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Salmonella Enterica Serovars thaum Tsis Muaj TtrA Thiab PduA Genes Txhim Kho Cov Tshuaj Tiv Thaiv Kab Mob Thaum Ntxov Kab Mob

Oct 24, 2023

Salmonella spp. yog ib qho ntawm cov kab mob loj tshaj plaws uas ua rau muaj kev puas tsuaj rau kev lag luam nqaij qaib thiab ua rau muaj kev cuam tshuam rau pej xeem kev noj qab haus huv ib yam nkaus. Ob qho tib si muaj peev xwm ciaj sia nyob hauv txoj hnyuv ib puag ncig thaum mob nrog rau lawv txoj kev sib raug zoo nrog lub cev tiv thaiv kab mob, ua lub luag haujlwm tseem ceeb thaum kis kab mob hauv cov nqaij qaib. Lub hom phiaj ntawm txoj kev tshawb no yog txhawm rau txheeb xyuas qhov muaj nyob ntawm macrophages thiab CD4+ / CD8+ cov hlwb siv cov txheej txheem immunohistochemistry, hauv cov kab lag luam ntawm cov qaib tau sim kis los ntawm cov tsiaj qus thiab mutant hom ntawm Salmonella. Enteritidis thiab Salmonella Typhimurium tsis muaj ttrA thiab pduA noob. Salmonella Enteritidis ∆ttrA∆pduA ua rau muaj feem pua ​​​​ntawm thaj chaw stained ntau dua li hom tsiaj qus, tshwj tsis yog cov hens teeb. Salmonella Typhimurium wild-type strain thiab Salmonella Typhimurium ∆ttrA∆pduA cov kab mob ua rau muaj cov qauv zoo sib xws uas, ntawm 1 thiab 14 dpi, caecal tonsils thiab ileum ntawm cov noog tau pom ntau qhov chaw stained piv rau 3 thiab 7 dpi. Nyob rau hauv tag nrho cov kab mob uas tau kawm, qhov tseem ceeb infiltration ntawm macrophages nyob rau hauv kev sib piv nrog CD{10}} thiab CD{11}} hlwb tau pom. Zuag qhia tag nrho, cov tsiaj kis los ntawm cov kab mob mutant pom qhov zoo stained cheeb tsam siab dua li hom tsiaj qus. Kev tshem tawm hauv ob qho tib si ttrA thiab pduA noob tau ua rau muaj kev cuam tshuam ntau dua ntawm macrophages thiab CD4+ thiab CD8+ hlwb hauv cov noog host, qhia tias tsis muaj kab mob attenuation, txawm nyob hauv ntau hom Salmonella.

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cistanche cov txiaj ntsig rau cov txiv neej-ua kom muaj zog tiv thaiv kab mob

Salmonella enterica yog cov khoom noj uas ua rau muaj kev puas tsuaj rau tsiaj nyeg thiab cuam tshuam ncaj qha rau pej xeem noj qab haus huv. Salmonella enterica subsp. enterica serovar Enteritidis (Salmonella Enteritidis) thiab Typhimurium (Salmonella Typhimurium) tau cuam tshuam nrog cov khoom noj khoom haus ntau xyoo lawm. Nyob nruab nrab ntawm 1995 thiab 2010, Salmonella Enteritidis tau txheeb xyuas hauv 34,2% ntawm tag nrho cov qauv zoo rau Salmonella spp1. Tsis tas li ntawd, Lub Caij Ntuj Sov thiab al.2 tau luam tawm ib qho kev tshawb fawb tseem ceeb tshawb xyuas lub luag haujlwm ntawm tetrathionate-encoding gene, xaiv cov serotype Salmonella Typhimurium tab sis siv cov nas ua qauv sim. Ua raws li qhov no rau hauv tus account, kev nkag siab txog peb txoj kev paub ntawm tus tswv tsev-cov kab mob sib cuam tshuam tuaj yeem pab txhim kho kev tswj hwm thiab kev ntsuas kev tshem tawm. Ntau yam muaj feem cuam tshuam nrog salmonellosis pathogenesis, xws li lub peev xwm ntawm cov kab mob kom rov ua dua nyob rau hauv ib qho chaw mob mucous, nyob ntawm kev tau txais cov khoom noj thiab kev ua pa anaerobic2. Txawm li cas los xij, kev muaj cov khoom noj tsis tau lees tias cov kab mob muaj sia nyob hauv ib puag ncig kev sib tw nrog cov neeg nyob ze ntawm lwm cov kab mob. Yog li, lub peev xwm ntawm Salmonella enterica rau metabolize tetrathionate siv tetrathionate reductase los tsim 1,2-propanediol raws li lub zog qhov chaw muaj txiaj ntsig zoo. Cov enzyme no yog tsim los ntawm TtrA, TtrB, thiab TtrC. Thawj subunit raug suav hais tias yog nyob rau hauv molybdopterin (MPT) superfamily thiab muaj FeS bounding domain uas koom nrog hauv kev txo cov tetrathionate rau hauv thiosulfate (S2O3 2−)2–4. Te 1,2-propanediol yog siv los ntawm cov kab mob microcompartments (MCP). Cov qauv no yog tsim los ntawm xya cov protein sib txawv, ntawm cov PduA yog qhov tseem ceeb ntawm MCP qauv5. Ua ntej, 1, 2-propanediol hloov mus rau propionaldehyde, uas nyob rau hauv lem yog txo mus rau propanol thiab propionate los ntawm propanediol dehydratase kev ua. Cov txheej txheem no tsim ATP los ntawm phosphorylation, ib qho hluav taws xob (1-}propanol) gradient rau NAD regeneration, thiab tus neeg nruab nrab (propionyl-CoA) uas tuaj yeem siv los ua cov pa roj carbon thiab lub zog thoob plaws methyl citrate los ntawm, nyob ntawm B12 vitamin synthesized nyob rau hauv ib tug endogenous yam 6. Thaum kis kab mob nqaij qaib, lub peev xwm ntawm Salmonella enterica serovars tuaj yeem cuam tshuam thiab ciaj sia nyob rau hauv cov hnyuv epithelial hlwb thiab macrophages yog ua raws li kev tiv thaiv kab mob evasion7. Nyob rau hauv cov ntsiab lus no, tus kab mob yog ib qho tseem ceeb theem uas nyob ntawm kev sib cuam tshuam ntawm cov kab mob thiab cov tswv tsev cell thiab cov kab mob muaj peev xwm kov yeej lub plab hnyuv epithelium teeb meem los lav nws colonization thiab replication. Txawm li cas los xij, nws ua rau cov kab mob inflammatory thiab tiv thaiv kab mob 8 uas ua rau endocytosis thiab phagocytosis los ntawm epithelial thiab antigen-presenting cells (APCs), feem. Cov tshuaj tiv thaiv kab mob ntawm cov hlwb no ua rau muaj qhov tshwm sim hauv lub cev los ntawm macrophages thiab kev hloov kho lub cev tiv thaiv kab mob sib sau ua ke ntawm CD4+ thiab CD8+ ua kom 9. Txhawm rau tso lub teeb ci ntawm tus tswv tsev-cov kab mob sib cuam tshuam tom qab kab mob plab hnyuv los ntawm Salmonella hauv qaib, peb ntsuas cov pej xeem ntawm lub cev tiv thaiv kab mob thaum lub sij hawm plab hnyuv thiab kab mob hauv cov noog ntawm cov kab lag luam tawm tsam los ntawm cov tsiaj qus mutant hom ntawm Salmonella Enteritidis thiab Salmonella Typhimurium. , nqa deletions hauv cov noob muaj feem xyuam rau cov metabolization ntawm tetrathionate (ttrA) thiab 1, 2-propanediol (PDU).

