBWE), G2 (Unchallenged – Treated with EOBWE), G3 (Challenged –Untreated with EOBWE), and G4 (Challenged-Treated with EOBWE).
2SEM is Standard Error of Means
a-bMeans in a row followed by different alphabetic superscripts are significantly different (P<0.05)
Table 5: Feed conversion ratio of the four differently treated broilers (Treatments1 G1 – G4) during the 6 days post each challenge that was administered at different ages

Age at challenge age (days)

Mortality of broilers in Treatments G1 – G4

G1

G2

G3

G4

SEM2

14

0.0

0.0

30.0

10.0

 

 

 

1.7

21

0.0

0.0

20.0

0.0

28

0.0

0.0

20.0

0.0

35

0.0

0.0

10.0

0.0

Mean % Mortality

0.0a

0.0a

20.0b

2.5a

1The four treatments were: G1 (Unchallenged – Untreated with EOBWE), G2 (Unchallenged – Treated with EOBWE), G3 (Challenged –Untreated with EOBWE), and G4 (Challenged-Treated with EOBWE)
2SEM is Standard Error of Means
a-bMeans in a row followed by different alphabetic superscripts are significantly different (P<0.05)
Table 6: Mortality of broilers in the four Treatments1 (G1 – G4) during the 6 days post each challenge that was administered at different ages

Age at challenge age (days)

Intestinal Organ

Mean gross lesion score in intestinal organs of broilers (Treatments G1 – G4)

G1

G2

G3

G4

SEM3

14

Duodenum
Jejunum
Ileum
Cecum

0.1a
0.0a
0.0a
0.0a

0.0a
0.0a
0.0a
0.0a

2.4b
2.5b
1.7b
1.9b

0.1a
0.1a
0.2a
0.1a

0.13
0.13
0.12
0.10

21

Duodenum
Jejunum
Ileum
Cecum

0.2a
0.1a
0.0a
0.1a

0.1a
0.0a
0.0a
0.0a

2.4b
1.9b
1.5b
1.5b

0.1a
0.0a
0.1a
0.0a

0.12
0.10
0.11
0.09

28

Duodenum
Jejunum
Ileum
Cecum

0.2a
0.0a
0.1a
0.0a

0.1a
0.0a
0.1a
0.0a

2.1b
1.6b
2.5b
2.5b

0.2a
0.1a
0.0a
0.2a

0.14
0.10
0.13
0.10

35

Duodenum
Jejunum
Ileum
Cecum

0.1a
0.1a
0.0a
0.0a

0.2a
0.0a
0.0a
0.0a

2.4b
2.4b
1.7b
1.5b

0.1a
0.0a
0.0a
0.1a

0.13
0.14
0.10
0.10


1Gross lesion in the intestinal organs are given scores of 0 to 4; the score 0 is given to absence of inflammation, while score 1 indicates a mild inflammation, and score 4 indicates sloughing of the epithelial layer with hemorrhage. The scores of 2 and 4 are given to lesions of medium inflammation and to slight sloughing, respectively.
2The four treatments were: G1 (Unchallenged – Untreated with EOBWE), G2 (Unchallenged – Treated with EOBWE), G3 (Challenged –Untreated with EOBWE), and G4 (Challenged-Treated with EOBWE).
3SEM is Standard Error of Means
a-bMeans in a row followed by different alphabetic superscripts are significantly different (P<0.05)

Table 7: Mean gross lesion score1 in intestinal organs of broilers in Treatments2 G1 – G4 at 6 days post each challenge that was administered at different ages

Age at challenge age (days)

Intestinal Organ

Mean Log10 of Oocyst count per gram of intestinal organ

G1

G2

G3

G4

SEM2

14

Duodenum
Jejunum
Ileum
Cecum

0.0a
0.0a
0.0a
0.0a

0.0a
0.0a
0.0a
0.0a

3.7b
5.2b
5.7b
6.3b

1.1c
1.4c
0.6c
2.2c

0.02
0.01
0.03
0.01

21

Duodenum
Jejunum
Ileum
Cecum

0.0a
0.0a
0.0a
0.0a

0.0a
0.0a
0.0a
0.0a

4.1b
5.4b
4.5b
6.7b

1.2c
1.3c
1.1c
2.4c

0.05
0.03
0.03
0.01

28

Duodenum
Jejunum
Ileum
Cecum

0.0a
0.0a
0.0a
0.0a

0.0a
0.0a
0.0a
0.0a

3.1b
5.4b
5.2b
6.7b

1.2c
1.1c
1.1c
2.3c

0.01
0.03
0.01
0.03

35

Duodenum
Jejunum
Ileum
Cecum

0.0a
0.0a
0.0a
0.0a

0.0a
0.0a
0.0a
0.0a

4.3b
5.2b
5.4b
6.2b

1.2c
1.3c
1.1c
2.3c

0.03
0.01
0.02
0.02

1The four treatments were: G1 (Unchallenged – Untreated with EOBWE), G2 (Unchallenged – Treated with EOBWE), G3 (Challenged –Untreated with EOBWE), and G4 (Challenged-Treated with EOBWE).
2SEM is Standard Error of Means
a-cMeans in a row followed by different alphabetic superscripts are significantly different (P<0.05)
Table 8: Mean Log10 of Oocyst count per gram of intestinal organ of broilers in Treatments1 G1 – G4 at 6 days post each challenge that was administered at different ages

Treatment

Challenge

Coccidiostat

Mean Oocyst output/g dropping of two Eimeria spp.
_________________________________________________________
E. acervulina                                                     E. maxima

