ABSTRACT
Objective: To determine the infection rates and subtype distributions of Blastocystis in pigs, cattle, and goats in Hainan Province, China. Methods: From September 2018 to May 2019, a total of 659 fresh fecal samples were collected from 188 pigs, 341 goats, and 130 cattles farmed in cities of Hainan Province. All the samples were analyzed for the presence of Blastocystis by amplifying the SSU rRNA gene. Phylogenetic analysis was performed by constructing a neighborjoining tree using the program Mega X. Results: Thirty-one percent (204/659) of the animals were confirmed to be infected with Blastocystis, with 42.2% (144/341) of the goats, 30.8% (40/130) of the cattle, and 10.6% (20/188) of the pigs. Twelve subtypes of Blastocystis were identified, including ST10 (n=114), ST5 (n=37), ST26 (n=16), ST24 (n=8), ST14 (n=8), ST21 (n=8), ST23 (n=4), ST25 (n=4), ST1 (n=2), ST2 (n=1), ST4 (n=1), and ST12 (n=1). There were 11 (ST1, ST2, ST4, ST5, ST10, ST12, ST14, ST21, ST23, ST24, and ST26) and six subtypes (ST10, ST21, and STs23- ST26) in goats and cattle, respectively. All the pig-derived Blastocystis isolates belonged to ST5. A total of 37 representative sequences were obtained from the 12 subtypes of Blastocystis, including 14 sequences that have been described previously and 23 novel sequences were identified which were composed of ST10 (n=9), ST5 (n=8), ST21 (n=3), ST26 (n=2), and ST14 (n=1). Conclusions: This is the first study to identify the presence of Blastocystis in pigs, cattle, and goats from Hainan Province, China, and provides baseline data useful for controlling and preventing Blastocystis infection in farm communities. Pigs and goats appear to be the major reservoirs of potential zoonotic infections with ST1, ST2, ST4, ST5, and ST12 and zoonotic infections in pigs and goats with those subtypes should be considered potential public health threats. The new ST sequences of Blastocystis identified here provide novel insights into the genotypic variation in Blastocystis.
ABSTRACT
The aim of the present study is to identify and characterize lucerne lines resistance to weevil infestation. After three years of field screening for resistance to weevil infestation, 13 lines of lucerne were selected to assess the genotypic variations for lucerne weevil (Hypera postica Gyll.) at biochemical and molecular levels. Total phenols varied from 0.15 to 0.91 mg g-1 (DM) in these genotypes. The highest trypsin (11.11 unit mg-1 protein) and chymotrypsin (93.0 unit mg-1 protein) inhibitors activities were recorded in G-1-02 and B-4-03 lines respectively, whereas highest a-amylases inhibitor activity (14.2 unit mg-1 protein) in C-6-01. Zymogram patterns for trypsin inhibitor activity showed quantitative variations among the lines. In total 262 DNA fragments were generated when 45 deca-mer random primers were employed. Genetic variation in terms of genetic distance ranged from 0.65 to 0.85. Sequential Agglomerative Hierarchical and Nested (SAHN) clustering using the Un-weighted Pair Group Method with Arithmetic mean (UPGMA) algorithm yielded two clusters (cluster I and II) which converged at 72% similarity level. Cluster I contained most of the lines having low level of weevil infestation. High bootstrap values (>40) indicated the significance of nodes embodied in these two clusters. However, SDSPAGE analysis of the leaf proteins of these 13 lines showed no major variations except minor difference in the protein bands of molecular weights between 14 to 20 kD.