Fasciola spp. infection and molecular identification in cattle and sheep in Wushen Banner, Inner Mongolia

doi:10.12140/j.issn.1000-7423.2024.01.005

Abstract

Abstract:

Objective To investigate the endemic status of Fasciola spp. infection in cattle and sheep in Wushen Banner of Inner Mongolia. Methods From August 2022 to March 2023, blood and fecal samples from cattle and sheep were collected from Galutu, Sulide Sumu, Wulantaolegai, Wushenzhao and Tuke towns in Wushen Banner to detect serum Fasciola hepatica antibodies by using indirect ELISA. Cattle and sheep feces samples corresponding to some serum antibody detection samples were randomly selected, in which the parasite eggs were qualitatively examined by egg sedimentation method. The consistency test was performed for the findings from serum and fecal examination. In April 2023, Fasciola spp. adult worms and eggs were collected from the carcasses livers and bile of cattle and sheep that died of Fasciola spp. infection in Galutu, Sulidesumu and Tuke towns. The DNA of the worms and eggs was extracted to amplify ribosomal transcriptional spacer sequence 2 (ITS2) by PCR and sequenced. Sequence alignment was then performed by BLAST with NCBI database. The phylogenetic tree was constructed by the neighbour-joining method. SPSS 25.0 software was used for statistical analysis, and the chi-square test was used to compare the positive rates in different regions. Results A total of 825 serum samples were collected from cattle and sheep, in which 295 samples were positive for serological antibodies, with a total sero-positive rate of 35.8% (295/835), of which the sero-positive rates of sheep and cattle were 34.7% (227/655) and 40.0% (68/170), respectively. The sero-positive rate of sheep was the highest at 81.8% (45/55) in Sulide Sumu, and the lowest at 22.0% (27/123) in Wushenzhao. There was a significant difference in the sero-positive rate of antibodies in different regions (χ² = 73.93, P < 0.05). The sero-positive in bovine was the highest at 65.1% (28/43) in Garutu and lowest at 19.2% (5/26) in Tuke. There was a significant difference in the sero-positive rate of antibodies in different regions (χ² = 21.50, P < 0.05). A total of 88 fecal samples (59 sheep fecal samples and 29 bovine fecal samples) were detected, among which 1 sheep and 5 bovine fecal samples were positive for F. hepatica eggs, and 4 bovine fecal samples were positive for Paramphistomum eggs. Among the 88 samples, the positive rate of anti-F. hepatica antibody was 29.5% (26/88), and the positive rate of F. hepatica eggs was 6.8% (6/88). The kappa value was 0.086, indicating weak consistency between the two methods. Adult worms and eggs were collected from 2 dead cattle and adult worms from 2 dead sheep. The DNA were extracted from 5 adult worms and 2 eggs samples. A total of 7 bands of approximately 550 bp were obtained by PCR amplification for sequencing. BLAST alignment analysis showed that the identities of 4 obtained sequences were 92.97%, 96.62%, 95.74% and 95.91% with the ITS2 sequences (GenBank: HQ700438) of F. gigantica from China, respectively, and identified as F. gigantica. The identities of the other three obtained sequences were 95.69%, 99.80% and 99.23% with the ITS2 sequences (GenBank: JF496717) of F. hepatica from China, respectively, and identified as F. hepatica. The phylogenetic tree showed that the F. hepatica identified in this study clustered on the same branch with the F. hepatica from China (GenBank: JF496717), Kenya (GenBank: MZ396926) and Egypt (GenBank: MW620063). The F. gigantica identified in this study clustered in the same branch with the F. gigantica from China (GenBank: HQ700438) and India （GenBank: OL691113）. Conclusion F. hepatica and F. gigantica infections are present in both cattle and sheep in Wushenqi area of Inner Mongolia, and the infection rate of Fasciola spp. was relatively high.

Key words: Fasciola spp., Fasciola hepatica, Fasciola gigantica, Serum antibody, ELISA, Molecular identification, Cattle and sheep, Inner Mongolia

CLC Number:

R383.26

LI Na, ZANG Dare, WURI Lige, ALATENG Burigude, HAI Ying, HASI Surong. Fasciola spp. infection and molecular identification in cattle and sheep in Wushen Banner, Inner Mongolia[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2024, 42(1): 36-41.

Figures/Tables 5

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References 25

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采样点 Site of collection	羊 Sheep			牛 Bovine			合计 Total
采样点 Site of collection	血样数/份 No. blood sample	阳性数/份 No. positive	阳性率/% Positive rate/%	血样数/份 No. blood sample	阳性数/份 No. positive	阳性率/% Positive rate/%	血样数/份 No. blood sample	阳性数/份 No. positive	阳性率/% Positive rate/%
嘎鲁图 Galutu	244	82	33.6	43	28	65.1	287	110	38.3
苏力德苏木 Sulidesumu	55	45	81.8	35	17	48.6	90	62	68.9
乌兰陶勒盖 Wulantaolegai	116	38	32.8	33	9	27.3	149	47	31.5
乌审召 Wushenzhao	123	27	22.0	33	9	27.3	156	36	23.1
图克 Tuke	117	35	29.9	26	5	19.2	143	40	28.0
合计 Total	655	227	34.7	170	68	40.0	825	295	35.8

粪便虫卵检查 Fecal egg examination	血清抗体检测 Serum antibody detection		合计 Total
粪便虫卵检查 Fecal egg examination	阳性 Positive	阴性 Negative	合计 Total
阳性 Positive	3	3	6
阴性 Negative	23	59	82
合计 Total	26	62	88