1

+

-

3.7E+06a

4.0E+04a

2

+

Maxiban

7.7E+05b

0.0b

3

+

EOBWE

2.1E+06a

0.0b

        a,b Means in a column followed by different alphabetic superscripts are significantly different (P<0.05)
Table 9: Mean oocyst output of E. acervulina and E. maxima after 6 days of challenge in broilers that are differently administered (ad libitum) the Maxiban in feed or EOBWE in drinking water

Treatment

Challenge

Coccidiostat

Mean body Weight in Grams

1

+

-

1000a

2

+

Maxiban

1100b

3

+

EOBWE

1100b

a-bMeans in a column followed by similar alphabet superscript are insignificantly different (P>0.05)
Table 10: Mean body weight post 6 days of challenge with Eimeria spp. in broilers that are differently administered (ad libitum) the Maxiban in feed or EOBWE in drinking water

Treatment

Challenge

Coccidiostat dilution or inclusion

Mean Oocyst output/g dropping at two ages (days)

28                                            35

 

1

 

+

 

-

 

(3.2x103)a

 

(4.1x103)a

2

+

EOBWE 0.005%

(0.36x102)b

(1.27x10)b

3

+

EOBWE 0.010%

(0.77x10)c

(0.15x10)c

4

+

EOBWE 0.025%

(0.81x10)c

(0.15x10)c

5

+

Salinomycin 66mg/Kg feed

(0.0)c

(0.0)c

1The EOBWE was administered ad libitum in drinking water at the intermittent age of 4-6, 10-12, 18-20, 24-26, and 31-33 days.
a-cMeans in a column followed by different alphabetic superscripts are significantly different (P<0.05)

Table 11: Oocyst output at different ages of broilers that were exposed to contaminated floors with 4x105 sporulated oocysts/meter squareof Eimeria spp organisms at day 1 and treated intermittently1 with different dilutions of EOBWE in drinking water or Salinomycin in feed

Treatment

Challenge

Coccidiostat dilution or inclusion

Average body weight  (g)

Feed Conversion Ratio

1

+

-

1552.3a

1.745a

2

+

EOBWE (0.005%)

1626.0a,b

1.555b

3

+

EOBWE (0.0125%)

1690.5b,c

1.526b,c

4

+

EOBWE (0.025%)

1710.3b,c

1.503c

5

+

Salinomycin 66mg/Kg feed

1728.7c

1.502c

1The performance was measured by the parameter of live body weight and feed conversion ratio at 35 days of age
2EOBWE was administered ad libitum in drinking water at the intermittent age of 4-6, 10-12, 18-20, 24-26, and 31-33 days.
a-cMeans in a column followed by different alphabetic superscripts are significantly different (P<0.05)

Table 12: Performance1 of broilers exposed to contaminated floors with Eimeria spp organisms at day 1 and treated intermittently with EOBWE in drinking water2 or Salinomycin in feed

Treatment

disinfectant

coccidiostat

Average oocyst output per gram of dropping at different ages (days)

14                   21                   28                   35

1

WSD2

EOBWE3

13,502a

25,207a

15,520a

6,983a

2

WSD

Maxiban

8,506a

18,256a

13,829a

7,028a

3

Chlorine4

EOBWE

98,660b

104,560b

63,135b

16,097b

4

Chlorine

Maxiban

98,660b

91,212b

62,939b

16,326b

1Total sporulated oocysts of 4x105/m2 of equivalent number of oocysts of 8 Eimeria spp. (E. acervulina, E. brunetti, E. hagani, E. maxima, E. mivati, E. necatrix, E. praecox, and E. tenella)
2WSD stands for invented Wide Spectrum Disinfectant, diluted to 3 %, and applied at a rate of 400 ml/m2
3EOBWE stands for invented Essential Oil Blend in Water Extract that is intermittently administered in drinking water for 3 days/week.
4Chlorine was applied according to manufacturer’s instructions.
a-bMeans in a column followed by different alphabetic superscripts are significantly different (P<0.05)
Table 13: Mean Oocysts output recovered from duplicate pens per treatment of broilers exposed at day 1 to floors contaminated by sporulated oocysts of 8 Eimeria spp1followed by decontamination of pens by different disinfectants, and treating the reared birds by different coccidiostats

Treatments

Disinfectant

Coccidiostat

Means of three Production parameters

Feed Conversion    Live body weight (g)         % Mortality
Ratio

1

WSD2

EOBWE3

1.72a

1798a

4.0a

2

WSD

Maxiban

1.93b

1531b

4.5a

3

Chlorine4

EOBWE

1.85b

1638c

6.5b

4

Chlorine

Maxiban

2.03c

1605c

7.0b

1Total sporulated oocysts of 4x105/m2 in equivalent number of oocysts of 8 Eimeria spp. (E. acervulina, E. brunetti, E. hagani, E. maxima, E. mivati, E. necatrix, E. praecox, and E. tenella)
1WSD stands for invented Wide Spectrum Disinfectant, diluted to 3 %, and applied at a rate of 400 ml/m2
2EOBWE stands for invented Essential Oil Blend in Water Extract that is intermittently administered in drinking water for 3 days/week
4Chlorine was applied according to manufacturer’s instructions
a-c Means in a column followed by different alphabetic superscripts are significantly different (P<0.05)
Table 14: Performance of the differently treated broilers with dual interception by disinfectants for decontamination of oocysts on surfaces of pens1 and by administering different coccidiostats