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ABSTRACT: Lamb diarrhea seriously restricts the development of the sheep industry. Infectious pathogens often cause diarrhea, and E. coli isolates are highly distributed in infectious diarrhea. The study aimed to investigate the prevalence of pathogenic E. coli in diarrhea lambs and determine the distribution of virulence genes, antibiotic resistance genes, and antibiotic resistance. One hundred seventy-eight E. coli isolates were isolated from the feces of 204 diarrhea lambs. The virulence genes mdh, hlyF, iss, ompA, fimC, iucD, st, lt, stx1, stx2, and antibiotic resistance genes including tetA, tetB, aac (6′)-II, blaCMY-2, qnr, aadA1, sul1, blaTEM, blaCTX-M were detected by polymerase chain reaction (PCR). Antibiotic resistance was determined by Kirby-Bauer Disc Diffusion method. There were 109 (61.24%, 109/178) enterotoxigenic E. coli (ETEC), 119 (66.85%, 119/178) Shiga toxin-producing E. coli (STEC), and 95 (53.37%, 95/178) hybrid STEC/ETEC isolates. The highest prevalent virulence genes were ompA (80.90%, 144/178) and fimC (67.98%, 121/178), and the lowest was iucD (8.99%, 16/178). The most commonly detected antibiotic resistance genes were tetA/tetB (92.70%, 165/178), qnr (75.84%, 135/178), and sul1 (62.92%, 112/178), no blaTEM or blaCTX-M genes were detected. All isolates had high antibiotic resistance to lincomycin (96.63%, 172/178), tetracycline (88.76%, 158/178), and co-trimoxazole (80.34%, 143/178), and the multidrug resistant (MDR) rate reached 93.82% (167/178). The high prevalence of ETEC and STEC indicates that E. coli is one of the critical pathogenic agents leading to diarrhea in lambs in the region, and its high antibiotic resistance, especially MDR, should be brought to our attention. ABSTRACT industry E seventyeight seventy eight 20 mdh hlyF iss st lt stx1 stx stx2 tetA tetB 6′II, 6II II 6′ II, 6 (6′)-II blaCMY2, blaCMY2 blaCMY 2, 2 blaCMY-2 aadA1 aadA sul blaCTXM blaCTX M PCR. PCR . (PCR) KirbyBauer Kirby Bauer method 10 61.24%, 6124 61 24 (61.24% 109/178 109178 178 ETEC, , (ETEC) 11 66.85%, 6685 66 85 (66.85% 119/178 119178 toxinproducing toxin producing STEC, (STEC) 9 53.37%, 5337 53 37 (53.37% 95/178 95178 STECETEC 80.90%, 8090 80 90 (80.90% 144/178 144178 144 67.98%, 6798 67 98 (67.98% 121/178, 121178 121/178 121 121/178) 8.99%, 899 8 99 (8.99% 16/178. 16178 16/178 16 16/178) tetAtetB 92.70%, 9270 92 70 (92.70% 165/178, 165178 165/178 165 165/178) 75.84%, 7584 75 84 (75.84% 135/178, 135178 135/178 135 135/178) 62.92%, 6292 62 (62.92% 112/178, 112178 112/178 112 112/178) 96.63%, 9663 96 63 (96.63% 172/178, 172178 172/178 172 172/178) 88.76%, 8876 88 76 (88.76% 158/178, 158178 158/178 158 158/178) cotrimoxazole co trimoxazole 80.34%, 8034 34 (80.34% 143/178, 143178 143/178 143 143/178) MDR (MDR 9382 93 82 93.82 167/178. 167178 167/178 167 (167/178) region attention 6′II blaCMY- (PCR 1 61.24% 612 (61.24 109/17 10917 17 (ETEC 66.85% 668 (66.85 119/17 11917 (STEC 53.37% 533 5 3 (53.37 95/17 9517 80.90% 809 (80.90 144/17 14417 14 67.98% 679 (67.98 12117 121/17 12 8.99% 89 (8.99 1617 16/17 92.70% 927 7 (92.70 16517 165/17 75.84% 758 (75.84 13517 135/17 13 62.92% 629 (62.92 11217 112/17 96.63% 966 (96.63 17217 172/17 88.76% 887 (88.76 15817 158/17 15 80.34% 803 (80.34 14317 143/17 938 93.8 16717 167/17 (167/178 61.24 (61.2 109/1 1091 66.85 (66.8 119/1 1191 53.37 (53.3 95/1 951 80.90 (80.9 144/1 1441 67.98 (67.9 1211 121/1 8.99 (8.9 161 16/1 92.70 (92.7 1651 165/1 75.84 (75.8 1351 135/1 62.92 (62.9 1121 112/1 96.63 (96.6 1721 172/1 88.76 (88.7 1581 158/1 80.34 (80.3 1431 143/1 93. 1671 167/1 (167/17 61.2 (61. 109/ 66.8 (66. 119/ 53.3 (53. 95/ 80.9 (80. 144/ 67.9 (67. 121/ 8.9 (8. 16/ 92.7 (92. 165/ 75.8 (75. 135/ 62.9 (62. 112/ 96.6 (96. 172/ 88.7 (88. 158/ 80.3 143/ 167/ (167/1 61. (61 66. (66 53. (53 80. (80 67. (67 8. (8 92. (92 75. (75 62. (62 96. (96 88. (88 (167/ (6 (5 ( (9 (7 (167 (16 (1