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Cov txiaj ntsig

Kev sim 1-Salmonella Enteritidis kev sib tw.

Cov txiaj ntsig ntawm kev txheeb xyuas qhov muaj lub cev tiv thaiv kab mob hauv cov kab mob caecal tonsils, caecum, ileum, thiab daim siab los ntawm broiler, lub teeb-ntev hens, thiab semi-heavy laying hens muaj nyob rau hauv Tables 1, 2, thiab 3. Peb pom ntau CD 4+ thiab macrophage cell infiltration los ntawm broilers sib tw nrog Salmonella Enteritidis ∆ttrA∆pduA (SEΔttrAΔpduA) dua li ntawm broilers sib tw nrog Salmonella Enteritidis qus-hom hom (wt-SE) los yog cov noog tsis muaj kab mob, nyob rau hauv tag nrho cov ntaub so ntswg soj ntsuam. Ib qho kev zam tau pom rau CD4+ cell infiltration ntawm 3 dpi hauv caecal tonsils, ileum, thiab siab, thiab rau macrophage infiltration ntawm 3 thiab 14 dpi hauv lub siab los ntawm SEΔttrAΔpduA sib tw broilers. Ntxiv mus, tus naj npawb ntawm CD8+ hlwb infiltrated tau pom nyob rau hauv ntau dua los ntawm cov noog sib tw nrog SEΔttrAΔpduA ntawm 1 thiab 7 dpi hauv caecum thiab siab, ntawm 3 dpi hauv caecal tonsils, thiab 14 dpi hauv ileum. Nyob rau hauv sib piv, wt-SE-kev sib tw noog muaj siab infiltration ntawm CD{20}} hlwb nyob rau hauv lub ileum thiab caecal tonsils ntawm 1 thiab 7 dpi, raws li (Tab. 1; Supplementary Fig. S1). Table 2, thiab Ntxiv Fig. S2 qhia cov CD4+ thiab CD8+, thiab macrophage infiltration pom nyob rau hauv cov ntaub so ntswg ntawm ib nrab hnyav nteg qe. Feem ntau, muaj kev hloov pauv loj ntawm thaj chaw ntawm cov cell infiltrates txog ob qho kev sib tw thiab cov ntaub so ntswg kawm. Cov noog sib tw nrog SEΔttrAΔpduA pom cov cheeb tsam siab dua los ntawm CD4+ hlwb (hauv ileum ntawm 1 thiab 14 dpi, thiab siab ntawm 7 dpi), thiab macrophages (hauv ileum ntawm 7 dpi, thiab siab ntawm 3, 7, thiab 14 dpi), dua li nyob rau hauv tib cov ntaub so ntswg ntawm wt-SE-kev sib tw noog. Ntawm qhov tod tes, CD8+ hlwb tau pom nyob rau hauv ntau ntau hauv caecal tonsils (ntawm 1 thiab 7 dpi) thiab caecum (ntawm 3 dpi) ntawm cov noog sib tw nrog wt-SE hom. Kev sib txawv ntawm cov qaib broiler thiab ib nrab hnyav nteg qe qaib, Salmonella Enteritidis ∆ttrA∆pduA ua rau lub cev tsis muaj zog tiv thaiv kab mob tsawg dua li cov tsiaj qus, hauv kev sib tw los ntawm lub teeb nteg hens. Wt-SEchallenged noog pom loj CD4+ infiltration thaj chaw hauv caecal tonsils (ntawm 1 thiab 3 dpi), caecum (ntawm 3, 7, thiab 14 dpi), ileum (ntawm 1 thiab 7 dpi), thiab siab (ntawm 14 dpi). Ib yam li ntawd, cov kev sib tw nrog SEΔttrAΔpduA tau ua rau txo qis qhov chaw nkag ntawm ob lub CD8+ hauv caecal tonsils (ntawm 1 dpi) thiab ileum (ntawm 7 thiab 14 dpi), thiab macrophages hauv caecal tonsils thiab caecum (ntawm 3 dpi), nyob rau hauv kev sib piv nrog wt-SE-chaw noog. Tsis muaj kev hloov pauv tseem ceeb ntawm lub cev tiv thaiv kab mob hauv cheeb tsam ntawm CD8+ thiab cov hlwb macrophage tau pom hauv daim siab los ntawm ob qho tib si SEΔttrAΔpduA thiab wt-SE-kev sib tw noog. Cov ncauj lus kom ntxaws ntawm qhov kev sib tw nrog lub teeb-ntev hens muaj nyob rau hauv Table 3 (saib Ntxiv Fig. S3).

Table 1.

Table 1. Kev nthuav qhia ntawm qhov sib txawv ntawm qhov tseem ceeb muaj feem cuam tshuam txog kev faib tawm ntawm cov kab mob sib txawv ntawm cov kab mob sib txawv hauv cov kab mob ntawm cov kab mob broilers kis tus kab mob Salmonella Enteritidis qus thiab mutant hom ntawm ntau hnub tom qab kis kab mob. DPI, hnub tom qab kis kab mob; ∆-SE, Salmonella Enteritidis ∆ttrA∆pduA; wt-SE, Salmonella Enteritidis qus-hom; ns, tsis muaj qhov txawv txav. Hauv txhua lub cev thiab DPI, * txhais tau tias sib txawv los ntawm ob txoj kev ANOVA ua raws li Bonferroni qhov kev sib piv ntawm cov tsiaj qus thiab mutant hom (*P Tsawg dua lossis sib npaug rau 0.05; **P Less dua lossis sib npaug rau 0.01; ***P Tsawg dua lossis sib npaug rau 0.001; ****P Tsawg dua lossis sib npaug rau 0.0001). Te strain nthuav tawm nyob rau hauv lub rooj (∆-SE los yog wt-SE) raws li qhov tseem ceeb nyob rau hauv lub cev thiab DPI yog qhia qhov loj infltration cheeb tsam rau txhua lub cell.

