Experimental early pathogenesis of Streptococcus agalactiae infection in red tilapia Oreochromis spp.

Streptococcus agalactiae causes a severe systemic disease in fish, and the routes of entry are still ill‐defined. To address this issue, two groups of 33 red tilapia Oreochromis spp. each of 10 g were orally infected with S. agalactiae (n = 30), and by immersion (n = 30), six individuals were control‐uninfected fish. Three tilapias were killed at each time point from 30 min to 96 h post‐inoculation (pi); controls were killed at 96 h. Samples from most tissues were examined by haematoxylin–eosin (H&E), indirect immunoperoxidase (IPI) and periodic acid‐Schiff; only intestine from fish infected by gavage was evaluated by transmission electron microscopy. The results of both experiments suggest that the main entry site of S. agalactiae in tilapia is the gastrointestinal epithelium; mucus seems to play an important defensive role, and environmental conditions may be an important predisposing factor for the infection.     

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.     

Evaluation of the link between gyrodactylosis and streptococcosis of Nile tilapia, Oreochromis niloticus (L.)

Streptococcus iniae and Gyrodactylus niloticus are two common pathogens of cultured Nile tilapia, Oreochromis niloticus. We studied concurrent infection of tilapia by G. niloticus and S. iniae and evaluated whether parasitism in tilapia with Gyrodactylus increased susceptibility and mortality following immersion infection with S. iniae. Results showed that death mainly occurred in fish with G. niloticus and challenged with S. iniae (G-S group). The accumulative mortality (42.2%) was significantly higher in the G-S group than in fish not infected by the parasite (6.7%), but exposed to S. iniae. Bacteriological examination revealed S. iniae from > or =92% of dead or moribund fish challenged with S. iniae. Gyrodactylus not only damaged fish epithelium and provided entry for invasive bacteria but also was found to harbour viable cells of S. iniae for 24 and 72 h. Streptococcus iniae was isolated from 60% and 40% of G. niloticus collected from fish infected by intraperitoneal injection or immersion, respectively, at 24 h post-challenge. The present study confirms that parasitism of tilapia by G. niloticus increased host mortality following exposure to the bacterial pathogen S. iniae.     

Evaluation of rubber seed oil as lipid source in red tilapia (Oreochromis sp.) diet

Rubber seed oil has a high potency to be used as a new lipid source within red tilapia diet. However, the rubber seed oil contains antinutritional factor such as hydrogen cyanide (HCN). HCN is classified into heat‐labile; therefore, heating effectively reduces the HCN content. Based on this fact, this research evaluated the use of rubber seed oil with and without heating as lipid source in red tilapia diet. Four isonitrogenous and isocaloric diets were prepared as experimental diet. Each test diet contains different lipid sources such as crude palm oil + corn oil (CTRL), crude palm oil + rubber seed oil with heating (CPO:PRSO), 100% rubber seed oil with heating (PRSO) and 100% RSO without heating (URSO). In average, there were no significant differences (Tukey: p < .05) among test diet except diet with 100% rubber seed oil without heating. It concluded that 100% rubber seed with heating within test diet has no effect on growth, feed performance, blood profile, cholesterol plasma, body and liver glycogen, malondialdehyde (MDA) and the activity of superoxide dismutase (SOD).     

Molecular and histopathological characterization of Photobacterium damselae in naturally and experimentally infected Nile tilapia (Oreochromis niloticus)

Mass mortality has occurred among cultured Nile tilapia, Oreochromis niloticus, on fish farms in Manzala, Dakahlia province, Egypt, in the summer season, 2019. Moribund fish were reported with deep ulcers, septicaemic lesions and sampled for bacterial isolation. In this study, most isolates were subjected to bacteriological examination, antibiotic sensitivity test, 16S rRNA gene sequencing and histopathological examination. Following isolate identification, intraperitoneal challenge of Nile tilapia with a bacterial suspension 2 × 10⁶ CFU/ml was performed. Samples from liver, spleen and kidney were collected for histological and biochemical analysis. The results showed a high similarity (99%) to Photobacterium damselae strains using phylogenetic analysis of 16S rRNA. P. damselae exhibited resistance to amoxicillin and erythromycin, as well it was highly sensitive to chloramphenicol and doxycycline. Moreover, haemorrhage, oedema, hemosiderosis and melanomacrophage activation in the liver and head kidney of infected fish were detected by light and electron microscopy. Also, significant higher levels of CAT and SOD in the spleen and head kidney, as well as the serum levels of NO were observed in experimentally challenged O. niloticus, compared to the control fish. Our data identified P. damselae for the first time from infected Nile tilapia, describing its sensitivity to a variety of antibiotics, histopathological alterations and oxidative stress impact, and it could be useful indicators for understanding P. damselae pathogenesis, which might provide a preventive efficacy for P. damselae.     

