Aims: To develop a PCR‐based assay to detect Prototheca zopfii (P. zopfii) and its mastitis‐related subtype (genotype 2) directly from milk samples. Methods and Results: The DNA extraction method herein is based on the lysing properties of chemical agents, mechanical grinding and DNA‐binding properties of silica particles; this method was developed to rapidly extract DNA directly from P. zopfii in bovine milk. Two pairs of primers specific for P. zopfii and genotype 2 were used in the duplex PCR, and a sensitivity test showed that the detection level was 5 × 102 colony‐forming units (CFU) ml−1 for P. zopfii and 5 × 103 CFU ml−1 for genotype 2. Furthermore, a practical survey of 23 milk samples showed that the assay produced results that were in accordance with those obtained by the conventional microbiology method. Conclusions: The DNA extraction method is effective in isolating sufficient quantities of DNA from P. zopfii in milk for PCR analysis. The PCR assay is economical, sensitive and more rapid than the conventional culture method. Significance and Impact of the Study: The assay could be used as an alternative method for the rapid the detection of bovine mastitis resulting from P. zopfii genotype 2.
The biodegradation of toluene and benzene in a biofilter using cylindrical activated carbon as the filler materials was studied. Three gas low rates, i.e. 0.25, 0.50 and 0.75 m3/h, corresponding to empty bed gas residence of 75, 37.5 and 25 s, respectively, and total organic load lower than 400 g/m3 x h were tested. The biofilter proved to be highly efficient in biodegradation of toluene and benzene, and toluene was more easily degraded than benzene. When each inlet load was lower than 150 g/m3 x h, removal rate increased with inlet load and reached a maximum, which was 150 and 120 g/m(-3) x h for toluene and benzene, respectively. For inlet load higher than the maximum removal capacity conditions, the removal rate decreased with inlet load. Carbon dioxide concentration profile through the biofilter revealed that the mass ratios of carbon dioxide produced to the toluene and benzene removed were 2.15 g(CO2)/g(toluene) and 1.67 g(CO2)/g(benzene), which furthermore, confirmed the biodegradation performance in biofilter. The observation of biotic community demonstrated that the microbes consisted of bacillus, spore bacillus and fungi, of which the spore bacillus was dominant.
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