Production of red tilapia (Oreochromis spp.) in floating cages in the Mekong Delta, Vietnam: mortality and health management.

The Mekong Delta in Vietnam is one of the most productive aquaculture regions in the world, in which the red tilapia (Oreochromis spp.) industry is a small-scale industry that mainly supplies local markets in the delta region. Little is known about the frequency of mortality events and health management in this sector. We describe red tilapia floating cage production systems in the Mekong Delta, Vietnam, for the purposes of quantifying mortality and associated production factors, and describing practices that may influence pathogen introduction and spread to and from farms. In July 2014, approximately 50 red tilapia farmers from 4 provinces (201 farmers in total) were randomly selected and interviewed. Median overall perceived mortality (PM) within a production cycle was 35%. Overall PM was found to be affected by province (p < 0.01), age of farmers (p = 0.01), anticipated main reason for PM in the first 2 wk (p = 0.03), most common market for the fish (p = 0.02), and whether farmers recorded stocking information (p = 0.01). Based on the interviews, we describe and discuss processes that potentially affect pathogen introduction and spread on these farms, such as movements of live and dead fish, distances between farms, mechanical transmission, and biosecurity practices such as treating fish before stocking, using disinfectants, and sharing equipment, and harvesters' movements. This study provides fundamental understanding of red tilapia aquaculture management in the Mekong Delta, and describes management factors that could become important in the event of disease outbreaks.

Survival of heterophyid metacercaria in common carp (Cyprinus carpio).

Heterophyidae are small intestinal trematodes that infect vertebrates worldwide. Common carp (Cyprinus carpio) is one of the most preferred freshwater fish species by consumers in Asia, the region where fish-borne trematodes like Heterophyidae are most prevalent. How long Heterophyidae survive in common carp is unknown. The objective of this study was to quantify survival of Heterophyidae in common carp after experimental exposure. Fish of 0.18 g were either used as controls or exposed to 250 heterophyid cercaria for 24 h. Control fish did not become infected. Percentage infection of exposed fish at 0-2 (n = 53), >2-10 (n = 15), >10-20 (n = 11), and >20-27 (n = 33) weeks post exposure was 98, 80, 100, and 100 % respectively. The number of metacercaria per fish did not significantly decrease (P = 0.19) during 27 weeks after exposure: exp [3.6200-0.0193 × weeks post exposure]. All developed metacercaria were identified as Haplorchis spp. It was concluded that Heterophyidae may persist in carp for a long time, implying that harvestable carp are a risk to human health.

Higher attack rate of fish-borne trematodes (Heterophyidae) in common carp fingerlings (Cyprinus carpio) at lower fish weight.

Fish-borne zoonotic trematodes (FZTs) can cause pathology in humans. Fish weight was reported as important risk factor for transmission from snail to fish. However, in fingerlings, the relation between fish weight and infection is unknown. Aim was quantifying the effect of fish weight on infection probability, attack rate, and metacercariae burden of FZTs in common carps (Cyprinus carpio) between 1 and 20 g. Fish were either used as controls (n = 66) or exposed to 250 parapleurolophocercous cercariae (n = 254). Fish weight was analysed as continuous explanatory variable or classified in four categories with average weights of 0.7 g (n = 116), 4.0 g (n = 58), 8.2 g (n = 57) and 14.2 g (n = 23). The inverse relation between percentage of fish with metacercariae and fish weight is reflected in lower percentages of infected fish at higher weights [%infected = 100/(1 + e([-2.02+0.15 fish weight (g)])); p < 0.01], i.e. 89 %, 85 %, 63 % and 61 %, respectively, in the four groups. Control fish did not get infected. Attack rates were 0.0087, 0.0073, 0.0040 and 0.0033 fish infected per cercariae, respectively; the first two attack rates being significantly higher than the latter two. Mean number of metacercariae per weight group was 5, 5, 2 and 1, respectively, (p < 0.05), with an inverse relation using weight as continuous explanatory variable [p < 0.01; number metacercariae = e(1.76-0.13fish weight(g))]. Concluding, an inverse relation exists between fish weight and probability of infection, attack rate and parasite burden in common carp fingerlings. Reducing transmission to fingerlings might be an effective intervention method to improve food safety, reduce the absolute amount of FZTs in the environment and eventually reduce incidence in humans.

Field evaluation of diagnostic sensitivity (DSe) and specificity (DSp) of common tests for amoebic gill disease (AGD) and complex gill disease (CGD) in cultured Atlantic salmon (Salmo salar) in Scotland using Bayesian latent class models.

