非洲猪瘟与典型猪瘟的诊断与防控

2018年8月,我国首例非洲猪瘟疫情在辽宁省沈阳市被确诊,该病的确诊以及原先存在的典型猪瘟必然会对我国养猪业健康发展造成冲击和影响。对非洲猪瘟和典型猪瘟的病原学、流行病学、诊断技术和防控4个方面进行了综述,以期为非洲猪瘟和典型猪瘟的有效综合防控提供参考。     

Antimicrobial‐resistant Enterobacteriaceae recovered from companion animal and livestock environments

Antimicrobial‐resistant bacteria represent an important concern impacting both veterinary medicine and public health. The rising prevalence of extended‐spectrum beta‐lactamase (ESBL), AmpC beta‐lactamase, carbapenemase (CRE) and fluoroquinolone‐resistant Enterobacteriaceae continually decreases the efficiency of clinically important antibiotics. Moreover, the potential for zoonotic transmission of antibiotic‐resistant enteric bacteria increases the risk to public health. Our objective was to estimate the prevalence of specific antibiotic‐resistant bacteria on human contact surfaces in various animal environments. Environmental surface samples were collected from companion animal shelters, private equine facilities, dairy farms, livestock auction markets and livestock areas of county fairs using electrostatic cloths. Samples were screened for Enterobacteriaceae expressing AmpC, ESBL, CRE or fluoroquinolone resistance using selective media. Livestock auction markets and county fairs had higher levels of bacteria expressing both cephalosporin and fluoroquinolone resistance than did equine, dairy, and companion animal environments. Equine facilities harboured more bacteria expressing cephalosporin resistance than companion animal shelters, but less fluoroquinolone resistance. The regular use of extended‐spectrum cephalosporins in livestock populations could account for the increased levels of cephalosporin resistance in livestock environments compared to companion animal and equine facilities. Human surfaces, as well as shared human and animal surfaces, were contaminated with resistant bacteria regardless of species environment. Detecting these bacteria on common human contact surfaces suggests that the environment can serve as a reservoir for the zoonotic transmission of antibiotic‐resistant bacteria and resistance genes. Identifying interventions to lower the prevalence of antibiotic‐resistant bacteria in animal environments will protect both animal and public health.     

High‐Level Ciprofloxacin‐Resistant Campylobacter jejuni Isolates Circulating in Humans and Animals in Incheon,Republic of Korea

Campylobacter jejuni is one of the major causative pathogens of outbreaks or sporadic cases of diarrhoeal diseases worldwide. In this study, we compared the phenotypic and genetic characteristics of C. jejuni isolates of human and food‐producing animal origins in Korea and examined the genetic relatedness between these two groups of isolates. Regardless of isolation source, all C. jejuni isolates harboured four virulence genes, cadF, cdtB, ciaB and racR, whereas the wlaN and virB11 genes were more frequently observed in human isolates. Antimicrobial susceptibility testing showed that the majority of C. jejuni isolates displayed high‐level resistance to fluoroquinolone (95.2%) or tetracycline (76.2%) antibiotics, and 12.4% of isolates exhibited multidrug resistance (more than three classes of antibiotics tested). Pulsed‐field gel electrophoresis (PFGE) of all Campylobacter isolates revealed 51 different SmaI‐PFGE patterns and six major clusters containing both human and animal isolates. These results indicate that genetically diverse strains of C. jejuni with antimicrobial drug‐resistance and virulence properties have prevailed in Incheon. Nevertheless, some particular populations continue to circulate within the community, providing the evidence for an epidemiological link of C. jejuni infections between humans and food‐producing animals. Therefore, the continued monitoring and surveillance of C. jejuni isolates of human and food‐producing animal origins are required for public health and food safety.     

温度对醉马草内生真菌共生体幼苗生长和生物碱产量的影响

研究了不同温度(8℃/5℃、15℃/12℃、22℃/15℃、28℃/25℃、35℃/28℃)处理对醉马草内生真菌共生体幼苗的形态指标、叶绿素、可溶性糖以及麦角酰胺和麦角新碱含量的影响,以期明确共生体幼苗生长和产碱的最适温度。结果表明,株高、根长和生物量均在22℃/15℃处理下达到最大,其中株高显著(P0.05)高于其它4个处理,根长显著(P0.05)高于28℃/25℃和35℃/28℃处理,生物量显著(P0.05)高于8℃/5℃、28℃/25℃和35℃/28℃处理,分蘖在15℃/12℃处达到最大,显著(P0.05)高于8℃/5℃、28℃/25℃和35℃/28℃处理。叶绿素在22℃/15℃处理下达到最大,显著(P0.05)高于8℃/5℃、15℃/12℃和35℃/28℃处理。可溶性糖在5个温度处理下差异不显著(P0.05)。在处理时间为15 d时,麦角酰胺和麦角新碱含量均在22℃/15℃处理下达到最大。综上所述,最适宜共生体幼苗生长、麦角酰胺和麦角新碱积累的处理温度是22℃/15℃。     

Contact and fumigant toxicity of oriental medicinal plant extracts against Dermanyssus gallinae (Acari: Dermanyssidae)