Table 2.

Table 2. Kev nthuav qhia ntawm qhov sib txawv ntawm qhov tseem ceeb muaj feem cuam tshuam txog kev faib tawm ntawm cov kab mob sib txawv ntawm cov kab mob sib txawv hauv cov kab mob ntawm cov kab mob ib nrab hnyav uas kis tus kab mob Salmonella Enteritidis qus thiab mutant hom nyob rau ntau hnub tom qab kis kab mob. DPI, hnub tom qab kis kab mob; ∆-SE, Salmonella Enteritidis ∆ttrA∆pduA; wt-SE, Salmonella Enteritidis qus-hom; ns, tsis muaj qhov txawv txav. Hauv txhua lub cev thiab DPI, * txhais tau tias sib txawv los ntawm ob txoj kev ANOVA ua raws li Bonferroni qhov kev sib piv ntawm cov tsiaj qus thiab mutant hom (*P Tsawg dua lossis sib npaug rau 0.{{10}}5 ; **P Tsawg dua lossis sib npaug rau 0.01; ***P Tsawg dua lossis sib npaug rau 0.001; ****P Tsawg dua lossis sib npaug rau 0.0001). Te strain nthuav tawm nyob rau hauv lub rooj (∆-SE los yog wt-SE) raws li qhov tseem ceeb nyob rau hauv lub cev thiab DPI yog qhia qhov loj infltration cheeb tsam rau txhua lub cell.


Table 3.


Table 3. Kev sawv cev ntawm qhov sib txawv tseem ceeb uas cuam tshuam txog kev faib tawm ntawm cov kab mob sib txawv ntawm lub cev tiv thaiv kab mob hauv cov kab mob ntawm lub teeb nteg qe qaib uas kis tus kab mob Salmonella Enteritidis qus thiab mutant hom kab mob sib txawv ntawm cov hnub sib txawv tom qab kis kab mob. DPI, hnub tom qab kis kab mob; ∆-SE, Salmonella Enteritidis ∆ttrA∆pduA; wt-SE, Salmonella Enteritidis qus-hom; ns, tsis muaj qhov sib txawv tseem ceeb. Hauv txhua lub cev thiab DPI, * txhais tau hais tias qhov sib txawv ntawm ob txoj kev ANOVA ua raws Bonferroni qhov kev sib piv ntawm cov tsiaj qus thiab mutant hom (*P Tsawg dua lossis sib npaug rau 0.{{10}}5 ; **P Tsawg dua lossis sib npaug rau 0.01; ***P Tsawg dua lossis sib npaug rau 0.001; ****P Tsawg dua lossis sib npaug rau 0.0001). Cov kab mob uas tau nthuav tawm hauv lub rooj (∆-SE lossis wt-SE) yog qhov tseem ceeb hauv lub cev thiab DPI yog qhov qhia tau hais tias qhov chaw nkag mus rau txhua lub cell.

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cistanche cog-nce kev tiv thaiv kab mob

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Kev sim 2-Salmonella Typhimurium kev sib tw.

Cov txiaj ntsig ntawm kev txheeb xyuas qhov muaj lub cev tiv thaiv kab mob hauv cecal tonsils, caecum, ileum, thiab daim siab los ntawm broilers, semi-heavy nteg hens, thiab lub teeb nteg hens tau pom nyob rau hauv Tables 4, 5, thiab 6, ntsig txog. Zuag qhia tag nrho, broilers kis los ntawm cov kab mob sib kis tau pom qhov chaw cim zoo siab dua li cov neeg sib tw nrog Salmonella Typhimurium wild-type strain (wt-STM) strain, ntawm tag nrho plaub hnub kuaj. Tsis tas li ntawd, tsis muaj kev hloov pauv ntawm lub cev tiv thaiv kab mob hauv cov pab pawg tswj tsis muaj kab mob. Te CD4+ cov cheeb tsam ntawm tes hauv txhua cov ntaub so ntswg ntawm broilers tau ncav cuag qhov sib txawv ntawm qhov sib txawv ntawm 1 dpi, nrog cov infiltrations loj dua rau kev sib tw nrog Salmonella Typhimurium ∆ttrA∆pduA (STM∆ttrA∆pduA). Ib qho tseem ceeb sib txawv ntawm thaj tsam ntawm lub cev tiv thaiv kab mob ntau npaum li cas los ntawm STM∆ttrA∆pduA- thiab wt-STM-cov noog sib tw tau, nrog cov kab mob loj dua thaum muaj kev sib tw nrog cov kab mob mutant hauv caecum thiab ileum ntawm 1 dpi, thiab daim siab. ntawm 14 dpi (CD{13}} hlwb); nyob rau hauv caecum, ileum, thiab daim siab ntawm 1 thiab 14 dpi (macrophages) (Table 4; Supplementary Fig. S4). Te tau los ntawm ib nrab hnyav nteg noog tsis pom qhov sib txawv ntawm mutant thiab qus-hom kev sib tw rau CD{20}} hlwb hauv caecum, thiab CD8+ hlwb hauv caecal tonsils thiab ileum nyob rau hauv tag nrho 4 hnub. kev soj ntsuam kab mob tom qab (Table 5; Supplementary Fig. S5). Txawm li cas los xij, thaum pom qhov tseem ceeb ntawm thaj tsam ntawm lub cev tiv thaiv kab mob hauv lub cev tau pom cov hens ib nrab hnyav nteg tau sib tw los ntawm STM∆ttrA∆pduA: loj macrophages infiltration cheeb tsam hauv tag nrho cov ntaub so ntswg kawm (ntawm 1 thiab 14 dpi); ib cheeb tsam loj infiltration ntawm CD4+ hlwb nyob rau hauv caecal tonsils thiab siab (ntawm 1, 7, thiab 14 dpi) (Table 5; Supplementary Fig. S5). Table 6, thiab ntxiv Fig. S6 qhia cov CD4+, CD8+, thiab macrophage infiltration pom nyob rau hauv cov ntaub so ntswg ntawm lub teeb nteg hens. Cov noog sib tw nrog STM∆ttrA∆pduA pom cov kab mob hauv lub cev loj hauv cheeb tsam CD4+ thiab macrophage hlwb hauv caecal tonsils thiab caecum (ntawm 14 dpi), thiab ileum (ntawm 1 thiab 14 dpi) piv rau lub cev tiv thaiv kab mob. thaj tsam ntawm tib cov ntaub so ntswg los ntawm wt-STM-chaw noog. Tsis muaj qhov sib txawv ntawm CD8+ thaj chaw infiltration tau pom nyob rau hauv caecal tonsils, caecum, thiab ileum los ntawm cov noog sib tw. Hauv qhov sib piv rau cov txiaj ntsig tau txais hauv lwm cov ntaub so ntswg, lub siab los ntawm STM∆ttrA∆pduA-cov noog sib tw muaj ntau qhov chaw ntawm lub cev tiv thaiv kab mob rau CD4+ thiab CD8+ hlwb ntawm tag nrho plaub dpi, thiab macrophages ntawm 1 thiab 14 dpi.

