Reproductive incompatibility between the B and Q biotypes of the whitefly Bemisia tabaci in China: genetic and behavioural evidence.

The B and Q 'biotypes' of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) have been invading many parts of the world and causing severe damage to a range of crops. Recent phylogenetic analyses indicate that B and Q are cryptic species within the B. tabaci species complex. Although various attempts have been made to examine the reproductive compatibility between B and Q, few studies have tested the fertility of the F1 females and so the extent of possible gene flow remains unclear. In this study, we conducted a series of crossing experiments and behavioural observations to examine in detail the reproductive compatibility between the B and Q biotypes collected from Zhejiang, China, a region recently invaded by these whiteflies. Crossing experiments between the two biotypes using either single-pairs or small groups demonstrated that proportions of females in the F1 progeny were only 0-2% in the inter-biotype crosses compared to 58-68% in the intra-biotype treatments. Furthermore, all inter-biotype F1 females were sterile. Continuous video observations showed that B and Q adults very rarely copulated, and copulation occurred only when adults of opposite sex from different biotypes were enclosed in dense cohorts for a relatively long period of time. These data show that the B and Q biotypes examined in this study are completely isolated in reproduction. The isolation was due to mainly a copulation barrier, but post-copulation barriers were also involved.

An indirect ELISA for serodiagnosis of cattle footrot caused by Fusobacterium necrophorum.

A serodiagnostic ELISA (rL-ELISA) using recombinant truncated leukotoxin protein PL2 (aa 311-644) of Fusobacterium necrophorum as antigen was developed for detection of antibodies against F. necrophorum from cattle footrot. In rL-ELISA, the recombinant diagnostic antigen showed no cross-reaction with antisera against bovine foot and mouth disease virus, bovine rhinotracheitis virus, bovine viral diarrhea virus, bovine rotavirus type A, bovine Escherichia coli, and bovine Salmonella. The rL-ELISA could confirm the existence of antibodies against F. necrophorum at day 7 after infection. Detection of the field samples indicated relative sensitivity of rL-ELISA to nL-ELISA using the purified native leukotoxin A as antigen was 96.43%, and relative specificity of rL-ELISA to nL-ELISA was 94.26%. These data demonstrated the rL-ELISA would have a potential use for early diagnosis of cattle footrot caused by F. necrophorum.

Identification of three immunodominant regions on leukotoxin protein of Fusobacterium necrophorum.

To analyze immunodominant regions of leukotoxin protein of Fusobacterium necrophorum strain H05, a series of truncated forms of leukotoxin gene were expressed in Escherichia coli using the vector pGEX-6p-1 or pPROEX HTa. The results of SDS-PAGE showed the truncated forms PL1, PL2, PL4, and PL5 were expressed in Escherichia coli using the vector pGEX-6p-1, and the truncated forms PL3 was expressed in Escherichia coli using the vector pPROEX HTa. These recombinant proteins were able to react with antisera against Fusobacterium necrophorum strain A25. In five recombinant proteins, the recombinant proteins PL1, PL3 and PL4 as vaccine were able to elicit formation of the better protective effects on mice against infection of Fusobacterium necrophorum strain A25.

Spike protein region (aa 636789) of porcine epidemic diarrhea virus is essential for induction of neutralizing antibodies.

To analyze antigenic structure of the spike (S) protein of Porcine epidemic diarrhea virus (PEDV), the gene encoding its major immunodominant region S1 was amplified by PCR. We prepared four truncated S1 proteins spanning the entire S1 domain fused to GST protein. To identify the most important antigenic region of S1, the truncated S1-GST fusion proteins were examined for their ability to react with immune serum against PEDV and to elicit the formation of neutralization antibodies in immunized animals. We found that the region of S1 signed as S1D (aa 636789) was able to react with PEDV antiserum and to elicit formation of neutralization antibodies in mice. Moreover, the immune serum against S1D showed the binding ability to the native S protein of PEDV.

Isolation and molecular characterization of a porcine teschovirus 1 isolate from China.

Porcine teschovirus 1 (PTV-1) (Swine/CH/IMH/03) was isolated from piglets in a farm in Inner Mongolia Province, P.R. China. It was confirmed by electron microscopy, RT-PCR, and sequencing. Comparison of the sequences of the amino acid and nucleotides and phylogenetic analysis of the polyprotein showed that PTV Swine/CH/IMH/03 strain is PTV-1. The isolated virus has closest relationship with Talfan strain, they shared 98.9% and 99.5% homology of amino acids and nucleotides, respectively, in the ORF of polyprotein. To our knowledge, this is the first report about isolation and identification of a PTV in China.