Amoebic gill disease (AGD) and complex gill disease (CGD) are the most significant marine gill diseases in salmon aquaculture in Scotland. Little is published about diagnostic performance of tests to detect these diseases, making it difficult to interpret test results. We estimated diagnostic sensitivity (DSe) and specificity (DSp) of common tests for AGD (gross AGD score, qPCR for Neoparamoeba perurans, histopathology) and CGD (gross proliferative gill disease (PGD) score, gross total gill score, histopathology). Because specifications in our sampling protocol implemented to encourage consistency across the farms might affect diagnostic performance of histopathology (historically the reference standard for gill diseases), we used Bayesian latent class models without reference standard. Cases and non-cases were based on less, medium, and severe stringent case definitions, representing different cut-off levels for the different tests. Gross gill scores for both diseases were excellent in designating non-diseased fish, DSps were generally around 1. To detect CGD, DSe of gross total gill score and gross PGD score were between respectively 0.81 (0.73 - 0.91 lower to upper 95% credible interval) and 0.53 (0.46 - 0.64) for medium stringent case definitions, and to detect AGD the DSe for the gross AGD score was between 0.53 (0.48-0.57) and 0.14 (0.07 - 0.22) for respectively the less and severe stringent case definition. Thus, gross gill scores were medium to good in designating truly diseased fish, implying some false negatives are expected. For CGD the DSe for gross total gill scores were the highest, for AGD it was the qPCR test at a DSe of 0.92 (0.86 - 0.99). For both diseases, DSe was lowest for histopathology, e.g. 0.23 (0.16 - 0.30) for AGD and 0.1 (0.07 - 0.14) for CGD under medium stringent case definitions, perhaps due to collecting the second gill arch on the right rather than the worst affected arch, whilst PCR sampling and gross gill scoring included multiple (PCR) or all (gross scoring) gill arches. The diagnostic goals of these tests differ; gross gill scoring provides a low-cost presumptive diagnosis, PCR a non-lethal confirmation of the presence of a specific pathogen and histopathology provides information on the underlying aetiology of gill damage as well as the extent, severity, and chronology of gill disease. An effective gill health surveillance strategy is likely to incorporate multiple diagnostic tools used in a complementary manner.

Distribution of trematodes in snails in ponds at integrated small-scale aquaculture farms.

In integrated small-scale aquaculture farming, animal and human excreta maybe used as fish feed and pond fertilizer, thereby enhancing transmission of fish-borne zoonotic trematodes (FZTs) from final hosts, like humans, pigs and chickens, to snails. Areas within a pond could vary in trematode egg-load due to the immediate bordering land, and this might provide implications for control of these trematodes or sampling in field studies measuring FZT prevalence in snails. We therefore estimated the effect of bordering land use on prevalence and FZT burden in snails in different areas within small-scale aquaculture ponds. Nine sampling areas within a pond were assigned in six ponds. For each sampling area, about 120 Melanoides tuberculata snails were collected. Based on land use bordering a sampling area, these were categorized in 5 risk-categories: low-risk (road, rice planted in pond, agriculture, or middle of pond), human access point to pond, livestock sty (pigs or poultry), both human access point and livestock sty, and water connection to canal. In total, 5392 snails were collected. Percentages of snails with parapleurolophocercous cercariae varied between 6% in areas categorized as low-risk and areas with livestock sty only to 15% in areas with both human access point and livestock sty; only this 15% was significantly different from the prevalence in the low-risk category. Percentages of snails with xiphidio cercariae did not differ between risk-categories and varied between 5% and 10%. Mean snail size was 15.2mm, and was significantly associated with both the probability of infection as well as parasite burden. Very small differences in parasite burden were found at different land use areas; the maximum difference was about 11 cercariae. This study demonstrated only small differences between areas surrounding a pond on risk of snails to be infected with fish-borne trematodes within different pond areas. In field studies on FZTs in M. tuberculata snails in ponds, sampling from ponds can therefore be done without considering areas within ponds.

Development and validation of a two-step real-time RT-PCR for the detection of eel virus European X in European eel, Anguilla anguilla.

Eel virus European X (EVEX) is one of the most common pathogenic viruses in farmed and wild European eel (Anguilla anguilla) in the Netherlands. The virus causes a hemorrhagic disease resulting in increased mortality rates. Cell culture and antibody-based detection of EVEX are laborious and time consuming. Therefore, a two-step real-time reverse transcriptase (RT-)PCR assay was developed for rapid detection of EVEX. Primers and probe for the assay were designed based on a sequence of the RNA polymerase or L gene of EVEX. The real-time RT-PCR assay was validated both for use with SYBR Green chemistry and for use with a TaqMan probe. The assay is sensitive, specific, repeatable, efficient and has a high r²-value. The real-time RT-PCR assay was further evaluated by testing field samples of European eels from the Netherlands, which were positive or negative for EVEX by virus isolation followed by an indirect fluorescent antibody test. The real-time RT-PCR assay allows rapid, sensitive and specific laboratory detection of EVEX in RNA extracts from 10% eel organ suspensions and cell cultures with cytopathic effects, and is a valuable contribution to the diagnosis of viral diseases of eel.