Tilapia Lake Virus was not detected in non-tilapine species within tilapia polyculture systems of Bangladesh

Sixteen countries, including Bangladesh, have reported the presence of tilapia lake virus (TiLV), an emerging tilapia pathogen. Fish polyculture is a common farming practice in Bangladesh. Some unusual mortalities reported in species co-cultivated with TiLV-infected tilapia led us to investigate whether any of the co-cultivated species would also test positive for TiLV and whether they were susceptible to TiLV infection under controlled laboratory experiments. Using 183 samples obtained from 15 farms in six districts across Bangladesh, we determined that 20% of the farms tested positive for TiLV in tilapia, while 15 co-cultivated fish species and seven other invertebrates (e.g. insects and crustaceans) considered potential carriers all tested negative. Of the six representative fish species experimentally infected with TiLV, only Nile tilapia showed the typical clinical signs of the disease, with 70% mortality within 12 days. By contrast, four carp species and one catfish species challenged with TiLV showed no signs of TiLV infection. Challenged tilapia were confirmed as TiLV-positive by RT-qPCR, while challenged carp and walking catfish all tested negative. Overall, our field and laboratory findings indicate that species used in polycultures are not susceptible to TiLV. Although current evidence suggests that TiLV is likely host-specific to tilapia, targeted surveillance for TiLV in other fish species in polyculture systems should continue, in order to prepare for a possible future scenario where TiLV mutates and/or adapts to new host(s).     

Blood and liver biopsy for the non‐destructive screening of tilapia lake virus

Detection of tilapia lake virus (TiLV) in tilapines is mainly from visceral organs of killed fish. However, lethal sampling might not be viable to broodstock and economically important ornamental cichlids. To contribute towards screening of the virus in asymptomatic infected fish, a subclinically infected population of Nile tilapia adults obtained from a local farm was preliminarily tested to compare different non‐lethal sampling methods, for example liver biopsy, gill biopsy, fin clip, mucus, faeces and blood for detection of TiLV. Only liver and blood samples gave positive results by PCR. Since blood sampling is relatively simpler, it was further used for five naturally co‐cultured juvenile fish species from above‐mentioned farm including 40 red tilapia broodstock and 20 Nile tilapia adults from two other different farms. The results showed that from the tested fish, 4 of 5 Nile tilapia, 2 of 5 hybrid red tilapia and 3 of 5 giant gourami blood samples tested positive, while 38 of 40 blood samples of red tilapia tested positive for TiLV in second‐step PCR. Sequencing representative PCR amplicons of positive samples confirmed sequence identity to TiLV. In conclusion, both blood and liver biopsy are practical non‐destructive sampling platforms for TiLV screening in cichlids with blood being more convenient, especially for tilapia broodstock.     

Evidence of potential vertical transmission of tilapia lake virus

Tilapia lake virus disease (TiLVD) is an emerging viral disease in tilapia with worldwide distribution. Although the horizontal transmission of TiLV has been demonstrated through the cohabitation of infected fish with susceptible fish, no direct experiment showed the potential of vertical transmission from broodstock to progeny. In this study, natural outbreaks of TiLV in broodstock and fry in two tilapia hatcheries were confirmed. The TiLV genomic RNA was detected in liver and reproductive organs of infected broodstock, while infective virus was isolated in susceptible cell line. In situ hybridization assay confirmed the presence of TiLV in the ovary and testis of naturally infected fish and experimentally challenged fish. Moreover, early detection of TiLV in 2‐day‐old fry and the presence of TiLV genomic RNA and viable virus in the testis and ovary suggested the possible transfer of this virus from infected broodstock to progenies. As infective virus was present in gonads and fry in natural outbreak and experimental fish, the importance of biosecurity and prevention of the virus to establish in the hatchery should be emphasized. Hence, the development of TiLV‐free broodstock and the maintenance of high biosecurity standards in the hatcheries are essential for any attempt of virus eradication.     

