A novel and safe method has been reported for wastewater sludge treatment, the first step of which is H2O2 pretreatment at 150 cm3kg for 80 min. In this step, the majority of organic substrate was degraded, and the shear stress diminished significantly. In the second step, Aspergillus oryzae further decomposed the organic substrate of sludge for 8 days, and the activities of manganese peroxidase and lignin peroxidase kept increasing. By the methods, the residual chemical oxygen demand (COD) in the sludge was much lower than that in the sludge treated with high doses of H2O2. These results indicate that the present study provides a feasible method to safely dispose sludge from the wastewater treatment plant.
In this study, two pairs of oligonucleotide primers were designed according to the nucleotide sequence of the internal transcribed spacers (ITSs) of Babesia bigemina and B. bovis isolates from China. The primers were used in a multiplex PCR to detect parasite DNA in blood samples from cattle. There was no cross reactions with B. ovata, B. major, B. sp. Kashi, Theileria annulata, T. sergenti, T. sinensis or normal bovine DNA. The sensitivity of multiplex PCR assay was 1 pg and 10 pg DNA for B. bigemina and B. bovis, respectively. A total of 260 field blood samples collected from cattle in five provinces of China were analyzed by multiplex PCR and light microscopy. PCR testing revealed that 7.3% (19/260) and 5.8% (15/260) of cattle were positive for B. bigemina and B. bovis and 1.2% (3/260) of cattle were co-infected with B. bigemina and B. bovis. Using light microscopy, 2.3% (6/260) and 1.5% (4/260) of cattle were infected by B. bigemina and B. bovis, respectively, and no co-infection was found. The results showed that the multiplex PCR developed in the present study could be an alternative diagnostic tool for the detection of B. bovis and B. bigemina infection in cattle.