ABSTRACT
The most crucial and expensive fragment in the broiler chicken production industry is the feed. Because of the rising demand, finding a cheap and effective feed is an urgent necessity. Fermentation of broiler feed by probiotic fungal starters can enhance the nutrient's availability and digestibility while preventing pathogenic growth. In this study different Rhizopus spp. have been isolated from agricultural soils around Izmir, Turkey, and tested for their probiotic potential and fermentative capacity. The isolated Rhizopus strains first underwent microscopical fluorescent investigation to exclude endofungal bacterial presence, then, those without endofungal bacteria (totally 82) were tested for antimicrobial activity counter bacterial and fungal pathogens. The ones with wide-spectrum antimicrobial activity (totally 10) were tested for gastrointestinal tolerance and antioxidant ability. Upon phenotypic and genotypic identification, the 10 isolates were found to belong to Rhizopus oryzae species. While all 10 strains showed variable gastrointestinal tolerance and antioxidant activities, three of them (92/1, 236/2, and 284) had relatively high antioxidant activity. Upon fermentative capacity assay, compared to unfermented commercial feed, there was a general decrease in crude fiber content by 56% after fermentation by 92/1 isolate for 4 days and 236/2 isolate for 2 days. The highest increase in crude protein content (by 14.5%) occurred after a 4-day fermentation period by 236/2 isolate. The highest increase in metabolizable energy was 8.64%, by the 284 isolate after 2 days of fermentation. In conclusion, the three strains showed good probiotic properties and fermentative capacities hence can be beneficial for the poultry industry.
Subject(s)
Animal Feed , Crops, Agricultural/microbiology , Fermentation , Probiotics , Rhizopus oryzae/metabolism , Animals , Anti-Bacterial Agents , Antifungal Agents , Antioxidants/metabolism , Chickens , Genotype , Phenotype , Rhizopus oryzae/genetics , Rhizopus oryzae/isolation & purification , Soil , Soil MicrobiologyABSTRACT
Rhizopus species are opportunistic pathogens and cause infections which lead to deaths in individuals with the weakened immune system. Some strains of Rhizopus species have been detected to have a symbiotic relationship with bacteria. The toxicity of the Rhizopus species is important. Because strains harbouring endofungal bacteria are able to produce secondary metabolites and if endofungal bacteria are released from mycelium, serious problems can occur. We aimed to investigate the presence of endofungal bacteria in Rhizopus species isolated from food samples. Rhizopus species were isolated from different food samples. The presence of endofungal bacteria in the Rhizopus isolates was investigated. Rhizopus strains containing the endofungal bacteria were identified through phenotypic and genotypic methods. Universal primers amplifying bacterial 16S rRNA region were used to amplify 1.2-1.5-kb fragment from fungal metagenomic DNA. Sequence analysis of PCR products amplified from fungal metagenomic DNA was made. Fluorescence microscopy and scanning electron microscopy were used to visualize the presence of endofungal bacteria in fungal hyphae. According to our results, the Rhizopus strains is associated with Serratia marcescens, Pseudomonas fluorescens and Klebsiella pneumoniae. Until now there is no evidence that Pseudomonas fluorescens and Klebsiella pneumoniae were identified as endofungal. These species are opportunistic pathogen dangerous for humans. It is important for humans not only the presence of the fungi but also the presence of the endofungal bacteria in foods. Our work is important because it draws attention to the presence of endofungal bacteria in foods. Because there is danger releasing of a bacterium from the mycelium, it is likely to face sepsis or serious problems.
