Broodstock conditioning of the Portuguese oyster (Crassostrea angulata,Lamarck, 1819): influence of different diets

The Portuguese oyster Crassostrea angulata shows great potential in oyster farming. The conservation of pure populations of this species is important for production diversification and biodiversity preservation. In this way, the zootechnological development for seed hatchery production is extremely important. Broodstock conditioning is a key step in the process of rearing bivalves in a hatchery. Many factors regulate the reproductive cycle, being food one of the most important ones. To evaluate the effect of different diets on C. angulata reproductive performance, broodstock were conditioned with different food regimes formulated fundamentally by flagellates (Diet 1 – Pavlova lutheri and Isochrysis galbana clone T‐ISO; Diet 2 – P. lutheri, T‐ISO and Skeletonema costatum) and constituted fundamentally by diatoms (Diet 3 – S. costatum and Chaetoceros calcitrans; Diet 4 – P. lutheri, S. costatum and C. calcitrans). During conditioning, samples of oysters were collected to evaluate condition index, gonadal development and biochemical composition. At the end of the conditioning period, oysters were induced to spawn to evaluate reproductive output (fecundity, fertilization rate and D‐larvae development). The diets had an impact on the gametogenesis process, energy storage and reproductive output performance, being the best results those obtained in broodstock fed with the diatoms‐predominant diets. However, those fed with diets majority flagellates had an unsuccessful performance. Holistic approaches incorporating all results in this study reveal and reinforce the idea that the diatom species used presented the nutritional requirements to C. angulata broodstock, being essential in the conditioning phase.     

Morphological and physiological studies on densely branched lateral roots triggered by localized phosphate in Sesbania cannabina

Densely branched lateral roots (DBLRs) in Sesbania cannabina are formed in response to patchily distributed phosphorus (P) in volcanic soils. Little attention has been paid to morphological and physiological responses of DBLRs. Here, we investigated the relation between plant growth and DBLR development, enzymatic activities involved in P acquisition, and the influence of arbuscular mycorrhizal fungi (AMF), which contribute to P uptake, to clarify the function of DBLRs. We investigated DBLR development induced by localized application of P fertilizer and we compared the activities of phosphoenolpyruvate carboxylase (PEPCase) and acid phosphatase (APase) between DBLRs and non‐DBLRs. Additionally, plants were grown with or without AMF to investigate the effect of AMF colonization on the numbers of DBLRs and plant P uptake, and we compared AMF colonization between DBLRs and non‐DBLR roots. Secondary to quaternary lateral DBLRs were produced after the primary lateral roots passed near P fertilizer. P_i content per DBLR increased as DBLRs developed, promoting higher shoot growth. Under P deficiency, PEPCase and APase activities increased in non‐DBLR, but were significantly lower in DBLRs in the same plants. AMF inoculation changed the root system architecture by significantly decreasing the number of DBLRs, and AMF colonization was lower in DBLRs than in non‐DBLRs. Our results indicate that DBLR formation is a P‐coacquisition strategy of S. cannabina grown in P‐deficient andosolic soil. Roots that form DBLR are clearly different from non‐DBLR roots in morphological and biochemical response and AMF symbiosis.     

Metagenomic analysis using 16S ribosomal RNA genes of a bacterial community in an urban stream,the Tama River,Tokyo

In an effort to determine genus- or species-level taxonomic profiles and diversity of bacterial consortia in the Tama River around urban Tokyo, next-generation sequencing technology targeting a 16S ribosomal RNA (rRNA) gene amplicon was employed. Metagenomic analysis performed by an Ion Personal Genome Machine after sequentially filtering samples through 5-, 0.8- and 0.2-μm filters yielded 1.48 Gb of 16S sequences (average 2.38 M reads/sample). The results indicated that half of the bacterial sequences belonged to Proteobacteria, followed by Bacteroidetes, Actinobacteria and Cyanobacteria. Flavobacterium (Bacteroidetes), possibly including a potential fish pathogen, was the most numerous genera in the Tama River metagenome, and accounted for?~?16% of assigned 16S reads, followed by Mycobacterium. Other dominant bacterial genera including Zoogloea, Sediminibacterium, Hyphomicrobium, Sphingopyxis, Thiothrix and Lysobacter, were thought to be associated with waste water and sludge. MiSeq metagenomic analysis revealed that environmental factors, particularly water temperature, influenced the bacterial composition throughout the year, with a strong negative correlation observed for Proteobacteria and a positive correlation for Bacteroidetes. In terms of bacterial genera, Flavobacterium was positively correlated with temperature, while Polaromonas, Pseudomonas and Bradyrhizobium were negatively correlated with this, suggesting dynamic change in the free-living bacterial population throughout the year and versatile adaptation strategies in relation to environmental factors.     

The decline of fisheries on the Madeira River,Brazil: The high cost of the hydroelectric dams in the Amazon Basin

Despite being considered beneficial by providing a clean and renewable source of energy, the construction of hydroelectric dams has extremely negative implications for Amazonian fisheries. This study investigated the impacts of the Santo Antônio and Jirau hydroelectric dams on the fishery stocks of the Madeira River. This investigation was based on fish catch data from the Z‐31 fishing colony, located in the municipality of Humaitá, in Amazonas State, Northern Brazil. Data were collected daily and provided information on the date of return from each trip, the fish species targeted, and the total catch (kg) between January 2002 and September 2017. The results indicated reductions of 39% in the mean annual catch and 34% in the mean monthly catches. These results highlight the high price paid by local fish communities for the development of hydroelectric power in the Amazon basin.     

