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.     

Two-year surveillance of tilapia lake virus (TiLV) reveals its wide circulation in tilapia farms and hatcheries from multiple districts of Bangladesh

Tilapia lake virus (TiLV) is an emerging pathogen in aquaculture, reportedly affecting farmed tilapia in 16 countries across multiple continents. Following an early warning in 2017 that TiLV might be widespread, we executed a surveillance programme on tilapia grow-out farms and hatcheries from 10 districts of Bangladesh in 2017 and 2019. Among farms experiencing unusual mortality, eight out of 11 farms tested positive for TiLV in 2017, and two out of seven tested positive in 2019. Investigation of asymptomatic broodstock collected from 16 tilapia hatcheries revealed that six hatcheries tested positive for TiLV. Representative samples subjected to histopathology confirmed pathognomonic lesions of syncytial hepatitis. We recovered three complete genomes of TiLV from infected fish, one from 2017 and two from 2019. Phylogenetic analyses based on both the concatenated coding sequences of 10 segments and only segment 1 consistently revealed that Bangladeshi TiLV isolates formed a unique cluster within Thai clade, suggesting a close genetic relation. In summary, this study revealed the circulation of TiLV in 10 farms and six hatcheries located in eight districts of Bangladesh. We recommend continuing TiLV-targeted surveillance efforts to identify contaminated sources to minimize the countrywide spread and severity of TiLV infection.     

Comparison of effects of dietary‐specific fatty acids on growth and lipid metabolism in Nile tilapia

The dominant fatty acids (FAs) in oils are often used to explain different nutritional effects of dietary oils in fish. However, the amounts of dominant FAs among oils are different, and the nutritional roles of these important FAs in fish have not been precisely compared at similar levels in feeding trials. In the present study, different amounts of palmitic acid were added to safflower oil (SO), olive oil (OO) and fish oil (FO) to obtain comparable amounts (about 550 g/kg of total FAs) of 18:2n‐6, 18:1n‐9 and 20:5n‐3 + 22:6n‐3 and subsequently fed to Nile tilapia (11.1 ± 0.01 g) for 8 weeks. The results showed similar growth among groups but FO group obtained lower fat deposition, serum ALT and AST activities, compared to OO. Lipogenesis‐related gene expressions were higher in OO group than FO group in liver, muscle and adipose tissue, but there were only few differences in these genes between SO and FO groups. Lipid catabolism genes in FO group were higher than OO and SO groups in adipose tissue, but not in muscle, and the significantly higher expressions of CPT1b and PPARα were only observed in liver. Overall, dietary 18:2n‐6, 20:5n‐3 and 22:6n‐3 were beneficial to normal growth and lipid metabolism, whereas high amount of 18:1n‐9 induced lipid deposition and liver damage in Nile tilapia.     

5味中药醇提物对尼罗罗非鱼白细胞免疫活性的作用

将超微五倍子、白头翁、地榆、柯子、虎杖粉碎后过160目筛,按照料液比1∶10将药粉与60%的乙醇水溶液混合,再用超声波粉碎机超声1 h,用滤纸过滤收集滤液,置于多样品平行蒸发仪中浓缩后制得中药提取物。用吸取0.2 mL抗凝剂的无菌注射器自尼罗罗非鱼尾静脉采血0.5 mL,与L-15细胞培养基按体积比1∶9的比例混合成血浆,与中草药提取液单用、联用共孵育,利用氮蓝四唑还原法和流式细胞仪技术分别测定中草药提取液对尼罗罗非鱼外周血白细胞呼吸爆发活性和白细胞吞噬活性的影响。试验结果显示,五倍子、白头翁、地榆、柯子、虎杖单用均能显著提高白细胞氧呼吸爆发活性,呼吸爆发增加率分别为128.68%、123.96%、28.37%、44.44%和78.93%;试验组白细胞吞噬率分别较对照组增加44.04%、44.66%、95.00%、10.35%和64.93%。在两味中药双联用试验中,对白细胞氧呼吸爆发活性呈协同作用的组合为白头翁+地榆,呼吸爆发联用指数为1.28;对白细胞吞噬活性呈协同作用的组合为白头翁+虎杖、柯子+虎杖、五倍子+虎杖和白头翁+柯子,吞噬联用指数分别为11.45、6.20、3.03和1.06。     

Impact of sublethal di‐n‐butyl phthalate on the aquaculture fish species Nile tilapia (Oreochromis niloticus): histopathology and oxidative stress assessment