The acaricidal activity of methanolic extracts from 40 oriental medicinal plant species and a steam distillate of Cinnamomum camphora towards poultry house-collected adult Dermanyssus gallinae De Geer was examined using direct contact and vapour phase toxicity bioassays. Results were compared with those of 15 acaricides currently used. In filter paper contact toxicity bioassays using adult D. gallinae, C. camphora steam distillate (0.0051 mgcm(-2)) was the most toxic material, followed by extracts from Asarum sieboldii var. seoulens whole plant, Eugenia caryophyllata flower bud and Mentha arvensis var. piperascens whole plant (0.0063-0.0072 mgcm(-2)), based upon 24h LD(50) values. The acaricidal activity of these four plant preparations was almost comparable to that of profenofos (LD(50), 0.003 mgcm(-2)) but less effective than dichlorvos (LD(50), 0.0004 mgcm(-2)). The toxicity of Illicium verum fruit and Lysimachia davurica leaf extracts (0.09 mgcm(-2)) was almost comparable to that of benfuracarb, prothiofos, propoxur and fenthion (0.053-0.070mgcm(-2)). In vapour phase toxicity tests, these plant preparations were more effective in closed containers than in open ones, indicating that the mode of delivery of these plant extracts was largely a result of action in the vapour phase. Plants described herein merit further study as potential D. gallinae control agents.     

Expression of nitric oxide synthase isoforms in the testes of pigs

This study examined the expression of three isoforms of nitric oxide synthase (NOS) in the testes of pigs. Immunohistochemical studies demonstrated the presence of nNOS, eNOS and iNOS in interstitial cells, primary spermatocytes and spermatids. Positive immunoreactions for eNOS and iNOS were detected in peritubular myoid cells. Some vascular endothelial cells were positive for nNOS and eNOS. The expression of nitrotyrosine was detected in interstitial cells. In addition, the histochemical study revealed that all the interstitial cells were stained positively for NADPH-diaphorase, although some spermatids and vascular endothelial cells displayed moderate enzymatic activity. These findings suggest that three isoforms of NOS are expressed in the testis of pig and that they play important roles in the biology of interstitial cells that produce testosterone, as well as in spermatogenesis in the seminiferous tubules.     

Development of two immunochromatographic tests for the serodiagnosis of bovine babesiosis

In this study, we developed two immunochromatographic tests (ICTs), which are nitrocellulose membrane-based immunoassays for the convenient and rapid serodiagnosis of bovine babesiosis caused by Babesia bovis (BoICT) and Babesia bigemina (BiICT). The efficacy of two ICTs was evaluated using 13 positive sera from experimentally infected cattle with B. bovis or B. bigemina. Clear results showed that the BoICT and ELISA detected antibodies in sera collected from 14 to 93 days post-infection, while BiICT and ELISA detected from 13 to 274 days post-infection. In additon, non-infected cattle, Neospora caninum, and Cryptosporidium parvum were negative in two ICTs. To evaluate the field utility of the ICTs, we tested 186 field bovine sera collected from cattle living in Yanbian (China) and Mato Grosso do Sul (Brazil). The results of ICTs were compared to those of classical serodiagnostic methods, enzyme-linked immunosorbent assay (ELISA) and the indirect immunofluorescence assay (IFAT). The overall concordances of BoICT were determined as 92.5 and 90.3% when the results of ELISA and IFAT were set as the reference standards, respectively. In contrast, those of BiICT showed 96.8 and 92.5% relative to the results of standard ELISA and IFAT, respectively. Conventional and rapid diagnosic devices for bovine babesiosis may provide a valuable tool in clinical and field applications.     

Comparison of polymerase chain reaction methods for the detection of <Emphasis Type="Italic">Theileria equi</Emphasis> infection using whole blood compared with pre-extracted DNA samples as PCR templates

Rapid, efficient, and reproducible procedures for isolating DNA before PCR gene amplification are essential for the diagnosis of piroplasms. In this study, we evaluated the ease and reliability of detecting Theileria equi by PCR using pre-extracted DNA samples (by QIAamp DNA Mini Kit and phenol-chloroform methods) compared with blood spotted on FTA cards as PCR templates. Although minimal variations in limit of detection were observed among the methods compared, overall, the use of pre-extracted DNA samples and blood spotted on FTA cards had comparable detection limits. These results indicate that T. equi infection can be efficiently detected directly from FTA cards by PCR without the need for pre-extraction of DNA from blood samples.     

Molecular cloning and expression analysis of pig CD81

CD81, also known as TAPA-1 (target of antiproliferative antibody 1), is a member of the tetraspanin family of proteins and a component of the B cell co-receptor complex. Several studies have shown that CD81 plays significant roles in a variety of immune responses, including activation of B cells and T cells. In this study, we cloned pig Cd81 cDNA using RT-PCR coupled with rapid amplification of cDNA ends (RACE)-PCR and determined the complete cDNA sequence of pig Cd81. Pig Cd81 cDNA contains an open reading frame (711 bp) encoding 236 amino acids. The identity of pig CD81 with those of human, cattle, rat, and mouse are 90.30%, 92.26%, 86.22%, and 86.22%, respectively. Alignment of the CD81 amino acid sequence with those of mammalian species showed that the large extracellular loop (LEL) is the most divergent, whereas other domains are largely conserved. Pig Cd81 mRNA was detected by RT-PCR in a broad range of tissues, including lymphoid tissues as well as nonlymphoid tissues, indicated variety of cellular functions of CD81 in most pig tissues. Flow cytometry analyses demonstrated that human CD81 antibody recognizes a pig CD81 on the cell surface. Further, immunohistochemistry analysis using human CD81 antibody on pig spleen was revealed that CD81 expression is widely diffused in spleen tissue. Future study will be focused on defining the functional role of CD81 during the course of pig infectious diseases.