Table 4

Table 4. Kev nthuav qhia ntawm qhov sib txawv tseem ceeb ntsig txog kev faib tawm ntau ntawm cov kab mob tiv thaiv kab mob sib txawv hauv cov kab mob broilers kis tus kab mob Salmonella Typhimurium qus thiab mutant hom nyob rau ntau hnub tom qab kis kab mob. DPI, hnub tom qab kis kab mob; ∆-STM, Salmonella Typhimurium ∆ttrA∆pduA; wt-STM, Salmonella Typhimurium qus-hom; ns, tsis muaj qhov txawv txav. Hauv txhua lub cev thiab DPI, * txhais tau tias sib txawv los ntawm ob txoj kev ANOVA ua raws li Bonferroni qhov kev sib piv ntawm cov tsiaj qus thiab mutant hom (*P Tsawg dua lossis sib npaug rau 0.05; **P Less dua lossis sib npaug rau 0.01; ***P Tsawg dua lossis sib npaug rau 0.001; ****P Tsawg dua lossis sib npaug rau 0.0001). Te strain nthuav tawm nyob rau hauv lub rooj (∆-STM los yog wt-STM) raws li qhov tseem ceeb hauv lub cev thiab DPI yog qhov qhia qhov loj infltration cheeb tsam rau txhua lub cell.

Table 5

Table 5. Kev nthuav qhia ntawm qhov sib txawv tseem ceeb uas cuam tshuam txog kev faib tawm ntau ntawm cov kab mob sib txawv ntawm cov kab mob hauv lub cev ntawm ib nrab hnyav nteg qes kis tus kab mob Salmonella Typhimurium qus thiab mutant hom ntawm txawv hnub tom qab kis kab mob. DPI, hnub tom qab kis kab mob; ∆-STM, Salmonella Typhimurium ∆ttrA∆pduA; wt-STM, Salmonella Typhimurium qus-hom; ns, tsis muaj qhov txawv txav. Hauv txhua lub cev thiab DPI, * txhais tau tias sib txawv los ntawm ob txoj kev ANOVA ua raws li Bonferroni qhov kev sib piv ntawm cov tsiaj qus thiab mutant hom (*P Tsawg dua lossis sib npaug rau 0.{{10}}5 ; **P Tsawg dua lossis sib npaug rau 0.01; ***P Tsawg dua lossis sib npaug rau 0.001; ****P Tsawg dua lossis sib npaug rau 0.0001). Te strain nthuav tawm nyob rau hauv lub rooj (∆-STM los yog wt-STM) raws li qhov tseem ceeb hauv lub cev thiab DPI yog qhov qhia qhov loj infltration cheeb tsam rau txhua lub cell.

Table 6


Table 6. Kev sawv cev ntawm qhov sib txawv tseem ceeb uas cuam tshuam txog kev faib tawm ntawm cov kab mob sib txawv ntawm lub cev tiv thaiv kab mob hauv cov kab mob ntawm cov hens teeb pom kev zoo uas kis tus kab mob Salmonella Typhimurium qus thiab mutant strains ntawm ntau hnub tom qab kis kab mob. DPI, hnub tom qab kis kab mob; ∆-STM, Salmonella Typhimurium ∆ttrA∆pduA; wt-STM, Salmonella Typhimurium qus-hom; ns, tsis muaj qhov sib txawv tseem ceeb. Hauv txhua lub cev thiab DPI, * txhais tau hais tias qhov sib txawv ntawm ob txoj kev ANOVA ua raws Bonferroni qhov kev sib piv ntawm cov tsiaj qus thiab mutant hom (*P Tsawg dua lossis sib npaug rau 0.{{10}}5 ; **P Tsawg dua lossis sib npaug rau 0.01; ***P Tsawg dua lossis sib npaug rau 0.001; ****P Tsawg dua lossis sib npaug rau 0.0001). Cov kab mob uas tau nthuav tawm hauv lub rooj (∆-STM lossis wt-STM) yog qhov tseem ceeb hauv lub cev thiab DPI uas qhia tau hais tias qhov loj infiltration cheeb tsam rau txhua lub cell.