Assessing the population transmission dynamics of tilapia lake virus in farmed tilapia

A novel virus, tilapia lake virus (TiLV), has been identified as a key pathogen responsible for disease outbreak and mass mortality of farmed tilapia. We used a deterministic susceptible‐infectious‐mortality (SIM) model to derive key disease information appraised with published TiLV‐induced cumulative mortality data. The relationship between tilapia mortality and TiLV exposure dosages was described by the Hill model. Furthermore, a disease control model was proposed to determine the status of controlled TiLV infection using a parsimonious control reproduction number (R_C)‐control line criterion. Results showed that the key disease determinants of transmission rate and basic reproduction number (R₀) could be derived. The median R₀ estimate was 2.59 in a cohabitation setting with 2.6 × 10⁵ TCID50 fish^?1 TiLV. The present R_C‐control model can be employed to determine whether TiLV containment is feasible in an outbreak farm by quantifying the current level of transmission. The SIM model can then be applied to predict what additional control is required to manage R_C < 1. We offer valuable tools for aquaculture engineers and public health scientists the mechanistic‐based assessment that allows a more rigorous evaluation of different control strategies to reduce waterborne diseases in aquaculture farming systems.     

Tilapia lake virus: The story so far

Tilapia lake virus (TiLV) is a highly contagious pathogen that has detrimental effects on tilapia farming. This virus was discovered in 2014 and has received tremendous global attention from the aquaculture sector due to its association with high fish mortalities and its strong economic impact on the tilapia aquaculture industry. Currently, TiLV has been reported in 16 countries, and this number is continuing to rise due to improved diagnostic assays and surveillance activities around the world. In this review, we summarize the up-to-date knowledge of TiLV with regard to TiLV host species, the clinical signs of a TiLV infection, the affected tissues, pathogenesis and potential disease risk factors. We also describe the reported information concerning the virus itself: its morphology, genetic make-up and transmission pathways. We review the current methods for virus detection and potential control measures. We close the review of the TiLV story so far, by offering a commentary on the major TiLV research gaps, why these are delaying future TiLV research and why the TiLV field needs to come together and proceed as a more collaborative scientific community if there is any hope limiting the impact of this serious virus.     

Assessment of the impact of dissemination of genetically improved Abbassa Nile tilapia strain (GIANT‐G9) versus commercial strains in some Egyptian governorates

WorldFish initiated a selective breeding program in Abbassa—Egypt to develop and produce the genetically improved Nile tilapia strain known as “Genetically Improved Abbassa Nile tilapia (GIANT)”, adopting the same technology used for the Genetically Improved Farmed Tilapia (GIFT), in Asia. WorldFish provided the Ninth Generation (G9) broodstock of the GIANT to 11 Broodstock Multiplication Centers (BMC’s) in five governorates; these centres then disseminated improved mixed‐sex fry to 160 tilapia hatcheries which supplied all‐male fry to 1,500 fish farms in 2017. We present results of an impact assessment of the G9 of GIANT in 83 of these fish farms in four Egyptian governorates (Kafr El‐Sheikh, Fayoum, Behera and Alexandria). Results indicate that the use of GIANT in all governorates achieved significantly higher (18.8%) fish yields (12.3%–26.4%) and 15.7% lower FCR (feed conversion ratio), reflecting a positive impact on their net profits compared with those who used commercial strains. Overall, fish farmers who stocked GIANT had significantly higher average fish sales (5,567 US$/ha) than those who stocked non‐improved commercial strains (5,192 US$/ha) in all governorates. However, when comparing within governorates, the difference was significant only in Fayoum and Alexandria noting a mixed performance of the GIANT strain. The mixed performance indicates that some hatcheries may have crossed the different strains, which lowered the performance differences. To get an objective analysis of the performance of the GIANT, it is recommended that similar assessment be undertaken with new generations of the strain supported by on‐farm performance trials.     

