Short-term dietary supplementation with the microalga Parietochloris incisa enhances stress resistance in guppies Poecilia reticulata

Two trials were conducted to determine the effects of dietary enrichments with the microalga Parietochloris incisa , rich in arachidonic acid (ARA), on stress resistance in guppies Poecilia reticulata . The microalga was added to commercial diets as a neutral lipid (NL) extract and its fractions or as broken cells. Experimental diets were applied for a period of 14 days. In trial 1, commercial diets were supplemented with NL (containing 25 mg ARA and 0.11 mg β-carotene g⁻¹ feed), its triacylglycerol (TAG) fraction (containing 25 mg ARA g⁻¹ feed and no β-carotene) and the β-carotene fraction (containing 0.11 mg carotenoid g⁻¹ feed and minute amounts of ARA). Neutral lipid-fed fish demonstrated the highest resistance ( P <0.05) to osmotic stress (32-ppt NaCl), followed by fish fed with diets supplemented with TAG and β-carotene alone, which were more resistant than control ( P <0.05). In trial 2, fish fed diets supplemented with higher levels of broken alga (26.1 mg ARA g⁻¹ feed) were more resistant ( P <0.05) to stress as compared with fish fed lower ARA (16.3 mg g g⁻¹) or an unsupplemented control diet. We suggest a dietary supplementation with broken P. incisa cells to enhance stress resistance in guppies before a stressful event.     

Environmental and physiological conditions affecting Tetrahymena sp. infection in guppies, Poecilia reticulata Peters

Parasitic infections caused by Tetrahymena sp. constitute a serious problem in guppies, Poecilia reticulata. Tetrahymena was isolated from skin lesions of naturally infected guppies in a commercial aquaculture farm, cultured in vitro and used in subsequent experimental infections. In addition to guppies, angelfish, Pterophyllum scalare, platyfish, Xiphophorus maculates, and neontetra, Paracheirodon innesi, were susceptible, whereas tilapia (Oreochromis niloticus xO. aureus) was resistant. The ciliate had a high affinity for dead fish. Skin abrasion did not affect the infection, but fish with gas bubble disease exhibited a significantly higher infection than non-affected fish. Infection was significantly higher when fish were exposed to high levels of ammonia, high organic load and low water temperatures. Under shipment conditions, infection was significantly elevated. Full recovery was achieved at a low fish density. Results suggest that poor environmental and physiological conditions enhance infection with Tetrahymena sp.     

Simultaneous effect of sex and dose on efficacy of clove oil,tricaine methanesulfonate, 2‐phenoxyethanol and propofol as anaesthetics in guppies,Poecilia reticulata (Peters)

We studied the simultaneous effect of sex and dose on anaesthesia efficacy to estimate, if possible, the lowest effective dose (LED) for clove oil, tricaine methanesulphonate (MS‐222), 2‐phenoxyethanol (2‐PE) and propofol in mature guppies. LED is the lowest dose needed to reach A5 stage in a mean time of 3 min, with mean recovery (R5) time of 5 min. We used four doses/anaesthetic: 25, 50, 75 and 100 mg/L for clove oil; 120,140,160 and 180 mg/L for MS‐222; 800, 1,000, 1,200 and 1,400 mg/L for 2‐PE, and 7.50, 8.25, 10.00 and 11.25 mg/L for propofol. Each dose was tested on 10 females and 10 males. Morbidity, mortality and behavioural changes were checked on two control groups (10 males and 10 females/group). Sedation (A3), A5 and R5 times were recorded. Significant interactive effect dose*sex on A5 time was found for each anaesthetic agent (p_dose*sex < .05). Except for MS‐222 (p_dose*sex = .284), significant interactive effect dose * sex on R5 time was found (p_dose*sex < .05). A5 time in females tended to be greater than in males, but, in general, R5 times were longer in males. Body size differences between males and females could explain these differences in MS‐222 on A5 time and for clove oil, 2‐PE and propofol on R5 time. No dose simultaneous meet LED′s conditions relating to both A5 and R5 times; therefore the lowest doses inducing A5 in a mean time of 3 min could be a safe guideline for anaesthetic procedure in both male and females.     

First record of freshwater fish on the Cape Verdean archipelago

The Cape Verdean islands form a distinct aquatic freshwater ecoregion characterized mainly by temporal water bodies with an adapted invertebrate community. Freshwater fish were not previously recorded from the archipelago. During a non-exhaustive survey of freshwater bodies on five islands of the archipelago, the first presence of a freshwater fish was recorded. Using barcoding sequences, the species was identified as the guppy (Poecilia reticulata), a highly invasive species alien to the Cape Verdean Islands.     

Dietary supplementation with the microalgae Parietochloris incisa increases survival and stress resistance in guppy (Poecilia reticulata) fry

Dietary enrichments with the arachidonic acid (ARA)‐rich microalga, Parietochloris incisa, on the survival of guppy (Poecilia reticulata) fry were examined. Diets were applied via Artemia enrichment to fish from two commercial farms for 34 and 36 days of experimental period (trials 1 and 2, respectively). In trial 1, Artemia nauplii were enriched with dry biomass of whole algal cells at 0 (control), 0.1, 0.2 and 0.4 mg mL^?1. Fry fed with Artemia enriched with 0.4 mg mL^?1 demonstrated the lowest mortality rates (24% and 1% in farms 1 and 2, respectively) compared with controls (36% and 13% in farms 1 and 2, respectively). In trial 2, fry were fed with Artemia, enriched with whole algal cells (0.4 mg ml^?1), algal hexane extract (HE; containing primarily ARA‐rich triacylglycerols and β‐carotene; 0.19 mg ml^?1) or the extraction residue (0.28 mg ml^?1). Acute stress (5 min air exposure) was applied after 18 days. The lowest mortality was recorded in the whole alga‐fed group (av. 26% and 2.6% in farms 1 and 2, respectively), with a slightly, but not significantly higher mortality in the HE‐fed group (av. 29% and 6.2% in farms 1 and 2, respectively). Elevated lysozyme was associated with the reduced mortality. Overall, the use of P. incisa as a dietary supplement for guppy fry during their first month of life enhanced their survival and stress resistance.     

