Identification and validation of cold responsive microRNAs of <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> using high throughput sequencing

Cold stress is one of the major abiotic factors that influence the productivity and geographical distribution of many agriculturally important crops like Hevea brasiliensis. Cultivation of H. brasiliensis in India is being extended to northeastern regions, where low temperature during winter adversely affects its survival, growth, and productivity. Developing cold-tolerant genotypes is a primary requisite to maximize the productivity under such challenging environmental conditions. However, lack of methods for early evaluation of cold tolerance in the newly developed clones and the extensive time required for assessing their tolerance in the field are major constraints for clonal selection. The present study was initiated with an objective to identify and characterize cold stress responsive miRNAs from H. brasiliensis that show stronger association with cold tolerance. Next generation sequencing using Illumina HiSeq method revealed the expression of 21 and 29 conserved miRNA (from clone RRIM 600) families in cold-stressed and control samples, respectively. Forty-two novel miRNAs were identified from this study. Upon differential expression analysis, eight conserved miRNAs were found commonly expressed in both the samples. When expression analyses were performed subsequently with six selected miRNAs in two Hevea clones (viz. RRII 105 and RRIM 600), miR169 showed a strong association with cold tolerance. miRNAs such as miR482 and miR159 also exhibited association with cold tolerance. This study suggests the possibility of employing these miRNAs as markers for cold tolerance after validation in more number of genotypes with varying levels of cold tolerance.     

Seed development of Araucaria angustifolia: plant hormones and germinability in 2 years of seeds production

New Forests - During seed development, plant hormones are involved in processes such as the accumulation of reserves, cellular activity and physiological responses. The present study aimed to...     

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.     

Lippia alba (Verbenaceae) hydrolate as sedative of tambaqui (Colossoma macropomum) juveniles in simulated transport conditions

Lippia alba hydrolate (LAH) is produced as a secondary product in essential oil extraction. The objective of this study was to evaluate the use of LAH in a simulation of the transport of tambaqui (Colossoma macropomum) juveniles. A series of 17 hr simulations were performed using load densities of 30, 60 and 90 juveniles per L and four LAH concentrations of 0.4%, 1%, 2% and 5%, with a control of just water. Fish survival, temperature, dissolved oxygen, conductivity, pH, ammonia excretion, alkalinity water hardness and net Na⁺, K⁺, Cl^‐ fluxes, were all evaluated before and after the transport simulations. Mucous cell densities in the gills were also determined microscopically, post simulation. The results showed that addition of 5% LAH achieved better survival during the simulation than the water control. Dissolved oxygen was significantly lower with an increased load density. Also, conductivity, NH₄ excretion and net ionic fluxes (K⁺ and Cl^?) were lower with 5% LAH, although mainly where higher load densities were tested. Mucous cell density was lower in LAH concentrations of 2% and 5%, again with higher load densities. In view of these findings, it is suggested that 5% LAH is an effective sedative for use in the transport of tambaqui, mainly with high load densities. This work is the first study of LAH and highlights its potential applications in aquaculture management.     

A review of reported effects of pelagic longline fishing gear configurations on target,bycatch and vulnerable species

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Establishment of a proteomic reference map for the gastrocnemius muscle in the rabbit (Oryctolagus cuniculus)

In several laboratory and production species, the establishment of a proteome reference map of a specific tissue has been accomplished. The rabbit is widely used as both a production and experimental animal. A lot of physiology research involving the gastrocnemius muscle of rabbit is described, although no reference proteome map is available. In this work, the first reference map of the rabbit’s gastrocnemius muscle using 2D gel electrophoresis and the identification of proteins through peptide mass fingerprinting (PMF) was established. A total of 45 proteins were localized and identified with three major roles: cell structure and contractile apparatus; metabolic and cell defense proteins. A reference map of major proteins expressed is described enabling possible comparisons with other physiological studies.     

Methods for Cercospora coffeicola protoplast isolation and genetic transformation with the green fluorescent protein

Cercospora coffeicola is the causal agent of brown eye spot on coffee leaves. Although the disease has significant importance, few molecular studies have been done with C. coffeicola. Here we report a protocol for isolating protoplasts as well as development of a genetic transformation system using Green Fluorescent Protein. High yields of protoplasts (≈10⁸/ml) were obtained from mycelial cultures from five isolates of C. coffeicola. One isolate was transformed with a vector encoding hygromycin resistance and Green Fluorescent Protein. Out of 43 hygromycin-resistant transformants obtained, Green Fluorescent Protein was highly expressed in one (2.3 %).     

Integrated production of fish (pacu <Emphasis Type="Italic">Piaractus mesopotamicus</Emphasis> and red tilapia <Emphasis Type="Italic">Oreochromis</Emphasis> sp.) with two varieties of garnish (scallion and parsley) in aquaponics system

Aquaponics is emerging as an alternative for high-health food production. Being able to identify the technical viability of non-conventional plants and fish species would help to increase the interest and possibilities in aquaponic systems. The goal of the present study was to evaluate the aquaponics production of two garnish species: scallion (S) and parsley (P), using effluents of pacu and red tilapia culture. Two aquaponics devices were used, differing according to the fish species, generating two different effluents. Thus, for plant performance, four treatments were evaluated in a factorial design (plant species and fish effluent as main factors), as followed: Pacu-S, Tilapia-S, Pacu-P, and Tilapia-P, with three replicates each, for 35 days. Fish performance was evaluated using Student’s t test. Each experimental device included a fish tank, filters, and six experimental units for the plants (floating rafts). Results indicated that feed conversion ratio (FCR) was higher in tilapia as compared to pacu (p < 0.05); however, fish productivity and survival were similar between species. Plant performance parameters were similar with no significant differences regardless of the fish effluent (p > 0.05), except for higher number of leaves per plant in scallion cultured using pacu effluent. Plant performance comparing both plant species indicated that scallion performed better as compared to parsley in all parameters. In addition, scallion also performed better related to the plant quality index. The results indicate that pacu presented a viable alternative for the aquaponics production, and regarding to the garnish, scallion performed better results as compared to parsley.     

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.