Effect of temperature and phytoplankton concentration on Nile tilapia Oreochromis niloticus (L.) filtration rate

Nile tilapia Oreochromis niloticus (L.) held in timed‐pulse feeding chambers, were provided with algal‐rich water dominated by either green algae (Scenedesmus, Ankistrodesmus, Chlorella and Tetraedron) or cyanobacteria (Microcystis) to determine the effect of temperature and phytoplankton concentration on filtration rates. Green algae and cyanobacteria filtration rates were measured as suspended particulate organic carbon (POC) kg^?1 wet fish weight h^?1. Ivlev's filter‐feeding model described the relationships between filtration rates and suspended POC concentration of green algae and cyanobacteria. Filtration rates of both green algae and cyanobacteria increased linearly as water temperature increased from 17 °C to 32 °C and were significantly higher in the warm‐water regime (26–32 °C) than in the cool‐water regime (17–23 °C). Filtration rates at 95% saturation POC (FR₉₅) in green algal and cyanobacterial waters were 700 mg C kg^?1 h^?1 and 851 mg C kg^?1 h^?1 in the warm‐water regime and 369 mg C kg^?1 h^?1 and 439 mg C kg^?1 h^?1 in the cool‐water regime respectively. The FR₉₅ in warm water were achieved at lower POC concentrations than in cool water.     

Characterization of microbial communities in intensive GIFT tilapia (Oreochromis niloticus) pond systems during the peak period of breeding

Aquaculture pond is a complex ecosystem where the microorganisms in the sediments, in the animal intestinal tract and in water interact with each other to influence the water quality and health of aquatic animals. In order to understand the spatial distribution and relationship of microbial communities in intensively farmed genetically improved farmed tilapia (GIFT, Oreochromis niloticus), 454 high‐throughput pyrosequencing was applied to analyse the 16S rRNA gene of bacteria in intensive GIFT ponds in Wuxi City, Jiangsu Province, China. A total of 72 747 initial sequences were obtained from four depths of pond water, from tilapia large intestines and from pond sediment. The most common phylum in all samples was Proteobacteria, Actinobacteria and Cyanobacteria were the most abundant in water, Fusobacteria and Firmicutes in the large intestine and Chloroflexi in sediment samples. The sediment microbial community structure was comparatively similar to that of the tilapia large intestine. The microbial communities from different water depths were somewhat similar, especially the three most shallow samples, although the abundance of Actinobacteria gradually decreased with increased water depth. This data offer a preliminary exploration of the response mechanisms of the bacterial communities to aquafarming and contributes to the understanding of the status of bacterial communities of tilapia pond systems during the peak period of breeding from the aspect of their spatial distributions.     

Evaluation of antimicrobial products used in tilapia (Oreochromis spp.) and whiteleg shrimp (Litopenaeus vannamei) aquaculture

Despite the importance of aquaculture increases for feeding the growing world population, professional farm management and proper use of aquaculture products or drugs are needed. It is important to know the quality of commercial antimicrobial products on the market while there is wide use of antimicrobials to treat fish diseases. The most commonly used antimicrobial products were identified from a previous survey, included amoxicillin, enrofloxacin, florfenicol, sulphadiazine, sulphamonomethoxine and sulphadimidine. Eight local chemical supply shops were randomly sampled in representative aquaculture areas in Maoming and Zhanjiang districts of south China in 2013. In this study, only eight products contained active substances within ±10% of the concentration declared on the product label among the 30 different most used commercial products; while six products did not contain any of the declared antimicrobials. Three products without labels were sold as ‘pure antimicrobial’ and had high concentrations of active antimicrobials. Furthermore, most products only provided ambiguous information on the packages on both antimicrobial usage and dosage; and only one product stated how to prepare medicated feed which is important for efficient usage. The results of this study showed that the aquaculture antimicrobial products sold on the market were of low quality and thus compromised effective fish disease treatment. Moreover, insufficient information on products’ labels would provide limited instruction on correct antimicrobial usage. Inspection and supervision by both private companies and the government's sector require strengthening.     