Dietary Protein Hydrolysates from Animal By-Products: Digestibility and Enzymatic Activity for Dourado Salminus brasiliensis

Fishmeal (FM) is a high-cost and scarce feedstuff. Notwithstanding, the formulation and processing of diets for carnivorous fish strongly rely on FM as a protein source, given its palatability and biological value. Animal by-product hydrolysates are high-quality feedstuff and can substitute FM in aquafeeds. This study evaluated the digestibility of hydrolysates from tilapia residue (TR), tuna head (TH), swine liver (SL), and poultry liver (PL), and the profile of digestive enzymes in juvenile dourado, Salminus brasiliensis, a carnivorous Characin, fed diets containing graded levels of the hydrolysates. Inclusion of hydrolysates in diet formulations lowered the pH of feed but did not alter feed intake by fish. Higher apparent digestibility coefficients (ADCs) of diets were recorded for fish fed diets containing TR and SL, and lower ADCs were recorded for fish fed diets containing TH. Protease and lipase activities in the animal’s stomach were higher, especially for those fed with diets containing SL. Amylase activity was higher in pyloric caeca, whereas in the intestines, the higher activity was seen in fish fed control and TR diets. Animal by-product hydrolysates were highly digestible for dourado, and the enzymatic profile of fish was influenced by the nutrient contents of diets.     

Depletion study and estimation of withdrawal periods for florfenicol and florfenicol amine in pacu (Piaractus mesopotamicus)

The intensive production of farmed fish is at a global all‐time high, and the control of bacteria proliferation in fish farms requires the frequent use of antimicrobials. This practice raises important environmental concerns related to the emergence of antimicrobial‐resistant bacteria strains. Only a few antimicrobial drugs have been approved for use in aquaculture, one of which is florfenicol. This work studies the depletion and withdrawal period of florfenicol and its main metabolite, florfenicol amine, in pacu (Piaractus mesopotamicus), a neotropical characin widely farmed in the southern hemisphere. Juvenile pacu (average weight of 724 g) were stocked in a closed‐loop laboratory system with controlled water temperature (25.8°C), and were fed for 10 consecutive days with a diet containing an intended dose of 10 mg/florfenicol per kg bw. Muscle and skin tissues were collected at 1, 3, 6, 8, 10, 12 and 16 days post‐treatment, and florfenicol and florfenicol amine were quantified using a validated ultra‐high performance liquid chromatography‐tandem mass spectrometry (UPLC‐MS/MS) method. The limits of quantitation for florfenicol and florfenicol amine were 10 ng/g in muscle and 50 ng/g in skin. Considering a maximum residue limit of 1000 ng/g for the sum of florfenicol and florfenicol amine in muscle with skin in natural proportions a withdrawal period of 5 days (water temperature 25.8°C) or 129 degree days was calculated on the basis of the upper limit of the one‐sided 95% confidence interval for the 99^th percentile derived from the residue depletion study.     

Leucocyte profile and growth rates as indicators of crowding stress in pejerrey fingerlings (Odontesthes bonariensis)

Crowding is one of the most common stressors found in intensive aquaculture, compromising growth rates and immune function. Plasmatic cortisol is a classic stress biomarker for fish, but its quantification is expensive and demands blood volumes that small individuals do not provide, constraining the usage of this technique to assess stress in fingerlings. The leucocyte profile is an alternative methodology to quantify stress with reduced costs and volumes of blood. Stress conditions promote neutrophilia and lymphocytopenia as response to elevated glucocorticoids levels. Considering the difficulties to assess stress imposed by intensive fish farming using measurement of glucocorticoid hormones, this study aimed to evaluate the stress‐induced changes in leucocyte profiles and growth rates imposed by crowding in fingerlings of Odontesthes bonariensis, a promising South‐American candidate for freshwater aquaculture. To meet these objectives, fingerlings (initial weight 0.05 ± 0.06 g and length 1.68 ± 0.13 cm) were reared for 45 days under three rearing densities (1, 5 and 10 fingerlings L^?1). At the end of this period, fish were anaesthetized and euthanized to obtain the leucocyte profile, neutrophil:lymphocyte ratio (N:L) and growth rate. Increasing density promoted: significant reduction in growth (final length, weight and specific growth rate); neutrophilia and lymphocytopenia; and increased N:L ratio. Concluding, the tested rearing densities imposed distinct levels of stress characterized by different N:L ratio, demonstrating that the leucocyte profile is a reliable alternative to measure stress levels in O. bonariensis fingerlings and probably in other fish species.     

Hematoporphyrin-based photodynamic therapy for cutaneous squamous cell carcinoma in cats

Photodynamic therapy (PDT) using a haematoporphyrin derivative (Photogem®, General Physics Institute and clustes Ltda) as photosensitizer and light emitting diodes (LEDs) as the light source was evaluated in 12 cats with cutaneous squamous cell carcinoma. Lesions were illuminated with LEDs, (300 J/cm for 30 min) 24 h after the administration of the photosensitizer. Clinical responses were classified as complete disappearance of the tumour with total re-epithelialization; partial response (a reduction greater than 50%); and no response (less than 50% reduction). Tumours localized to the pinna treated with one ( n  = 3) or two ( n  = 4) applications of PDT yielded no response. Highly invasive tumours of the nose and nasal planum also showed no response, after two treatments ( n  = 2). A combination of PDT and surgery was performed in three cases. Two cats showed partial response and one complete response with one application of therapy 30 days after nasal surgery. Small and noninfiltrative lesions ( n  = 3) of the nasal planum showed a PR with one application ( n  = 2) and a CR with two applications ( n  = 1). This study shows that PDT using Photogem® and LEDs can provide local control of low-grade feline squamous cell carcinoma. The addition of PDT to surgery in more invasive cases may help prevent recurrence.