Size‐based sexual dimorphism of nitrogen excretion in livebearing fishes

Nitrogen (N) and phosphorus (P) are important limiting or co‐limiting nutrients in many aquatic ecosystems. Consumers such as fishes can significantly impact the balance and redistribution of these nutrients through consumer‐driven nutrient recycling. Intraspecific variation in nutrient excretion rates can therefore have significant ecosystem impacts. Among individuals of sexually dimorphic consumers, variation in population size structure and sex ratio could potentially have impacts of similar magnitude. We tested for the effects of body size and sex on consumer‐driven nutrient recycling by measuring N and P excretion rates from eight species of poeciliid fishes. We found a strong positive effect of size on N excretion rates, as has been previously described among species. However, we found no effect of size on P excretion rates, nor did we find any difference in N or P excretion rates between sexes. Our work provides a preliminary analysis of how sexual dimorphism can lead to disparate nutrient excretion rates within consumer populations. These results indicate that variation in population sex ratios of sexually dimorphic consumers could have impacts at the ecosystem scale.     

Use of decapsulated Artemia cysts in ornamental fish culture

Two series of feeding experiments were conducted to study the feasibility of using decapsulated Artemia cysts for direct feeding to ornamental fish. The first series evaluated the dietary values of the dried and the brine decapsulated Artemia cysts against two conventional live feeds, Artemia nauplii and Moina for adults and fry of the guppy, Poecilia reticulata Peters. In the second series, brine cysts were used for feeding to fry of four other important ornamental fish species, viz. platy, Xiphophorus maculatus (Günther), swordtail, X. helleri (Heckel), molly, P. sphenops Cuvier & Valenciennes and black neon tetra, Hyphessobrycon herbertaxelrodi Géry, and the results were compared with those fed Moina. Evaluation of the performance of the fish fed the various diets was based on stress resistance, growth and survival of the fish. Our findings indicated that decapsulated cysts could be used as a substitute for Artemia nauplii or Moina in freshwater ornamental fish culture. Apart from being a hygienic off‐the‐shelf feed, the direct use of the cysts also signifies a new area of application for low‐hatch cysts in the ornamental fish industry, with concomitant saving in feed costs.     

Treatment development for systemic Tetrahymena sp. infection in guppies,Poecilia reticulata Peters

Antibacterial and antiparasitic agents and a cysteine protease inhibitor (E‐64) were tested against Tetrahymena infection, a serious problem in guppy production worldwide. Chemicals were tested in vitro by a colorimetric assay for Tetrahymena survival. The most effective were niclosamide, albendazole and chloroquine, with 23%, 35% and 60% survival, respectively, following 2‐h exposure to 100 ppm. Longer incubation periods resulted in greater reductions in survival. Niclosamide was further studied in vivo at different dosages, administered orally to Tetrahymena‐infected guppies. Mortality rates were significantly lower in all treatment groups; in trial I, 30% and 33% mortality in 5 and 40 mg kg^?1 niclosamide‐fed fish vs. 59% mortality in controls; in trial II, 35%, 13% and 10% in 50, 100 and 200 mg kg^?1 niclosamide‐fed fish vs. 64% in controls. The effect of the cysteine protease inhibitor E64 was tested in tissue culture, by measuring histolytic activity of the parasite (Tet‐NI) on a guppy‐fin cell line, based on cell depletion. Tet‐NI feeding activity was significantly reduced following pretreatment with E‐64 relative to non‐treated Tet‐NI. E‐64‐pretreated Tet‐NI was injected i.p. into guppies: recorded mortality rates were significantly lower (35%) than that in non‐treated Tet‐NI (60%), suggesting inhibition of the parasite’s cysteine protease as a possible therapeutic approach.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

Effect of feeding guppy fish fry (Poecilia reticulata) diets in the form of powder versus flakes

The effects of feeding guppy (Poecilia reticulata) fry a diet offered as either powder or flakes on their growth and survival were tested over a period of 8 weeks. The results show that the growth of both male and female fish was considerably enhanced when the diet was presented in the form of a finely ground powder compared with a flake form. Final average weights of fish given a diet containing the exact same ingredients (44.9% protein and 6.1% fat), from the same batch of raw materials in the powdered form were 280.0±12.1 mg compared with 114.6±19.9 mg for the diet given in the form of flakes. In a diet that had a higher fat level (45.1% protein and 10.6% fat), the difference in final weight attained was even more dramatic: 303.9±16.7 mg for the powder‐fed fish compared with 92.6±12.5 mg for the flake‐fed fish. The coefficient of variance in the fish fed a flake diet was significantly higher than that exhibited in the treatments fed the powder feed. The ability of the fish to consume the food in a rapid manner, preventing leaching of vital nutrients from the feed before being engulfed by the fish probably led to the better growth results exhibited by the fish given the powdered food.