Economic profitability of Nile tilapia (Oreochromis niloticus L.) production in Kenya

Economic profitability of Nile tilapia production in Kenya was analysed using a model that simulated individual fish growth and took fish population dynamics in the pond into account. The results suggest that the currently practiced mixed‐sex tilapia culture is economically unsustainable. It is suggested that research and extension efforts be geared towards developing monosex Nile tilapia production systems. Nile tilapia culture with African catfish predation should be viewed as an intermediate step towards all‐male Nile tilapia culture. This will allow accumulation of both physical and human capital to support all‐male tilapia culture. Under all‐male culture, economic returns are high enough to justify investment in Nile tilapia culture using borrowed capital. However, the success of monosex culture will depend on the availability and affordability of quality fingerlings and low‐cost fish feeds. The results have a wide application in Sub‐Saharan Africa where mixed‐sex Nile tilapia culture is common.     

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.     

Phenotypic and genotypic characterization of Streptococcus spp. isolated from tilapia (Oreochromis spp.) cultured in river-based cage and earthen ponds in Northern Thailand

Streptococcus spp. are major pathogenic bacteria associated with massive mortality in tilapia. This study investigated the phenotypic and genotypic characterization of Streptococcus agalactiae (GBS) and Streptococcus iniae (S. iniae) isolated from tilapia in river-based floating cage and earthen pond farms in northern Thailand. Isolates were identified by biochemical and molecular analyses. Capsular typing, enterobacterial repetitive intergenic consensus polymerase chain reaction and multilocus sequence typing were performed to investigate the genetic relatedness. Six and one isolates were confirmed as GBS and S. iniae, respectively. All Streptococcus spp. isolates were obtained from 4 river-based cage farms (4/33), while samples collected from earthen pond farms (N = 28) were negative for streptococcosis. All GBS with serotype Ⅲ and sequence type (ST) 283 was observed. The β-haemolytic GBS isolates were resistant to five antimicrobials, while the S. iniae was susceptible to all antimicrobials. This study indicates both GBS and S. iniae are the major bacterial pathogens responsible for streptococcosis infection in farmed tilapia of northern Thailand with GBS as dominant species. This survey highlights that the river-based cage farms seriously impact on the healthy development of the tilapia industry.     

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.     

Experimental evaluation of mass selection to improve red body colour in Fijian hybrid tilapia (Oreochromis niloticus × Oreochromis mossambicus)

Mass selection was trialed on Fijian hybrid tilapia (Oreochromis niloticus × Oreochromis mossambicus) to reduce the amount of black spots on otherwise red phenotypes. The responses of two selection intensities (low‐selection line, top 50% – L) and (high‐selection line, top 30% – H) were compared with that of a control line (no selection – C) across three generations. The relative growth performance of treatments (C, L and H) was examined in parallel in each generation to assess whether mass selection had a correlated negative effect on growth performance. The results show clearly that red phenotype can be improved significantly by applying mass selection, without affecting growth performance. We propose that black spots on an otherwise red phenotype could represent the allelic products of a second genetic locus influencing skin colour, which can be expressed in red individuals (genotype Rr) but which may be masked in black individuals (genotype rr).     