Phthalates such as di‐n‐butyl phthalate (DBP) and their esters are widely used plasticizers, their ubiquitous presence in daily life, inevitably leads to their restricted use due to important environmental pollution and health impacts and endocrine disruption potential. The aim of this study was to examine the effects of a sublethal concentration of 10 mg L^?1 DBP on haematocrit (HCT) values, gills and liver histology, malondialdehyde (MDA, 2‐thiobarbituric acid‐TBA reactivity) and reduced glutathione (GSH) levels in gills and liver tissue as oxidative stress biomarkers in the aquaculture fish species Nile tilapia (Oreochromis niloticus) after 24 (DBP‐24) and 96 (DBP‐96) h exposure. No differences were found between per cent HCT values in the 24 h exposure groups (P > 0.05). Response of antioxidant defence systems in liver and gill tissues of the fish were dependent on exposure duration and changed to a higher extent during 96 h. MDA levels in liver tissue increased in DBP treated fish in comparison to the control fish. However, the differences between the exposure and control groups were not significant (P > 0.05). A statistically significant decrease (P > 0.05) was recorded in gill MDA levels in the DBP‐96 group when compared to the control and DBP‐24 groups. The liver GSH levels were unchanged in the DBP treated fish. However, GSH levels were significantly lower in the gill tissue of the DBP‐96 group. Exposure to DBP caused several degenerative changes in the histology of gill and liver tissue. Gills displayed hyperaemia, epithelial lifting, oedema, talengiectasia, epithelial hyperplasia and fusion of secondary lamellae, whereas in liver several circulatory anomalies (hyperaemia, blood congestion and sinosoid dilatation) and vacuolization of hepatocytes were observed. Histopathological results demonstrated that the gills were more affected than the liver perhaps due to their direct contact with DBP.     

Single nucleotide polymorphisms in the growth hormone gene of Oreochromis niloticus and their association with growth performance

Polymorphisms in the growth hormone (GH) gene that is associated with the growth rate of farmed fish have been the target of many breeding programmes. The present study aimed to identify single nucleotide polymorphisms (SNPs) in GH gene regions to evaluate the association of SNP variations with the growth rate of two Nile tilapia: Oreochromis niloticus (Linnaeus, 1758) strains. The targeted regions were amplified, sequenced, aligned and screened for the presence of SNPs; thereafter, performance tests were used to check for the association between SNPs and weight. Allele and genotype frequencies were estimated for each SNP and genotype. Genotype blocks or sets of SNP genotypes and frequencies were also estimated. Association between SNPs and growth rate was statistically evaluated using a univariate linear mixed model that included both fixed and random effects. A total of 10 SNPs were identified, nine in the proximal promoter and one located in the 5′ UTR, forming 10 genotype blocks. In all weight recordings, five genotype blocks were significantly associated with the highest weights. Single nucleotide polymorphisms 6‐10 were also found to be significantly associated with growth (p‐value < .05). Genotypes with higher additive genetic values for weight were identified in the Chitralada strain, suggesting a possible impact of these additive effects of the GH SNP genotype on the growth rate of Nile tilapia. These findings may be used as part of marker‐assisted selection in tilapia breeding programmes.     

Semisynthetic ferulic acid derivative: an efficient feed additive for Genetically Improved Farmed Tilapia (Oreochromis niloticus)

Ferulic acid (FA) derivative was synthesized by modifying the functional groups of FA as dietary additive in the feeding of Genetically Improved Farmed Tilapia (GIFT) juveniles. Structural confirmation of this compound was based on IR, ¹H and ¹³C NMR techniques. This study was conducted to evaluate the effects of dietary FA derivative supplementation on growth performance, antioxidant status and serum biochemical parameters of GIFT juveniles compared with that of FA. A total of 675 fish with an average initial weight of 10.73 ± 0.40 g were fed nine diets supplemented with 0, 0.26, 0.52, 1.04 and 2.08 mmol kg^?1 FA, or 0.26, 0.52, 1.04 and 2.08 mmol kg^?1 FA derivative. Each diet was assigned to three replicate groups of 25 experimental tilapia for 56 days. Weight gain (WG) in groups of fish fed FA derivative was much higher than that of fish fed FA diet (P < 0.05). The antioxidant enzyme activities (SOD, CAT and GPx) in serum and liver of tilapia fed the FA derivative diets were significantly higher, while the malondialdehyde values were significantly lower than those of the fish fed the FA diets (P < 0.05). The serum biochemical parameters of GIFT were significantly improved by the FA derivative supplement. Moreover, feed conversion ratio, hepatosomatic index and viscerosomatic index were significantly decreased in fish fed FA or FA derivative diet at 0.52 mmol kg^?1 feed (P < 0.05). The investigation on the semisynthetic FA derivative provides a new way for developing safe and efficient feed additives.     

