The Effects of Commercially Available Syzygium aromaticum, Anethum graveolens, Lactobacillus acidophilus LB, and Zinc as Alternatives Therapy in Experimental Mice Challenged with Cryptosporidium parvum

Figures & data

Table 1 Primers Sets Used for nPCR Targeting COWP Gene

Primers	Sequences	Expected Product Size (bp)	Annealing Temperature (°C)
1st PCR (E-COWP = Extended COWP)^Citation73
BCOWPF	5ʹ-ACCGCTTCTCAACAACCATCTTGTCCTC-3’	769	65
BCOWPR	5ʹ-CGCACCTGTTCCCACTCAATGTAAACCC-3’
2nd PCR (N-COWP = Nested COWP)^Citation74
Cry-15	5ʹ-GTA GAT AAT GGA AGA GAT TGT G-3’	553	54
Cry-9	5ʹ-GGA CTG AAA TAC AGG CAT TAT CTT G-3’

Table 2 Treatment Type and Dose Were Given to the Mice of T Subgroups

Groups	Treatment	Treatment Dose	Time of Treatment in Relation to Infection	Duration of Treatment
T1	No	–	–	–
T2	NTZ	200 mg/kg/day, twice daily	7th PID	Eight consecutive days
T3	Clove oil	33 mg/kg, once daily
T4	Dill seed oil	20 μL, once daily
T5	Lactobacillus acidophilus LB	5×10^8 dissolved in 200 µL distilled water, once daily
T6	Zinc	5 mg in 500 µL syrup, once daily

Table 3 Rate of Infection Throughout the Experiment

Group	No. Samples	−ve Samples	+ve Samples	Percentage of +ve Samples
Control	46	46	0	0
T1	47	12	35	74.46
T2	39	24	15	38.5**
T3	49	15	34	69.38
T4	38	19	19	50**
T5	42	20	22	52.4**
T6	40	22	18	45**
Total	301	158	143	47.5**

Note: Values in the last column followed by **are highly significant (P-value ≤0.001).

Table 4 Comparison of Means Oocysts/gm Stool Detected in All Groups at the End of the Experiment and Reduction Rate of Each Treatment

Groups	Mean of Oocysts/gm Stool	Std. Deviation	Reduction Rate
T1	4970^a	±135.29	–^a
T2	420	±4.38	91.55 ^b
T3	1260	±10.57.	74.65^bc
T4	210	±1.96	95.77 ^b
T5	210.00	±1.80	95.77 ^b
T6	98.00	±1.76	98.03 ^b

Note: Oocysts discharge reduction rates among different groups differ significantly (P <0.05) when they are not sharing any common superscript letter (a–c).

Table 5 Body Weights in All Groups Throughout the Experiment Compared to the Control Group

Groups	Mean	Std. Deviation	P-value
C	30.2457	±0.21547	–
T1	27.8214	±0.24862	**
T2	29.1000	±0.15191	**
T3	27.9286	±0.22678	**
T4	28.5571	±0.08516	*
T5	29.0643	±.017305	**
T6	30.8071	±0.12157	*

Note: Values in each column followed by * means P-value is significant (P-value ≤ 0.05) and that followed by ** are highly significant (P-value ≤0.001).

Table 6 IL10 Difference Among Groups Compared to Control

Group	No. of Samples	Mean (pg/mL)	Std. Deviation	P value
C	6	1749.9	23.53	–
T1	6	1170.4	45.98	00.028
T2	6	3029.4	33.59	00.037
T3	6	1575.8	62.51	00.538
T4	6	2176.9	33.89	00.030
T5	6	2013.3	17.81	00.053
T6	6	1947.3	82.12	00.081

Table 7 TNF-α Difference Among Groups Compared to Control

Group	No. of Samples	Mean (pg/mL)	Std. Deviation	P value
C	6	5917	2.26
T1	6	896, 30	169.45	0.000
T2	6	247.03	17.33	0.006
T3	6	749.84	31.85	0.002
T4	6	16.4700	8.55	0.001
T5	6	530.67	53.51	0.000
T6	6	20.0117	2.49	0.0071

Figure 1 Immunohistochemical analysis of Cryptosporidium antigen in intestinal mice tissues of different groups: the control group and T1, T2, T3, T4, T5, and T6 were respectively represented in (A–G).

Figure 2 Immunohistochemical analysis of CD3 expression in intestinal mice tissues of different groups: the control group and T1, T2, T3, T4, T5, and T6 were represented in (A–G), respectively.

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