Growth response and acquired resistance of Nile tilapia, Oreochromis niloticus (L.) that survived Streptococcus iniae infection

This study determined the growth performance and acquired resistance of Nile tilapia, Oreochromis niloticus (L.) that survived Streptococcus iniae infection. Tilapia were challenged with three doses of S. iniae (8.8 × 10³, 8.8 × 10⁴ and 8.8 × 10⁵ CFU fish^?1 for low, medium and high challenges respectively). Groups of non‐injected and tryptic soy broth‐injected fish were maintained as controls. Significantly (P<0.05) higher mortality (45.0%) occurred in the high challenge treatment than in the low challenge treatment group (29.6%). The medium challenge group had mortality (36.3%) that did not differ significantly from the high or low treatment. Few fish died in the non‐injected and broth‐injected treatments (3.4% and 0.8% respectively). The tilapia that survived S. iniae infection used to assess growth performance were selected from survivors without gross clinical signs of disease. These fish were randomly stocked at a rate of 30 fish into each 57 L aquarium in triplicate and fed to apparent satiation for 8 weeks. No significant differences were detected in weight gain, feed intake, feed efficiency ratio or survival between S. iniae‐survived tilapia and the control treatments following the 8‐week growth performance trial. Following the 8‐week feeding study, tilapia were challenged with 1 × 10⁶ CFU fish^?1 of S. iniae to assess acquired immunity. Mean cumulative mortality was significantly higher (P<0.05) in the control treatments (41.7% for the non‐injected and 43.3% for the broth‐injected fish) than in the low, medium and high challenge treatments (7.4%, 3.3% and 8.3% respectively). Serum protein was significantly (P<0.05) elevated in the S. iniae‐survived tilapia that were subsequently challenged when compared with controls challenged for the first time. Agglutinating antibody titre was significantly higher in the fish in the medium and high challenge treatments, compared with the control fish challenged for the first time. The results suggest tilapia that survive S. iniae challenge without showing overt disease signs performed as well as non‐infected tilapia. Further, the S. iniae‐survived tilapia challenged following the 8‐week growth performance trial gained acquired resistance to homologous S. iniae challenge.     

Nutrient composition and sensory evaluation of fillets from wild-type Nile tilapia (Oreochromis niloticus, Linnaeus) and a red hybrid (Florida red tilapia × red O. niloticus)

A comparison of composition and sensory evaluation were performed on fillets of Oreochromis niloticus (wild type) and red hybrid tilapia (RHT) (Florida red tilapia ♂× red O. niloticus♀). Fifty male tilapia, 25 O. niloticus (initial weight 159.3 g) and 25 red hybrid (initial weight 132.4 g), were placed in each of three 2.0 m³ volume tanks. The fish were fed a commercial feed containing 35.9% crude protein during the 120‐day experimental period and then killed in cold water (3°C). All the fish were filleted. Twelve fillets from each treatment were used for proximate analyses, five fillets from each treatment were used for fatty acids analyses and the remaining fillets were used for sensory evaluation. The compositional analyses showed similar moisture, true protein and ash content in both genetic groups, but a lower crude fat content was measured in the red hybrid fillets (0.70%) compared with that of O. niloticus fillets (0.97%). No differences between O. niloticus and the red hybrid were observed in their fillet profile of fatty acids. In the sensory evaluation test, a difference in fillet flavour between both genetic groups was perceived. Of the 112 consumers in the preference test, 81.2% perceived a difference in fillet flavour between the two tilapias, with a general preference for the red hybrid over the wild‐type O. niloticus. The benefits of cultivating a RHT with a low lipid content are discussed.     

Francisella sp., an emerging pathogen of tilapia, Oreochromis niloticus (L.), in Costa Rica

Francisella sp. is an emergent bacterial pathogen that causes acute to chronic disease in warm and cold water cultured and wild fish species. During the past 3 years, the bacterium has been detected in tilapia, Oreochromis niloticus , cultured in Costa Rica. Infected fish presented non-specific clinical signs, such as erratic swimming, anorexia, anaemia, exophthalmia and high mortality. Upon macroscopic and microscopic examination, several internal organs (mainly spleen and kidney) were enlarged and contained white nodules. Histological examination revealed the presence of multifocal granulomatous lesions, with the presence of numerous small, pleomorphic, cocco-bacilli. The bacteria were isolated from infected tilapia on selective media and grown on several media with and without antibiotics. Specific PCR primers to the Francisella genus were used to confirm the preliminary diagnoses. In comparison with several bacterial 16S rRNA sequences, our isolate was found to share 99% identity with other Fransicella spp. isolated from fish, and more than 97% identity to the human pathogen Francisella tularensis . Koch's postulates were fulfilled after experimental intraperitoneal and gill exposure challenges.