图1 分离株在产ESBL显色培养基(A)、麦康凯培养基(B)、营养肉汤培养基(C)上的菌落形态

Fig.1 Colony morphology of isolate on ESBL chromogenic medium (A), MacConkey medium (B) and nutrient broth medium (C)

图2 部分分离株16S rRNA基因序列PCR扩增结果 M,DL2000 DNA marker;1~2,阳性样品;3,阴性对照。

Fig.2 PCR amplification results of 16S rRNA gene sequences of isolates M, DL2000 DNA marker;1-2, Positive samples; 3, Negative control.

图3 16株分离株sul2基因PCR扩增结果 M,DL2000 DNA marker;泳道1~16,sul2基因阳性样品;17,阴性对照。

Fig.3 PCR amplification results of sul2 gene in 16 isolates M, DL2000 DNA marker; Lanes 1-16,sul2 gene positive samples; 17, Negative control.

图4 16株分离株TEM基因PCR扩增结果 M,DL2000 DNA marker;泳道1~3、5~6、8~10、13~15,TEM基因阳性样品;泳道4、7、11、12、16,阴性样品;17,阴性对照。

Fig.4 PCR amplification results of TEM gene in 16 isolates M, DL2000 DNA marker; Lanes 1-3, 5-6, 8-10, 13-15, TEM gene positive samples; Lanes 4, 7, 11, 12, 16, Negative samples; 17, Negative control.

图5 16株分离株荚膜血清型K5基因PCR扩增结果 M,DL2000 DNA marker;泳道10、13、15,荚膜血清型K5基因阳性样品;泳道1~9、11~12、14、16,阴性样品;17,阴性对照。

Fig.5 PCR amplification results of capsular serotype K5 gene in 16 isolates M, DL2000 DNA marker;Lanes 10, 13, 15, Capsular serotype K5 gene positive samples; Lanes 1-9, 11-12, 14, 16, Negative samples; 17, Negative control.

图6 16株分离株毒力基因mrkD基因序列PCR扩增结果 M,DL2000 DNA marker;泳道2~16,毒力基因mrkD基因阳性样品;泳道1,阴性样品;17,阴性对照。

Fig.6 PCR amplification results of virulence gene mrkD in 16 isolates M, DL2000 DNA marker; Lanes 2-16,Positive samples of virulence gene mrkD; Lane 1,Negative sample; 17,Negative control.

图7 小鼠整体剖检情况 A,空白对照组小鼠剖检整体情况无异常;B,弱毒力菌株组小鼠剖检整体情况为胆囊肿大,肠道积气积液、肿大;C,强毒力菌株组小鼠整体剖检情况为明显胆囊肿大,肠道积气积液、肿大。

Fig.7 Whole dissection of mice A,No abnormalities in mice under control; B,Gallbladder enlargement, gas accumulation in the intestine, and swelling in mice with weakly virulent strain; C,Obvious gallbladder enlargement, gas accumulation in the intestine, and enlargement in mice with strongly virulent strain.

图8 正常小鼠,以及攻毒小鼠各器官形态 A,空白对照组小鼠的肺无异常;A1,弱毒力菌株组小鼠肺充血肿大;A2,强毒力菌株组小鼠肺明显充血肿大。B,空白对照组小鼠脾无异常;B1,弱毒力菌株组小鼠脾淤血肿大;B2,强毒力菌株组小鼠脾明显淤血肿大。C,空白对照组小鼠肝无异常;C1,弱毒力菌株组小鼠肝淤血肿大;C2,强毒力菌株组小鼠肝明显淤血肿大。D,空白对照组小鼠肾无异常;D1,弱毒力菌株组小鼠肾充血肿大;D2,强毒力菌株组小鼠肾明显充血肿大。

Fig.8 Morphology of organs in normal mice, as well as virulent mice A, No abnormalities in the lung of mice under control; A1, Congested and enlarged lung in the mice with weakly virulent strain; A2, Obviously congested and enlarged lung in the mice with strongly virulent strain. B, No abnormalities in the spleen of mice under control; B1, Congested and enlarged spleen in the mice with weakly virulent strain; B2, Obviously congested and enlarged spleen in the mice with strongly virulent strain. C, No abnormalities in the liver of mice under control; C1, Congested and enlarged liver in the mice with weakly virulent strain; C2, Obvious congested and enlarged liver in the mice with strongly virulent strain. D, No abnormalities in the kidney of mice under control; D1, Congestion and enlarged kidney in the mice with weakly virulent strain; D2, Obvious congested and enlarged kidney in the mice with strongly virulent strain.

图9 攻毒小鼠组织病理学变化 A,肝淤血(100×);B,肝胞核固缩,中性粒细胞(400×);C,脾被膜增厚(100×);D,脾细胞坏死形成“星空现象”(400×);E,肺泡膈轻微增厚(100×);F,肺中性粒细胞增多(400×);G,肾小球肿大,絮状物沉积(100×);H,肾小球肿大(400×)。

Fig.9 Histopathological changes in virulent mice A, Liver congestion (100×);B, Hepatic nucleopyknosis, neutrophils (400×); C, Thickening of the splenic capsule (100×); D, Necrosis of splenocytes “Starry sky phenomenon” (400×); E, Slightly thickened alveolar diaphragm (100×); F, Lung neutrophils increased (400×); G, Glomerular enlargement, flocculent sedimentation (100×); H, Glomerulus enlargement(400×).