Detection of tilapia lake virus (TiLV) infection by PCR in farmed and wild Nile tilapia (Oreochromis niloticus) from Lake Victoria

Tilapia lake virus disease (TiLVD) has emerged to be an important viral disease of farmed Nile tilapia (Oreochromis niloticus) having the potential to impede expansion of aquaculture production. There is a need for rapid diagnostic tools to identify infected fish to limit the spread in individual farms. We report the first detection of TiLV infection by PCR in farmed and wild Nile tilapia from Lake Victoria. There was no difference in prevalence between farmed and wild fish samples (p = .65), and of the 442 samples examined from 191 fish, 28 were positive for TiLV by PCR. In terms of tissue distribution, the head kidney (7.69%, N = 65) and spleen (10.99%, N = 191), samples had the highest prevalence (p < .0028) followed by heart samples (3.45%, N = 29). Conversely, the prevalence was low in the liver (0.71%, N = 140) and absent in brain samples (0.0%, N = 17), which have previously been shown to be target organs during acute infections. Phylogenetic analysis showed homology between our sequences and those from recent outbreaks in Israel and Thailand. Given that these findings were based on nucleic acid detection by PCR, future studies should seek to isolate the virus from fish in Lake Victoria and show its ability to cause disease and virulence in susceptible fish.     

Virucidal effects of common disinfectants against tilapia lake virus

Tilapia lake virus (TiLV) is an emerging virus associated with high fish mortality and economic losses. This study investigates the virucidal effects of the following disinfectants (active ingredients) on TiLV: 2.5 ppm iodine, 10 ppm sodium hypochlorite (NaOCl), 300 ppm hydrogen peroxide (H₂O₂), 80 ppm formalin and 5,000 ppm (0.5%) Virkon^®. Factors that affect the disinfectants’ efficacy, including temperature, contact time and soiling (organic matter) interference, were examined under conditions mimicking natural aquaculture practices. TiLV inactivation of higher than 5 log₁₀ TCID₅₀ ml^?1 was achieved after 10 min and at 28°C for all disinfectants except formalin; similar inactivation levels were reached by NaOCl and Virkon^® at 10 min and 4°C. Extended exposure to formalin from 10 to 60 min at 28°C rendered more than 5 log₁₀ inactivation. Increasing synthetic organic matter in the water to mimic soiling interference reduced the efficacy of NaOCl, iodine and H₂O₂ when tested at 10 min and 28°C; however, Virkon^® still achieved more than 5 log₁₀ inactivation. This study demonstrates that most common disinfectants effectively reduced viral loads to minimum levels. To limit the spread of TiLV in aquaculture farms and related facilities, the appropriate use of such disinfectants should therefore be promoted and implemented.     

Economic efficiency of small-scale tilapia farms in Guangxi,China

ABSTRACT

The economic efficiency of small-scale tilapia farms in Guangxi, China were analyzed by using Data Envelopment Analysis (DEA). The mean technical (under CRS and VRS), allocative, and economic efficiencies of small-scale tilapia farms were 0.89, 0.97, 0.71 and 0.68, respectively. Efficiency analysis revealed that inefficient farms would have needed to increase production output levels by 32% to perform as well as the best practice farms. The decomposition of the economic efficiency showed that allocative inefficiency was the primary cause of economic inefficiency. There were positive relationships between economic efficiency and age, culture mode and period through Tobit regression, while experience, family members, and technology support had negative effects on economic efficiency. Lower input levels of feed quantity may help to increase the tilapia farming economic efficiency.     