Kev sib tham

Cov kab mob, thaum raug anaerobic tej yam kev mob, yuav siv tau tetrathionate thiab 1,2-propanediol metabolic substrates rau lub zog thiab ua pa qhov chaw10. Yog li, Salmonella spp. tau ntev tau raug tshawb xyuas txog yuav ua li cas tshem tawm cov noob uas paub tias yog lub luag haujlwm rau txoj hauv kev no yuav cuam tshuam rau lawv txoj sia nyob hauv tus tswv tsev. Rau qhov zoo tshaj plaws ntawm peb txoj kev paub, tsuas yog ib qho kev tshawb fawb tshawb xyuas tib lub sijhawm tetrathionate- thiab propanediol-encoding gene cwj pwm tau luam tawm. Peb pab pawg tshawb fawb tau tshaj tawm txog qhov cuam tshuam ntawm cov kev tshem tawm no los ntawm kev soj ntsuam cov kab mob hauv lub cev thiab fecal excretion ntawm Salmonella Enteritidis thiab Salmonella Typhimurium hauv cov kab lag luam ntawm cov menyuam qaib11. Txhawm rau kom muaj kev sib tham txog cov ntsiab lus no, cov txiaj ntsig tam sim no tau qhia txog lub cev tiv thaiv kab mob nkag mus rau hauv cov ntaub so ntswg sib txawv ntawm cov qaib cov kab mob sib tw nrog ob hom tsiaj qus thiab cov kab mob mutant nqa deletions hauv ttrA thiab pduA noob. Tshaj li ntawm 2-lub lim tiam kev sim, qhov zoo stained thaj chaw ntawm CD4+ thiab CD8+ hlwb thiab macrophages ua raws li feem ntau cov qauv zoo sib xws, qhov twg ntawm 1 thiab 14 dpi nthuav tawm ntau dua ntawm kev tiv thaiv kab mob. hlwb. Qhov no tuaj yeem piav qhia los ntawm kev sib cuag ntawm tus tswv tsev tiv thaiv thaum cov kab mob nkag mus. Daim ntawv tshaj tawm dhau los tau qhia tias nyob rau hauv qaib cov kab mob, txawm tias thaum Salmonella tsis tawm ntawm 12 dpi, tus kab mob tuaj yeem ua tau zoo los ntawm cloacal swab los ntawm 13 dpi12, piav qhia vim li cas lub cev tiv thaiv kab mob hauv cheeb tsam ntawm 3 thiab 7 dpi tau qis dua tab sis rov qab mus rau qhov nce ntxiv. . Thaum xub thawj siab ib muag, nws yuav xav tias cov lus teb ntawm tus tswv tsev yuav raug txo qis thaum ob qho tib si pduA thiab ttrA raug tshem tawm vim cov noob no ua lub luag haujlwm tseem ceeb hauv kev ciaj sia thaum kis kab mob los ntawm Salmonella2,4,13-15. Txawm li cas los xij, peb cov txiaj ntsig tau muab piv rau ob tus mutant uas tsis muaj ob lub noob tau pom qhov sib txawv, cov kab mob sib kis ntawm Salmonella Enteritidis thiab Salmonella Typhimurium ua rau lub cev tiv thaiv kab mob ntau dua li cov tsiaj qus hom.

Cistanche deserticola—improve immunity (5)

Cov txiaj ntsig ntawm cistanche tubulosa- ua kom muaj zog tiv thaiv kab mob

Ib cheeb tsam luv tshaj plaws tuaj yeem ua rau muaj coob tus neeg nyob hauv txoj hnyuv, txheeb xyuas qhov kev tshawb fawb yav dhau los, nyob rau hauv Salmonella Enteritidis ∆ttrA∆pduA thiab Salmonella Typhimurium ∆ttrA∆pduA strains tau rov qab los ntawm cov lej ntau dua los ntawm cloacal swabs correlated 1 hom tsiaj qus. . Salmonella tuaj yeem coj tus cwj pwm zoo li cov kab mob hauv lub cev lossis cov kab mob hauv lub cev, nyob ntawm seb cov khoom noj muaj nyob, thiab tshwm sim raws li kev hloov ntawm txoj hnyuv plab thiab sab hauv mus rau hauv cov tswv tsev cell16. Thaum cov kab mob tau noj thiab tua los ntawm macrophages, qee qhov peptide fragments raug xa mus rau saum npoo ntawm antigen-presenting cell, raug encoded los ntawm qhov loj histocompatibility complex (MHC), chav kawm II. Qhov peptide-MHC II kev khi no txhawb nqa T CD4+ lymphocytes. Txawm li cas los xij, yog tias cov kab mob txiav txim siab nkag mus rau hauv lub xov tooj ntawm tes, nkag mus rau cytoplasm ntawm macrophage, kev sib txuas peptide nrog lwm hom MHC, chav kawm I, txhawb nqa T CD8+ lymphocyte ntau lawm17. Interestingly, CD4+ thiab CD8+ hlwb nthuav tawm tib tus qauv ntawm macrophages thoob plaws qhov kev sim, txawm tias sawv cev rau cov lus teb sib txawv. Txij li CD4+ thiab CD8+ hlwb feem ntau yog sawv cev rau T lymphocytes uas yog ib feem ntawm lub cev tiv thaiv kab mob, cov macrophages, yog ib feem ntawm lub cev tiv thaiv kab mob. Ntxiv rau qhov no, peb tau pom tias cov broilers nthuav tawm cov cim qhia tau zoo dua li cov hens, tau lees paub los ntawm kev tshawb fawb yav dhau los uas cov broilers tau sib tw nrog cov kab mob sib kis tau pom, piv txwv li, ntau qhov kev cuam tshuam hauv plab hnyuv thiab kab mob hauv lub cev11. Peb qhov kev tshawb pom qhia tias txoj hauv kev immunohistochemistry muab cov ntaub ntawv nthuav dav txog tus cwj pwm ntawm lub cev tiv thaiv kab mob ntawm ntau lub cev ntawm cov kab lag luam sib txawv thaum kis kab mob los ntawm Salmonella enterica serovars. Tsis tas li ntawd, cov pov thawj ntawm kev tshawb fawb tam sim no hais tias tshem tawm ob lub noob, txawm tias nyob hauv ntau hom Salmonella, ua rau cov kab mob uas ua rau lub cev tiv thaiv kab mob siab dua hauv cov tswv tsev, qhia tias cov kab mob tsis tau txo qis. Peb tuaj yeem txiav txim siab tias tej zaum Salmonella tuaj yeem nrhiav tau lwm txoj kev ciaj sia los ua tus kab mob ntau dua. Kev siv ttr thiab pdu operons nyob rau hauv consonance nrog cob thiab PRP operons tau qhia, nyob rau hauv lub yav dhau los txoj kev tshawb no, raws li tsim nyog rau anaerobic respiration16, ua rau peb ntseeg hais tias tsis yog tsuas yog yuav tsum tau tshem tawm ntau noob ntawm txhua operon18, tab sis peb kuj muaj. xav txog kev tshem tawm tag nrho cov teeb no, kom ncav cuag cov kab mob tsawg dua ntawm Salmonella enterica.

Cov ntaub ntawv thiab cov txheej txheem

Cov kev sim, ua raws li cov lus qhia thiab cov cai, tau pom zoo los ntawm Pawg Saib Xyuas Kev Ncaj Ncees ntawm Kev Siv Tsiaj ntawm Sao Paulo State University (CEUA/Unesp Process—006621/18; Lub Tsib Hlis 10, 2018), tau ua nyob rau hauv Avian Pathology Laboratory of Department of Pathology, Teriogenology, and One Health from the School of Agricultural and Veterinary Sciences, Sao Paulo State University (FCAV/Unesp), Jaboticabal, Brazil.

