The nature and consequences of co-infections in tilapia: A review

Co-infections commonly arise when two or multiple different pathogens infect the same host, either as simultaneous or as secondary concurrent infection. This potentiates their pathogenic effects and leads to serious negative consequences on the exposed host. Numerous studies on the occurrence of the bacterial, parasitic, fungal and viral co-infections were conducted in various tilapia species. Co-infections have been associated with serious negative impacts on susceptible fish because they increase the fish susceptibility to diseases and the likelihood of outbreaks in the affected fish. Co-infections can alter the disease course and increase the severity of disease through synergistic and, more rarely, antagonistic interactions. In this review, reports on the synergistic co-infections and their impacts on the affected tilapia species are highlighted. Additionally, their pathogenic mechanisms are briefly discussed. Tilapia producers should be aware of the possible occurrence of co-infections and their effects on the affected tilapia species and in particular of the clinical signs and course of the disease. To date, there is still limited information regarding the pathogenicity mechanisms and pathogen interactions during these co-infections. This is generally due to low awareness regarding co-infections, and in many cases, a dominant pathogen is perceived to be of vital importance and hence becomes the target of treatment while the treatment of the co-infectious agents is neglected. This review article aimed at raising awareness regarding co-infections and helping researchers and fish health specialists pay greater attention to these natural cases, leading to increased research and more consistent diagnosis of co-infectious outbreaks in order to improve control strategies to protect tilapia when infected with multiple pathogens.     

规模化畜禽养殖场粪便中多重耐药菌分离鉴定及其耐药特征

为了解禽畜粪便中多重耐药菌的污染特征,本研究对鸡粪、牛粪、猪粪和有机肥这4种不同样品中多重耐药菌进行了计数分析,并对不同来源的多重耐药菌株进行了分离和纯化,进一步开展了基于16S rDNA序列比对的分子生物学鉴定以确定其种属地位,以及基于药敏试验的耐药性特征分析。结果表明:不同养殖动物粪便中四环素、恩诺沙星、磺胺甲恶唑和泰乐菌素4种抗生素多重耐药菌的绝对数量和相对数量排序均为鸡粪>猪粪>牛粪,粪源有机肥中可培养的多重耐药菌的绝对数量和相对数量在堆肥后均有所下降。通过对耐药菌株鉴定和分类学分析,发现禽畜粪便中多耐药菌的菌门集中分布在Proteobacteria、Firmicutes和Actinobacteria,多重耐药菌的优势菌属为Escherichia、Corynebacterium、Kurthia;而有机肥样品中多重耐药菌的优势菌属为Staphylococcus、Glutamicibacter。菌株的药敏试验表明,3类动物粪污中的多重耐药菌都对红霉素、四环素有着较高的耐药率,均高于80%,而对阿米卡星这种抗生素表现出较好的敏感性,其耐药率低于30%。通过对Ⅰ、Ⅱ类整合子基因盒的扩增,发现PCR产物的大小从0.8 kb到1.8 kb不等,基因盒ad A2、dfr A17、dfr A1及sat2在养殖场粪污中检出率均较高。     

草地贪夜蛾对小麦的为害风险:取食为害性及解毒酶活性变化初探

为了探究草地贪夜蛾对小麦的为害风险,我们以取食玉米作为对照,开展研究取食小麦对草地贪夜蛾的存活率、取食选择性、生长量和3种解毒酶及乙酰胆碱酯酶的影响。结果表明:草地贪夜蛾初孵幼虫在小麦上的存活率与玉米并无显著差异,高达90%以上。用小麦连续饲喂3龄幼虫7 d,日均体重增加迅速,与取食玉米无显著差异(P0.05)。2龄幼虫取食选择率为小麦玉米,3龄和4龄幼虫的取食选择率为玉米小麦,但均无显著差异。值得注意的是,小麦饲喂48 h后草地贪夜蛾谷胱甘肽S-转移酶活性和多功能氧化酶含量表现为显著诱导上升,而羧酸酯酶和乙酰胆碱酯酶的活性无显著变化。本试验结果初步表明草地贪夜蛾存在为害小麦的风险,应加强草地贪夜蛾在麦田发生情况的监测,同时应重视草地贪夜蛾取食不同寄主对药剂敏感性机制的研究。     