罗非鱼湖病毒核蛋白的克隆表达、抗体制备及其组织分布

近年来罗非鱼湖病毒(TiLV)在多个国家流行,对世界罗非鱼养殖业造成严重威胁。中国是罗非鱼第一养殖大国,尽管我国大陆还没有TiLV的正式报道,鉴于吉富罗非鱼是我国重要的罗非鱼养殖品种,其对TiLV的感染特性研究具有重要意义。本实验采用TiLV对吉富罗非鱼进行人工感染,随后在肝脏组织中克隆和测定了TiLV第6片段基因。罗非鱼湖病毒第6片段基因cDNA全长1044 bp,开放读码框(ORF)为954 bp,编码317个氨基酸,预测分子量为36.38 ku;5′非编码区(NCR)为19 bp,3′非编码区(NCR)为972 bp。系统进化树分析表明该蛋白属于TiLV核蛋白(NP)。随后在大肠杆菌中表达和提纯了GST融合NP蛋白,在新西兰大白兔上制备了多克隆抗体。通过酶联免疫吸附实验(ELISA)检测抗体效价为1∶51200,且抗体可特异性识别感染组织中的病毒NP蛋白。对吉富罗非鱼不同组织进行苏木精—伊红(H.E)染色观察,发现肝脏组织坏死并形成合胞体,脾脏部分细胞出现空泡、坏死,含铁血黄素增多,头肾细胞坏死,鳃丝上皮细胞明显解离脱落,鳃小片黏连,脑组织细胞肿大。通过蛋白印迹法(WB)和免疫组化(IHC)对人工感染TiLV的吉富罗非鱼不同组织进行检测,结果显示,NP蛋白在肝脏、脑、体肾和头肾等组织中均有表达,以肝脏组织中表达量最高。为了解吉富罗非鱼对TiLV的免疫反应,通过实时荧光定量PCR(qRT-PCR)测定免疫因子TNF-α和TGF-β在主要免疫器官脾脏和头肾组织中的表达。研究表明,在感染早期(感染后12~24 h),病毒可显著抑制TNF-α和TGF-β在脾脏和头肾中的表达,可能通过抑制宿主这些免疫因子来促进病毒自身早期的复制。本研究将为进一步解读TiLV的致病机理及其高效防控提供理论基础。     

The economics of tilapia fingerling production and marketing in the Philippines*

Abstract. These analyses of the tilapia fingerling industry in the Philippines focus on the economics of production in different hatchery systems and on fingerling marketing channels. The analyses are based on cross-sectional data from government and private hatcheries in late 1982 and from December 1983 to March 1984. The relative profitabilities of different hatchery systems (land-based and lake-based) are estimated and economies of scale are analysed. Conceptual and methodological issues inherent in analyses based on cross-sectional data are discussed in the context of the extreme difficult standardizing inputs and output from private tilapia hatcheries which are characterized by wide variation in management practices.     

罗非鱼罗湖病毒ORF10蛋白的亚细胞定位及组织表达分析

为了研究罗非鱼罗湖病毒(tilapia lake virus, TiLV) ORF10 蛋白的功能, 从染病莫桑比克罗非鱼(Oreochromis niloticus)脾脏和肝脏组织中克隆获得 TiLV ORF10 基因编码序列, 并成功构建了荧光定位载体 pEGFP-ORF10 和重组表达载体 pET32a-ORF10。将荧光定位载体 pEGFP-ORF10 转染鲤(Cyprinus carpio)上皮瘤细胞(Epithelioma papulosum cyprinid, EPC), 经荧光显微镜观察, 在 EPC 的细胞核中观察到绿色荧光信号, 表明该蛋白定位于细胞核中。将重组载体 pET32a-ORF10 转化大肠杆菌 BL21(DE3)进行目的融合蛋白表达, 聚丙烯酰胺凝胶电泳(SDS-PAGE)检测表明, His-TiLV ORF10 融合蛋白成功表达并主要存在于上清液中。随后, 采用 Ni-NTA 亲和层析的方法纯化融合蛋白 His-TiLV ORF10, 以其为抗原多次免疫 Balb/C 小鼠, 制备多克隆抗体。蛋白印迹法(WB)分析显示, 制备的抗体可以特异性识别 His-TiLV ORF10。利用蛋白印迹法进一步对感染 TiLV 莫桑比克罗非鱼的不同来源组织进行检测, 结果显示, TiLV ORF10 蛋白在染病莫桑比克罗非鱼肌肉、脾脏、肝脏和肾脏中的表达存在较大差异, 其中以肝脏和肾脏组织表达量为最高, 脾脏次之, 肌肉中表达量最低。本研究为深入了解 TiLV ORF10 蛋白的功能和病毒侵染与致病等机理提供了重要基础。     