Cov kab mob kab mob thiab mutant tsim.

Cov kab mob uas siv ntawm no tau muab khaws cia rau hauv qhov nruab nrab cryoprotectant compounded los ntawm Lysogeny broth (LB; BD DifcoTM, USA) ntxiv nrog 30% ntawm glycerol (Merck, BR—H30402394 228) thiab cia rau hauv lub freezer ultra-freezer (− 80 degree) ntawm Avian Pathology Laboratory los ntawm FCAV/UNESP. Salmonella Enteritidis P125109 (tus lej nkag: AM933172) thiab Salmonella Typhimurium str. 9819 raug ntxias mus rau nalidixic acid- thiab spectinomycin-resistance (Nalr Spcr ) thiab lawv tau muab cov keeb kwm caj ces rau kev tsim cov kab mob mutant los ntawm Lambda-liab cov txheej txheem20 nrog kev hloov kho me me, piav qhia hauv Saraiva li al.11. Cov kab mob mutant tsim ntawm no tau txheeb xyuas cov ntawv nyeem raws li SEΔttrAΔpduA (Salmonella Enteritidis ∆ttrA∆pduA) thiab STMΔttrAΔpduA (Salmonella Typhimurium ∆ttrA∆due).

Hauv vivo kev sim.

Kev sim 1-Salmonella Enteritidis. Turty-rau 1-hnub qub menyuam qaib los ntawm txhua tus ntawm peb kab sib txawv (broiler, semi-heavy hens, thiab light laying hens), tag nrho ib puas thiab yim tsiaj, tau los ntawm kev lag luam hatcheries. Thaum tuaj txog, hauv qab ntawm cov thawv thauj khoom tau raug tshuaj xyuas kom paub meej tias Salmonella tsis pub muaj cov noog21, thiab cov tsiaj tau nyob hauv cov tawb hlau hauv chav acclimatized thiab tau txais tshuaj tua kab mob pub dawb thiab dej ad libitum. Ib qhov 24-h lub teeb tau raug xaiv rau thawj hnub kom ntseeg tau tias cov dej thiab cov khoom noj tau zoo, tom qab ntawd tau txais ib qho kev pom zoo 12-h hauv thawj lub lim tiam, txo mus rau 8 teev ntawm cov hnub tseem ceeb. Lub inoculum tau npaj raws li Berchieri Junior li al.22. Rau qhov no, cov kab lis kev cai khov tau raug inoculated hauv LB thiab incubated thaum hmo ntuj ntawm 37 degree hauv qab 150 rpm. Hnub tom qab, cov kab mob kab mob tau hloov mus rau hauv cov xov xwm tshiab thiab incubated rau 18 teev nyob rau hauv tib yam li yav tas los. Tom qab ntawd, 0.2 mL los ntawm cov kab lis kev cai uas muaj 108 colony-forming units ib mL (CFU / mL) tau hais lus inoculated los ntawm hlau gavage ncaj qha rau hauv cov noog qoob loo. Cuaj pawg tau tsim (A rau I) thiab muab faib ua ntu zus raws li cov kab sib txawv thiab hom kab mob (Table 7). Ntawm ib-, peb-, xya-, thiab 14-hnub tom qab kis kab mob (dpi), peb tus noog ib pab pawg txhua hnub, thaum sawv ntxov, tau euthanized los ntawm ncauj tsev menyuam dislocation los sau cov ntu nruab nrab ntawm caecal tonsils, caecum, thiab ileum, thiab ntu ntu ntawm lub siab sab laug lobe rau kev soj ntsuam ntxiv immunohistochemistry (IHC). Rau qhov no, cov qauv tau muab tso rau hauv n-Hexane pa (n-Hexano pa, Synth, Brazil) yav dhau los tso rau hauv cov kua nitrogen. Tam sim ntawd tom qab cov ntaub so ntswg khov, nws raug xa mus rau hauv 2 mL cryotube (Corning, USA) thiab txias hauv cov kua nitrogen. Tom qab ua piv txwv, cov ntaub so ntswg tau khaws cia ntawm -80 degree kom txog rau thaum cov txheej txheem rau IHC.

Kev sim 2-Salmonella Typhimurium. Qhov kev sim no tau ua raws li cov yam ntxwv uas tau hais los saum toj no hauv Kev sim 1. Tirty-rau qaib (1 hnub qub) tau muab faib ua cuaj pawg (A txog I) raws li lawv cov kab thiab hom kab mob (Table 7).

Table 7


Table 7. Tsim pab pawg raws li cov kab thiab kab sib txawv. SE∆ttrA∆pduA, Salmonella Enteritidis ∆ttrA∆pduA; STM∆ttrA∆pduA, Salmonella Typhimurium ∆ttrA∆pduA; wt-SE, Salmonella Enteritidis qus-hom; wt-STM, Salmonella Typhimurium qus-hom; NC, kev tswj tsis zoo.

Immunohistochemistry.

