| Noise is a potential but not negligible environmental factor in animal husbandry.Noise has been described as unwanted sound.And chickens have complex cognition,whether the environment sound in livestock farm is noise depends on the subjective concept of chickens,and the environment sound in livestock farm effects on livestock and poultry needs further study.In order to explore the effects of the environment sound in livestock farm on intestinal barrier function and intestinal flora colonization of pullets,1-day-old Roman white pullets were selected as the research object.336 pullets were divided into three groups: control group(no additional sound added),low intensity sound group(65-75dB,LIS),and high intensity sound group(85-95dB,HIS).They were raised in three sound proof chambers.The treatment group was subjected to sound stimulation at the age of1 day,which recorded by the local laying farm,6 hours per day,1 hour interval,1 hour sound stimulation,and the sound stimulation lasted for 4 weeks.The duodenum,jejunum,ileum tissues and cecum contents of birds were collected every week.The morphological and structural changes of ileum tissues were observed by HE staining,the expression levels of claudin-1,Occludin,ZO-1and Mucin2 in ileum tissues were detected by immunofluorescence.Detection of tight junction proteins and Mucin2,heat shock proteins,immunoglobulin,cytokines,Toll-like receptors m RNA expression levels in intestinal tissues of duodenum,jejunum and ileum by Quantitative Real-time PCR(q RT-PCR).The structure of intestinal microflora in cecum was analyzed by 1 16 SrDNA sequencing.After 1 week of the envirounment sound in livestock farm stimulation,in ileum,LIS increased the m RNA expression levels of claudin-1,E-cadherin,Occludin,Mucin2 and ZO-2(P<0.05),HIS increased the m RNA expression levels of claudin-1,claudin-4,E-cadherin and ZO-1 in duodenum(P<0.05).After 2 weeks of sound stimulation,in ileum HIS increased the m RNA expression levels of Claudin-4,Occludin,ZO-1 and ZO-2(P<0.05).After 3 weeks of sound stimulation,the expression level of Occludin and in duodenum of LIS group increased significantly(P<0.05),the m RNA expression levels of E-cadherin,Mucin2,ZO-1 and ZO-2 showed an upward trend(P>0.05),LIS and HIS decreased the m RNA expression levels of claudin-1,claudin-4,E-cadherin,Occludin,Mucin2,ZO-1 and ZO-2 in ileum(P<0.05),there was no significant difference between LIS and HIS groups(P>0.05).After 4 weeks of sound stimulation,m RNA expression levels of claudin-4in duodenum and tight junction protein and mucin in ileum were decreased in the LIS and HIS groups(P<0.05),there was no significant difference between the two groups(P<0.05).Histopathological sections of ileum showed that inflammatory cell infiltration in ileum of LIS and HIS groups decreased after 1 week of sound stimulation,and the number and height of intestinal villi increased after 2 weeks of sound stimulation.Immunofluorescence results showed that sound stimulation increased the protein expression of Occludin,Mucin2 and ZO-1 after 1 week(P<0.05).In LIS group the protein expression of Occludin,Mucin2 and ZO-1 increased after 2 weeks(P <0.05),the protein expression of Occludin,Mucin2,ZO-1 decreased after 3 weeks of sound stimulation(P<0.05),and the protein expression of Claudin-1,Occludin,MUCIN2,ZO-1decreased after 4 weeks of sound stimulation(P< 0.05),which of HIS group was significantly lower than those of LIS group(P<0.05).The m RNA expression level of HSP40,HSP60 and HSP90 in duodenum and HSP40 and HSP60 in ileum had no significant change after 1 week of LIS stimulation(P>0.05),the m RNA level of HSP70 in duodenum and HSP70 and HSP90 in ileum were significantly decreased(P<0.05).The m RNA expression level of HSPs in duodenum and ileum was significantly decreased after 1 week of HIS,and was lower than that in LIS group(P< 0.05).There was no significant diffrences in the expression of HSPs in duodenum and ileum after 2 weeks of sound(P>0.05).After 3 weeks of sound stimulation,LIS and HIS decreased the m RNA expression of HSP90 in the duodenum(P <0.05),HIS increased the m RNA expression levels of HSP40 and HSP60 in ileum(P<0.05),but the expression levels of HSP70 and HSP90 were not significantly changed(P>0.05).After 4 weeks of sound stimulation,the m RNA expression levels of HSP70 and HSP90 in duodenum and HSPs in ileum were decreased by sound stimulation(P<0.05),the m RNA expression levels of HSP70 and HSP90 in ileum of HIS group were lower than those of LN group(P<0.05).The m RNA expression levels of NF-κB,PTGES,COX-2,iNOS,IL-1β,IFN-γ and TNF-α in ileum were significantly increased after 1 week of low decibel sound stimulation(P<0.05);After 1week of high decibel sound stimulation,the m RNA expression levels of TLR4,NF-κB,PTGES,COX-2,iNOS in duodenum and NF-κB,PTGES,COX-2,iNOS,IL-1β and TNF-α in ileum were significantly increased(P<0.05).The m RNA expression levels of NF-κB,PTges,COX-2,iNOS,IFN-γ and TNF-α in ileum were significantly increased after 2 weeks of high decibel sound stimulation(P<0.05).The m RNA expression levels of TLR4,NF-κB,PTGES,iNOS,IL-1β and IFN-γ in ileum were significantly decreased after 3 weeks of low decibel sound stimulation(P <0.05);The m RNA expression levels of NF-κB,PTGES and iNOS in duodenum and TLR4,NF-κB,PTGES,iNOS and IL-1β in ileum were significantly decreased after 3 weeks of high decibel sound stimulation(P<0.05).The expression levels of NF-κB,PTGES,COX-2,iNOS,IL-1β,TNF-α in duodenum and TLR4,NF-κB,PTGES,COX-2,iNOS,IL-1β,IFN-γ,TNF-α in ileum were significantly decreased after 4 weeks of high and low decibel sound stimulation(P<0.05).However,the m RNA expression levels of tight junction proteins,mucins,HSPs and inflammation-related factors in jejunum were not significantly affected by sound stimulation(P <0.05),suggesting that sound stimulation had tissue-specific effects on the intestinal tract of pullets.α-diversity analysis and β-diversity analysis showed that sound had no significant effect on the richness and diversity of intestinal microflora of pullets.There were no significant differences in Chao1 index and Shannon index after1 week,2,3 and 4 weeks of sound stimulation.PCo A results showed that there was no significant separation between the three groups.Species annotation information showed that sound stimulation for 1 week and 2 weeks decreased the abundance of Actinomyces at the phylum level(P<0.05),the abundance of Actinomycota increased after 3 and4 weeks of sound stimulation(P<0.05).Sound exposure increased Enterococcus,Subdoligranulum,and Laospirillum(P<0.05).In conclusion,Sound from of the farm can promote the development of intestinal barrier and the colonization of beneficial bacteria in pullets in short order.The effects of sound with different loudness(65-75dB and 85-95dB)have no significant diffrents.The effects of sound on intestinal tissue are specific,and jejunum is less affected by sound. |
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