Application of multiplex quantitative polymerase chain reaction methods to detect common bacterial fish pathogens in Nile tilapia,Oreochromis niloticus,hatcheries in Costa Rica

Edwardsiella spp., Streptococcus spp., and Francisella noatunensis subsp. orientalis are some of the most important fish pathogens affecting global tilapia, Oreochromis spp., aquaculture. In Costa Rica, the aquaculture industry is dominated by freshwater‐cultured Nile tilapia, Oreochromis niloticus, which are raised in all seven national provinces. At present, little is known regarding the diversity of pathogens present in these facilities, and definitive identification of agents associated with disease outbreaks are rare. To evaluate the prevalence of common bacterial pathogens in these systems, this study used multiplex quantitative polymerase chain reaction (qPCR) assays targeting Edwardsiella, Streptococcus, and Francisella species as a diagnostic and surveillance tool. In 2017, seven different tilapia hatcheries were visited, and 350 fingerlings were subjected to necropsy and molecular diagnostic evaluation. Fish exhibiting gross signs of disease were subjected to histological and microbiological analysis. For the first time, Edwardsiella anguillarum was recovered and molecularly confirmed from diseased tilapia in Costa Rica. In addition, F. noatunensis subsp. orientalis was identified in a region of Costa Rica where it had not been previously reported.     

Infectious pancreatic necrosis virus isolated from farmed rainbow trout and tilapia in Kenya is identical to European isolates

Infectious pancreatic necrosis virus (IPNV) is an aquabirnavirus that causes serious diseases in a variety of fish species worldwide. It has been isolated from a large number of healthy fresh and marine water fish. Prior to this study, there was no record of the presence of IPNV infection in Kenya. Here, the presence of IPNV in farmed rainbow trout and tilapia was examined in Nyeri County of central Kenya. Head kidney samples taken from five rainbow trout and three tilapia farms and stored in RNALater^® were processed by PCR followed by sequencing of a segment A fragment covering nucleotide positions 2,120–2,343 bp. IPNV was detected in all the farms sampled with infection ratios ranging from 0.3 to 0.78 although the infections were not associated with any specific clinical signs of disease. These findings were supported by immunohistochemistry staining of the virus in the kidney and exocrine pancreas of rainbow trout. Sequence alignment and phylogenetic analysis revealed that the Kenyan isolates were identical to European isolates, suggesting a common origin. These findings highlight the need for better biosecurity procedures with more stringent surveillance programmes and control for fish diseases, especially focusing on imported breeding materials to Kenya.     

Multi-criteria evaluation approach to GIS-based land-suitability classification for tilapia farming in Bangladesh

Site selection is a key factor in any aquaculture operation, because it affects both success and sustainability. It can, moreover, solve conflicts between different activities, making rational use of the land. This study was conducted to identify suitable sites for development of Nile tilapia (Oreochromis niloticus) farming in Sitakunda Upazila (sub-district), Bangladesh, using GIS-based multi-criteria evaluation of water and soil quality, topography, infrastructure and socio-economic factors. ASTER image and eighteen thematic layers were analyzed using ENVI and ArcView software to identify the suitable areas for tilapia farm development. A constraint layer was used to exclude areas from suitability maps that cannot be allowed to implement tilapia farming. A series of GIS models were developed to identify and prioritize the most suitable areas for tilapia farming. The output of the model clearly indicates the location and extent of tilapia farming areas on different suitability scales, i.e. most suitable (7,744 ha), moderately suitable (2,479 ha), and not suitable (838 ha). Model outputs were assessed against field verification data, and were consistent. Because existing aquaculture covers only 1,540 ha of land in the study area, the potential for expanding tilapia farms should take into consideration socio-political and environmental issues. The results are encouraging in terms of tilapia culture development and suggest that grassland–agriculture areas could be used for sustainable development of tilapia farming to diversify the economic activities of rural communities.