**3.4 Isolation and characterization strategies of probiotics for mass-reared** *Ceratitis capitata*

The majority of probiotics have thus far been isolated from medfly using the classical methods. Culture-dependent approaches have been used and adjusted to isolate and identify most of the probiotics. In the culture-dependent approach, the culture is using solid media allowing growth of bacteria such as Luria Bertani (LB), tryptic soy agar (TSA) [28], or a chromogenic medium such as CHROMagar orientation [16]. However, the morphological characterization by itself is unresponsive because bacteria's morphological characteristics, such as their color and shape, are not always constant. Further accurate identification approaches have been used such as the 16SrRNA gene amplification and sequencing. To reassemble bacterial colonies in haplotypes while minimizing sequencing, Hamden et al., [35] used the universal primers S-D-Bact-1494-a-20 and L-D-Bact-0035-a-15 to perform DNA amplification of the 16S–23S rRNA internal transcribed spacers region (ITS-PCR) (**Table 3**). While Augustinos et al., [29] combined morphological examination of colonies and RFLP assays, Ben Ami et al., [16] chose amplified rDNA restriction analysis (ARDRA), both techniques are based on restriction enzymes that provide the same digestion pattern.


### **Table 3.**

*Isolation and selection approaches of probiotics for medfly mass-rearing.*

*Probiotics as a Beneficial Modulator of Gut Microbiota and Environmental Stress… DOI: http://dx.doi.org/10.5772/intechopen.110126*