马铃薯晚疫病菌效应子PITG_16427.2的基因序列分析及功能验证

 马铃薯晚疫病的病原是致病疫霉菌(Phytophthora infestans),该病原菌在侵染马铃薯过程中分泌大量RxLR型效应子,但目前绝大多数RxLR效应子的功能和作用机制尚不明确。本研究成功克隆了致病疫霉菌的一个RxLR效应子PITG_16427.2,在本氏烟中瞬时表达PITG_16427.2,发现该效应子能够抑制6种激发子(INF1、PsojNIP、BAX、SIF2、Avh238、Avh241)激发的植物免疫反应。进一步研究发现,效应子PITG_16427.2在晚疫病菌侵染马铃薯早期上调表达。在15个致病疫霉菌株和3个同属菌株中克隆该基因,克隆到氨基酸序列一致性超过93%的同源基因,这些同源基因均能在本氏烟中抑制INF1和Avh241引起的HR,揭示了该效应子在病原卵菌中序列和功能的高度保守性。在本氏烟和马铃薯感病品种Désirée中瞬时表达PITG_16427.2,发现该效应子能够显著促进晚疫病菌的侵染。通过qRT-PCR方法检测发现,乙烯信号的相关基因ERF1显著上调,而水杨酸信号相关基因PR1b显著下调,表明PITG_16427.2在晚疫病菌侵染过程中可抑制寄主的SA信号途径,促进晚疫病菌侵染。因此,RxLR型效应子PITG_16427.2是致病疫霉菌中一个重要的侵染致病因子。     

农业生防昆虫黄猄蚁对3种农药的药剂敏感性研究

黄猄蚁(Oecophylla smaragdina)是最早用于农业生防的昆虫,目前已被证明能防治20多种农业害虫,对农林生产具有重要作用,但目前黄猄蚁对常见农药的药剂敏感性尚不清楚。本研究采集云南西双版纳地区的黄猄蚁,使用3种常见农药噻虫嗪、吡虫啉、呋虫胺对其进行了毒力测定,并在培养皿放置毒饵和蚂蚁之后的8 h、16 h、24 h、32 h、40 h、48 h分别记录了黄猄蚁的死亡情况。结果表明: 黄猄蚁对不同药剂的敏感性存在差异,黄猄蚁对呋虫胺,噻虫嗪以及吡虫啉的16h半数致死浓度(LC50)分别为0.130 g/l,0.290 g/l和0.382 g/l,差异显著。连续观察两天后,发现不同农药对黄猄蚁的致死时间也存在差异,该研究可为黄猄蚁生防的利用和保护提供科学参考。     

养殖大菱鲆肠道中大菱鲆弧菌的分离鉴定及药敏试验

2014年11月,大连地区某大菱鲆养殖场发生病害并伴有死亡,主要症状为吻部下颌明显出血,腹腔积水,肝脏充血,肠道出血有白便。从患病大菱鲆肠道中分离出1株可在硫代硫酸盐—柠檬酸盐—胆盐—蔗糖琼脂培养基上生长的菌株HZ-C1,人工回接感染试验证明其对健康大菱鲆具有较强致病性。通过形态学观察、16SrDNA序列同源性分析及生理生化试验确定菌株HZ-C1为大菱鲆弧菌。药敏检测结果显示,大菱鲆弧菌HZ-C1对氯霉素、丁胺卡那及头孢曲松等11种抗生素敏感,但对青霉素、四环素及诺氟沙星等15种抗生素耐药。本研究为大连地区养殖大菱鲆细菌性病害防治提供了一定病原学依据。     

Genetic diversity and antibiotic susceptibility of Staphylococcus aureus isolates from wild boars

We here report the occurrence of S. aureus in wild boars and characterize isolates genotypically and phenotypically in order to get knowledge about the occurrence of clonal lineages and genotypes in free-living wild animals. Forty-one S. aureus isolates obtained from 111 wild boars hunted in Lower Saxony, Germany, were investigated and compared to human and livestock isolates. The S. aureus belonged to multilocus sequence types ST1, ST7, ST30, ST133, ST425, ST804, ST890 and to the new ST3237, ST3238, ST3255 and ST3369. The livestock associated CC398-MRSA lineage, however, was not found. In addition to well-known spa types, the new types t14999, t15000, t15001 and t15002 were detected. Macrorestriction analysis revealed a variety of different SmaI fragment patterns. Most isolates were susceptible to all antimicrobials tested, including methicillin, and resistance was detected only to ampicillin, penicillin and erythromycin. PCR analysis confirmed the presence of staphylococcal enterotoxin genes (seh) in all t127-ST1 isolates. A high degree of genetic diversity was detected with many spa types and clonal lineages previously reported in humans and livestock animals.     

Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

As suggested by the Office International des Epizooties (OIE), fishes belonging to the genus Oplegnathus are more sensitive to megalocytivirus infection than other fish species including red sea bream (Pagrus major). To assess the roles of the innate immune response to these different susceptibilities, we cloned the genes encoding inflammatory factors including IL‐8 and COX‐2, and the antiviral factor like Mx from red sea bream for the first time and performed phylogenetic and structural analysis. Analysed expression levels of IL‐1β, IL‐8 and COX‐2 and the antiviral factor like Mx genes performed with in vivo challenge experiment showed no difference in inflammatory gene expression or respiratory burst activity between red sea bream and rock bream (Oplegnathus fasciatus). However, the Mx gene expression levels in red sea bream were markedly higher than those in rock bream, suggesting the importance of type I interferon (IFN)‐induced proteins, particularly Mx, during megalocytivirus infection, rather than inflammation‐related genes. The in vitro challenge experiments using embryonic primary cultures derived from both fish species showed no difference in cytopathic effects (CPE), viral replication profiles, and inflammatory and Mx gene expression pattern between the two fish species.     

Cyprinid herpesvirus 3 as a potential biological control agent for carp (Cyprinus carpio) in Australia: susceptibility of non‐target species

Carp (Cyprinus carpio L.) is a pest species in Australian waterways, and cyprinid herpesvirus 3 (CyHV‐3) is being considered as a potential biological control (biocontrol) agent. An important consideration for any such agent is its target specificity. In this study, the susceptibility to CyHV‐3 of a range of non‐target species (NTS) was tested. The NTS were as follows: 13 native Australian, and one introduced, fish species; a lamprey species; a crustacean; two native amphibian species (tadpole and mature stages); two native reptilian species; chickens; and laboratory mice. Animals were exposed to 100–1000 times the approximate minimum amount of CyHV‐3 required to cause disease in carp by intraperitoneal and/or bath challenge, and then examined clinically each day over the course of 28 days post‐challenge. There were no clinical signs, mortalities or histological evidence consistent with a viral infection in a wide taxonomic range of NTS. Furthermore, there was no molecular evidence of infection with CyHV‐3, and, in particular, all RT‐PCRs for viral mRNA were negative. As a consequence, the results encourage further investigation of CyHV‐3 as a potential biocontrol agent that is specific for carp.     

Isolation,Identification and Analysis of Serotype and Antibiotic Susceptibility of Pathogenic Salmonella from Chickens

To investigate the situations of predominant strain and antibiotic resistance of pathogenic Salmonella from chickens in Guangxi Zhuang Autonomous region, Salmonella was pre-enriched and isolated from tissues of clinical suspected chickens, and the Salmonella isolates were identified by biochemical test using ID 32E System for the identification of Enterobacteriaceae of VITEK System ATB Expression, serotypes were determined by slid agglutination test, antibiotic susceptibility was analyzed by ATB VET Susceptibility Test Strip of Enterobacteriaceae antibiotics.The results showed that 34 Salmonella strains were obtained from 310 clinical samples, of which 1 strain belonged to A serogroup, 1 to C2 serogroup, 15 to B serogroup, 14 to D serogroup and 3 to untyped serogroup, and S.typhimurium of B serogroup and S.pullorum of D serogroup were the predominant serotypes.All Salmonella isolates were sensitive to meropenem, imipenem, spectinomycine and apramycine, and the resistance rates to chloramphenicol, kanamycine and gentamicine were less than 10%, and the resistance rates to amoxicillin, streptomycine, flumequine, oxolinique, sulfamethizol, tetracycline and nitrofurantoine ranged between 50% and 90%, while the resistance rates to penicilline, oxacilline, fusidique, rifampicine and metronidazole reached to 100%.The results indicated that the serotype distribution of pathogenic Salmonella from chickens exhibited regional characteristics, and S.typhimurium and S.pullorum were the predominant serotypes in Guangxi Zhuang Autonomous region, and antibiotic resistance of Salmonella isolates was very serious which should be highly concerned.