Effects of supplementary feeds with different protein levels on growth and economic performances of Nile tilapia (Oreochromis niloticus) cultured in a rain-fed rice–fish ecosystem

This study investigated the effects of supplementary feeds with different crude protein (CP) levels on the growth and economic performances of sex-reversed Nile tilapia (Oreochromis niloticus) reared in a rain-fed rice–fish ecosystem for a period of 120 days. There were four treatments—namely, traditional feed (21% CP), formulated feed (26% CP), commercial feed (30% CP), and control (no feed was used)—with three replicates per treatment. Fish (30.04 g) were stocked in ditches at a rate of 0.5 fish m^?2 considering the total area and fed one of the tested feeds twice daily at the rate of 2%–5% of body weight. Water-quality parameters of the ditches were monitored fortnightly, and these parameters were found suitable for fish farming. Organic matter, total nitrogen, and available phosphorous in soil were increased significantly (P < 0.05) in the feed treatments after fish cultivation. Highly significant weight gain and SGR were observed with fish fed the commercial feed as compared with other treatments. The highest benefit-cost ratio (2.15) was found in the control, followed by formulated feed (1.90), but in the case of numerical net return, it was the opposite. However, commercial feed showed the lowest benefit-cost ratio (1.67) due to higher feed cost than other supplementary feeds. Therefore, this study suggests that fish fed with a formulated feed (26% CP) produced significantly higher net economic return in the rice–fish farming system.     

Modeling and implementing the use of aeration to increase water temperature and dissolved oxygen in greenhouse aquaculture cages

In this study, an unique bamboo floating cage was built in order to heat the water used for rearing Nile tilapia (Oreochromis mossambicus x O. niloticus) and climbing perch (Anabas testudineus, Bloch) in Northern Thailand during the colder rainy and winter seasons. This special greenhouse fish cage (SGFC) was covered with polyethylene to heat the surface air with solar energy, and insulated to prevent heat loss. The water temperature at depths of 0.1, 0.3 and 0.8 m was modeled and measured. In various trials, the cages were aerated with and without heated air from the surface. It was found that the thermal efficiency of the SGFC integrated with hot air aeration at a water depth of 0.80 m was 32.65 %. Throughout the day, the average water temperature was 31.82 ± 0.68 °C, which was 0.76 °C higher than the water temperature outside the experiment, and the amount of oxygen in the water of 11.35 ± 0.13 mg/l. The SGFC can produce one extra fish harvest per year when compared with regular fish cages. Development equations to describe the thermal model were used for predicting the water temperature in the SGFC and thus validating the model. Moreover, the economic analysis of the SGFC found that the payback period was one year, seven months and fourteen days.     

Generation of mouse monoclonal antibodies specific to tilapia immunoglobulin using fish immunoglobulin/BSA complex for monitoring of the immune response in Nile tilapia Oreochromis niloticus

The simple immunoprecipitation method was used to isolate tilapia immunoglobulin (Ig) for immunization in order to produce monoclonal antibodies (MAbs) specific to tilapia Ig. First, the tilapia antiserum against bovine serum albumin (BSA) was prepared by peritoneal injection of BSA into tilapia, and the tilapia anti‐BSA antiserum was used to precipitate BSA to form the Ig/BSA immune complex. The Ig/BSA immune complex was then injected into Swiss mice for hybridoma production. After fusion, three hybridoma clones producing MAbs specific to the tilapia antibody were selected by dot blot and Western blot. All MAbs (101A, 59G, and 11A) were bound specifically to the heavy chain of immunoglobulin M (IgM). The MAbs 101A and 59G demonstrated twofold higher affinity than MAb 11A and the commercialized antibody. However, MAbs 11A could also bind to the heavy chain of IgM in Asian seabass, Lates calcarifer, as well. These MAbs can be used to monitor the immune responses of individual fish by indirect ELISA upon exposure to various antigens.