Cov ntaub so ntswg. Cov qauv sau tau raug xa mus los ntawm −80 degree mus rau cryostat (Leica CM1860, Leica Biosystems Nussloch GmbH, Lub teb chaws Yelemees) ntawm −22 degree qhov twg yog tus kheej thaiv hauv OCT compound (Tissue-Tek®, Sakura Finetek Europe BV, Netherlands) ib 30 feeb ua ntej mus rau 6 µm seem siv cov hniav uas tsis tshua muaj pov tseg (Leica 819, Leica Biosystems Nussloch GmbH, Lub Tebchaws Yelemees). Nws yog ib qho tseem ceeb uas cov seem tau ua tiav ntawm −22 degree, tshwj tsis yog daim siab, uas tau ua tiav ntawm -15 degree. Cov swb uas muaj peb qhov rov ua dua ntawm qhov ntu ntawm txhua lub cev ntawm lub cev tiv thaiv kab mob hauv lub cev tiv thaiv kab mob tau raug npaj, nrog thinning ntawm txhua qhov rov ua dua. Siv cov txhuam hniav, cov ntaub so ntswg txiav tau muab tso rau ntawm cov kab mob histological ua ntej kho nrog poly-l-lysine (Sigma-Aldrich, United Kingdom, Cat no. P4832) e silane (Sigma-Aldrich, USA. Cat no. 440574). Cov slides tau khaws cia ntawm -20 degree tom qab ntawd kom txog thaum IHC staining. Immune cell staining. Ua ntej, cov slides tau submerged nyob rau hauv 200 mL tub yees acetone (Acetone PA-ACS, Synth, Brazil) thiab incubated ntawm -20 degree rau 10 min. Tom qab ntawd, cov slides raug xa mus rau hauv cov av noo chamber (EasyPath®, Brazil) ntawm chav tsev kub rau 5 min kom qhuav cov qauv. Tom qab ntawd cov slides tau ntxuav nrog PBS thiab ib lub pas dej tau tso rau 5 feeb kom tsis txhob muaj cov ntaub so ntswg qhuav. Kaum, cov ntaub so ntswg tau submerged hauv 200 mL ntawm 4% H2O2 ib 10 min nyob rau hauv ib qho chaw tsaus thiab ntxuav dua nrog PBS. Qhov chaw nyob ib ncig ntawm cov ntaub so ntswg tau qhuav nrog cov ntawv nqus dej thiab cov qauv tau dhau los ntawm tus cwj mem hydrophobic (Dako Pen, Dako Denmark A / S, Denmark). Cov kauj ruam ntxuav tawm hauv lub pas dej tau rov ua dua li yav dhau los. Cov khoom siv biotin-free Mouse and Rabbit Specific HRP/DAB IHC Detection-Micropolymer (Abcam©, USA) tau siv los ua cov kab mob tiv thaiv kab mob, xaiv txoj kev Avidin-Biotin Streptavidin Peroxidase Complex (ABC). Rau qhov no, lub pas dej tau raug tshem tawm, thiab cov xim tsis muaj keeb kwm yav dhau los blocker reagent droplets tau ntxiv rau cov ntaub so ntswg. Cov slides tau khaws cia hauv cov av noo hauv qhov chaw tsaus rau 30 min, cov kauj ruam ntxuav tau rov ua dua, thiab 200 µL ntawm thawj cov tshuaj tiv thaiv kab mob (Nas Anti-Chicken CD4-UNLB; Mouse Anti-Chicken CD{{34} } UNLB; Mouse Anti-Chicken Monocyte/Macrophage-UNLB, Southern Biotech, USA) diluted hauv ib feem ntawm 1:200 (v/v) hauv cov tshuaj tiv thaiv kab mob diluent reagent (Antibody diluent, Abcam©, USA) tau ntxiv tom qab ntawd. Cov slides tau incubated ntawm 4 degree rau 18 teev. Hnub tom qab, cov slides raug ntxuav raws li yav tas los. Kaum, cov tee dej ntawm cov tshuaj tiv thaiv kab mob thib ob (Qhia Ntxiv, Abcam©, USA) tau ntxiv tom qab tshem tawm PBS ntau dhau, thiab cov av noo tau muab tso rau hauv qhov tsaus ntuj rau 30 min. Tom qab ntawd, poob ntawm 3,3′-diaminobenzidine (DAB Chromogen 50 ×, Abcam©, USA) tau diluted hauv 1 mL ntawm substrate (DAB Substrate, Abcam©, USA), uas ntim txaus rau peb slides, thiab ntxiv rau. cov ntaub so ntswg. Ib feeb tom qab, cov slides tau submerged hauv 200 mL dH2O rau 5 min. Tom qab ntawd, lawv tau pauv mus rau lub thawv yas uas muaj Harris hematoxylin (Êxodo Científca, Brazil) thiab raug tso tseg rau 1 min. Tom qab ntawd, cov slides tau ntxuav rau 10 min nyob rau hauv dej khiav ntawm qis siab. Cov slides raug xa mus rau cov cawv-xylene series (70% Alcohol, 90% Alcohol, 100% Alcohol, Xylene I, thiab Xylene II). Thaum kawg, cov ntawv npog tau muab tso rau ntawm cov swb tom qab ntxiv ib qho kev poob dej tsis muaj qhov nruab nrab (Entellan®, Merck, Brazil). Cov duab los ntawm cov ntaub so ntswg raug muab rho tawm, xaiv tsib qhov chaw saib random, siv lub tshuab ntsuas qhov muag (lens 400 ×) (Coleman®, qauv N-120) nrog lub koob yees duab digital adapter, rau kev txheeb xyuas ntxiv (Daim duab 1. ).

Cov ntaub ntawv tsom xam.


Desert ginseng—Improve immunity (2)

cistanche tubulosa- txhim kho lub cev tiv thaiv kab mob

Qhov feem pua ​​​​ntawm CD{{0}} thiab CD8+ hlwb thiab macrophages raug suav nrog siv Image-Pro Plus v.4.5.0.29 (MediaCybernetics, USA). Lawv raug ntsuas raws li qhov feem pua ​​​​ntawm cov txiaj ntsig ntawm lub cev tiv thaiv kab mob qhov chaw zoo / tag nrho cheeb tsam. Kev txheeb cais thiab cov duab tau ua tiav siv software GraphPad Prism v.8.0.1 rau macOS (GraphPad Sofware, La Jolla California, USA) thiab cov ntaub ntawv raug xa mus rau Variance Analysis (ANOVA) ua raws Bonferroni ntau qhov sib piv, xav txog qhov tseem ceeb ntawm qib qis dua 5% (P Tsawg dua lossis sib npaug li 0.05).

Figure 1


Daim duab 1. Ntu ntawm caecum ntawm broiler kis los ntawm Salmonella Typhimurium ∆ttrA∆pduA uas qhia txog kev tiv thaiv kab mob hauv macrophages, 7 hnub tom qab kis kab mob (× 400; Avidin-Biotin Streptavidin Peroxidase, counterstained nrog Hematoxylin).

Cov ntaub ntawv

1. Freitas Neto, OC, Penha Filho, RC & Barrow, PA Cov peev txheej ntawm tib neeg tsis-typhoid salmonellosis: Kev tshuaj xyuas. Braz. J. Poult. Sci. 12(1), 1–11. https://doi.org/10.1590/S1516-635X2010000100001 (2010).

2. Winter, SE et al. Lub plab o muab lub pa hluav taws xob ua pa rau Salmonella. Xwm Txheej 467, 426–429. https://doi.org/ 10.1038/nature09415 (2010).

3. Hinsley, AP & Berks, BC Qhov tshwj xeeb ntawm txoj kev ua pa cuam tshuam rau kev txo cov sulfur compounds los ntawm Salmonella enterica. Microbiology (Nyeem) 148, 3631–3638. https://doi.org/10.1099/00221287-148-11-3631 (2002).

