The Effect Of Intestinal Microflora Community Diversity In Preterm Infants With Feeding Intolerance

PartⅠ: The Effect of Intestinal Microflora Community Diversityin Preterm Infants with Feeding Intolerance by PCR-DGGE【Objective】Using the16s rDNA PCR denaturing gradient gelelectrophoresis（PCR-DGGE）,to investigate the alterations of intestinalmicroflora community diversity in premature infants with feedingintolerance(FI) and the relationship with the outcome of the disease.【Methods】Took20FI premature infants (from April2012to April2013)in Department of Neonatology of Children’s Hospital, ChongqingMedical University as FI group, and divided it into three subgroupsaccording to the definition of FI,5GRV of more than50%of the previousfeeding volume(FIⅠ),8emesis or abdominal distention or both(FIⅡ) and13the disruption of the patient’s feeding plan(FIⅢ). We took20FIpremature infants’ stool specimens within24hours of the symptom onset ofFI (t0) and after the recovery of FI (t1), and assessed bacterial diversity instool specimens within24hours of the symptom onset of FI (t0) and after the recovery of FI (t1) by16s rDNA PCR-DGGE. At the same time, wecollected the stool specimens of20feeding tolerance premature infants withthe same gestational age, birth weight and day-old as control（CⅠ、CⅡ andCⅢ）. Analysis the bands (S) and Shannon index(H′) of PCR-DGGEpicture by using the software of quantity one.【Results】①In contrast to the control group, the bands(S) ofintestinal bacterial was relatively lower in the FI group ［group Ⅰ（9.80±4.92vs19.20±2.95，P＜0.05）, groupⅡ（9.37±3.46vs15.00±3.82，P＜0.05），groupⅢ（10.69±3.04vs15±3.27，P＜0.05）］.②In contrast to the control group, Shannon index(H′) of intestinal bacterialwas relatively lower in the FI group ［group Ⅰ（2.16±0.51vs2.90±0.16，P＜0.05）,groupⅡ（2.14±0.42vs2.65±0.27，P＜0.05），groupⅢ（2.27±0.35vs2.61±0.25，P＜0.05）］.③While,therewasnostatisticalsignificant difference in S and H′between the recovery of FI (t1) and thecontrol group (P＞0.05).【Conclusion】 There was a decrease in intestinal microfloracommunity diversity in premature infants with FI, which recovered with therehabilitation of this disease. PartⅡ: The Effect of the Bacterial Species in Preterm Infantswith Feeding Intolerance by T-A Cloning Kit and Sequencing【Objective】Toinvestigatethedistributionofbacteriaandcommomdominant bacteria in stool specimens in preterm infants with FeedingIntolerance by T-A Cloning Kit and sequencing.【Methods】Recycled all bands from DGGE,and the result from T-Acloning and sequencing were comparatively analysised on NCBI(http://blast.ncbi.nlm.nih.gov/Blast).【Results】The result of T-A cloning and sequencing from DGGEshow that the main bacterial types of FI and control group were no difference,there are9Bacterial Species and Uncultured bacterium, include Klebsiella,Escherichia, Enterobacter, Streptococcus and so on. And compared withthe control group, the proportion of Klebsiella (46.82%vs38.66%,P＜0.05)and Bacteroides were higher in the FI group,while the proportion ofBifidobacterium, Lactobacillus, Escherichia and Enterobacter were lowerin the FI group.【Conclusion】Theproportionof Klebsiellawashigherandprobioticswas lower in intestinal microflora, which might be correlated with the onsetof FI in Preterm Infants.