**2.6. Statistical analysis**

The results were presented as mean ± SE of three replicates. Prior to statistical analysis, all data were tested for normality of distribution using the Kolmogorov–Smirnov test. The homogeneity of variances among different treatments was tested using Bartlett's test. Then, data were subjected to one-way ANOVA to evaluate effects of *Lactobacillus acidophilus* supplementation. Differences between means were tested at the 5% probability level using Duncan test. The optimum *Lactobacillus acidophilus* level was determined using polynomial regression analysis. All the statistical analyses were done using SPSS program version 20 (SPSS, Richmond, VA, USA).

> *Pseudomonasa aeruginosa and Aeromonas hydrophila* were reduced significantly (p < 0.05) while *Bacillus* species was greatly increased. However, there was no significantly difference in the load of *Staphylococcus* species. The highest *Pseudomonas aeruginosa, Aeromonas hydrophila* and *Staphylococcus* species were recorded in prawn fed control diet while the least was observed in prawn fed diet fortified with 10<sup>5</sup> cfu/mL *Lactobacilus acidophilus*. However, the highest *Bacillus* species was obtained in prawn fed diet supplemented with 105 cfu/mL *Lactobacilus*

> Means (Log10 cgu/mL) with different superscripts are significantly different (p < 0.05), while, absence of letters means no

*Bacillus* species 4.98 ± 0.07a 6.08 ± 0.58 6.14 ± 0.02 6.15 ± 0.03 6.17 ± 0.41 6.26 ± 0.21

**Table 3.** *Gut microbiota composition of Macrobranchium vollenhovenii fed diets fortified with lactobacillus acidophilus*.

*Lactobacillus acidophilus* **inclusion levels (cfu/mL) Parameters (log10cfu/mL)**

significantly different (P > 0.05). TVC = Total viable bacteria count; TEB = Total enterobacterceae.

*Lactobacillus acidophilus* **inclusion levels (cfu/mL)**

**Table 2.** Gut microbiota of *Macrobranchium vollenhovenii* fed diets fortified with *Lactobacillus acidophilus*.

Control 6.32 ± 0.03a 6.12 ± 0.02 6.21 ± 0.04 5.96 ± 0.11 6.18 ± 0.20 5.72 ± 0.38 6.01 ± 0.16 5.62 ± 0.12 5.91 ± 0.09 5.43 ± 0.13a 5.81 ± 0.45 5.40 ± 0.90a

Means (Log10 cgu/mL) with different superscripts are significantly different (p < 0.05), while, absence of letters means no

**Control 101 102 103 104 105**

6.02 ± 0.09 5.92 ± 0.06 5.63 ± 0.26 5.57 ± 0.28a 5.32 ± 0.04a 5.15 ± 0.32a

6.06 ± 0.29a 5.49 ± 0.03a 5.38 ± 0.07 5.20 ± 0.22 4.90 ± 0.19 4.85 ± 0.04

6.24 ± 0.03 6.22 ± 0.91 6.20 ± 0.15 6.19 ± 0.33 6.18 ± 0.21 6.18 ± 0.05

**TVC\* TEB\***

http://dx.doi.org/10.5772/intechopen.78010

99

Gut Microbiota and Innate Immune Response of *Macrobrachium vollenhovenii* Infected…

Effects of *Lactobacillus acidophilus* based diets on innate immune responses of *Macrobranchium vollenhovenii* was presented in **Table 4**. The immune response of the prawn was stimulated by *Lactobacillus acidophilus* supplementation. The SOD, CAT, RBA, THC, PO, ROI and lysozyme activity were significantly different (p < 0.05) among the treatments. Highest SOD, CAT, RBA, THC, PO, ROI and lysozyme activity were recorded in prawn fed 10<sup>5</sup> cfu/mL diet; meanwhile,

the lowest values were obtained in the control group.

*acidophilus.*

**Parameters (log10cfu/mL)**

*Pseudonomas aeruginosa*

*Aeromonas hydrophila*

*Staphylococcus* species

significantly different (P > 0.05).