4. Tiennimitr, P. et al. Kev mob plab hnyuv tso cai rau Salmonella siv ethanolamine los sib tw nrog microbiota. Proc. Natl. Acad. Sci. USA 108, 17480–17485. https://doi.org/10.1073/pnas.1107857108 (2011).

5. Staib, L. & Fuchs, TM Regulation of fucose and 1, 2-propanediol utilization by Salmonella enterica serovar Typhimurium. Pem hauv ntej. Microbiol. 6, 1116. https://doi.org/10.3389/fmicb.2015.01116 (2015). 6. Horswill, AR & Escalante-Semerena, JC Propionate catabolism nyob rau hauv Salmonella Typhimurium LT2: Ob chav hloov pauv sib txawv suav nrog prp locus ntawm 8.5 centisomes, prpR encodes ib tug tswv cuab ntawm sigma -54 tsev neeg ntawm activators BC, thiab ua tus operon. J. Bacteriol. 179, 928–940, ib. https://doi.org/10.1128/jb.179.3.928-940.1997 (1997).

7. Soria, MC, Soria, MA, Bueno, DJ & Terzolo, HR Sib piv ntawm 3 kab lis kev cai thiab PCR kev soj ntsuam rau Salmonella Gallinarum thiab Salmonella Pullorum nrhiav pom hauv cov nqaij qaib. Poult. Sci. 92, 1505–1515, ib. https://doi.org/10.3382/ps.2012-02926 (2013).

8. Van Immerseel, F. et al. Dynamics ntawm lub cev tiv thaiv kab mob infiltration nyob rau hauv caecal lamina propria ntawm qaib tom qab neonatal kab mob nrog ib tug kab mob Salmonella Enteritidis. Dev. Comp. Immunol. 26, 355–364 : kuv. https://doi.org/10.1016/s0145-305x(01)00084-2 (2002).

9. Montassier, HJ Fisiopatologia ua lub cev tiv thaiv kab mob. Hauv Doenças das Aves (eds AndreattiFilho, RL et al.) 467–489 (FACTA, 2020).

10. Nqe-Carter, M., Tingey, J., Bobik, TA & Roth, JR Lwm txoj kev txais hluav taws xob tetrathionate txhawb nqa B12-nyob ntawm kev loj hlob ntawm anaerobic ntawm Salmonella enterica serovar Typhimurium ntawm ethanolamine lossis 1,{{4} }propanediol. J. Bacteriol. 183, 2463–2475, ib. https://doi.org/10.1128/JB.183.8.{12}}.2001 (2001).

11. Saraiva, M. et al. Deciphering lub luag hauj lwm ntawm ttrA thiab pduA noob rau Salmonella enterica serovars nyob rau hauv ib tug qaib kab mob qauv. Avian Pathol. https://doi.org/10.1080/03079457.2021.1909703 (2021).

12. Beal, RK, Wigley, P., Powers, C., Barrow, PA & Smith, AL Cross-reactive cellular thiab humoral immune teb rau Salmonella enterica serovars Typhimurium thiab Enteritidis yog txuam nrog kev tiv thaiv rau heterologous rov sib tw. Vet. Immunol. Immunopathol. 114, 84–93 : kuv. https://doi.org/10.1016/j.vetimm.2006.07.011 (2006).

13. Lub caij ntuj no, SE & Bäumler, AJ Ib qho kev ua siab zoo: Txhawm rau sib tw nrog lub plab microbiota, Salmonella tsav nws lub tswv yim los muab cov pa hluav taws xob ua pa. Lub plab Microbes 2, 58–60. https://doi.org/10.4161/gmic.2.1.14911 (2011).

14. Rivera-Chavez, FI et al. Salmonella siv lub zog tsheb tavxij kom tau txais txiaj ntsig los ntawm kev mob plab hnyuv. PLoS Pathog. 9 Ib., 1003267. https://doi.org/10.1371/journal.ppat.1003267 (2013).

15. Khan, CM Te Dynamic Interactions ntawm Salmonella thiab microbiota, nyob rau hauv qhov nyuaj niche ntawm lub plab zom mov. Int. Sch. Res. Tsis yog. Xyoo 2014, 1–23. https://doi.org/10.1155/2014/846049 (2014).

16. Yoo, W., Kim, D., Yoon, H. & Ryu, S. Enzyme IIANtr tswj Salmonella ntxeem tau ntawm 1,2-propanediol thiab propionate catabolism. Sci. Rep. 7, 44827. https://doi.org/10.1038/srep44827 (2017).

17. Salyers, AA & Whitt, DD Host tiv thaiv cov kab mob kab mob: Tiv thaiv cov ntaub so ntswg thiab ntshav. Nyob rau hauv Bacterial Pathogenesis: A Molecular Approach (eds Salyers, AA & Whitt, DD) 16–29 (ASM Press, 1994).

18. Goes, V. et al. Salmonella Heidelberg sab-kauj ruam gene poob ntawm kev ua pa ua pa hauv tus qauv qaib. Microb kab mob. 171, 105725. https://doi.org/10.1016/j.micpath.2022.105725 (2022).

19. Barrow, PA, Hassan, JO & Berchieri, A. Jr. Kev txo qis hauv fecal excretion ntawm Salmonella Typhimurium strain F98 hauv qaib tau txhaj tshuaj tiv thaiv txoj sia thiab tua cov kab mob S. Typhimurium. Epidemiol. Kab mob. 104, 413–426. https://doi.org/10.1017/s095026880 0047439 (1990).

20. Datsenko, KA & Wanner, BL Ib-kauj ruam inactivation ntawm chromosomal noob nyob rau hauv Escherichia coli K-12 siv PCR cov khoom. Proc. Natl. Acad. Sci. USA 97, 6640–6645. https://doi.org/10.1073/pnas.1201632977 (2000).

21. Zancan, FB, Berchieri Junior, A., Fernandes, SA & Gama, NMSQ Salmonella spp. tshawb nrhiav hauv lub thawv thauj cov noog hnub qub. Braz. J. Microbiol. 31, 230–232 : kuv. https://doi.org/10.1590/S1517-83822000000300016 (2000).

22. Berchieri, A. Jr., Murphy, CK, Marston, K. & Barrow, PA Kev soj ntsuam ntawm kev pheej hmoo thiab kev xa mus ntsug ntawm Salmonella enterica serovars Pullorum thiab Gallinarum hauv qaib: Cov nyhuv ntawm cov kab mob thiab cov tswv tsev keeb kwm yav dhau los. Avian Pathol. 30, 221–231 : kuv. https://doi.org/10.1080/03079450120054631 (2001).

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