Assessment of primers designed for the subspecies-specific discrimination among Babesia canis canis, Babesia canis vogeli and Babesia canis rossi by PCR assay

Canine babesiosis is an infectious disease caused by either Babesia gibsoni or Babesia canis protozoans. The latter is also classified under three different phylogenetic groups, referred to as subspecies B. canis canis, B. canis vogeli and B. canis rossi. The objective of the present study was to validate and standardize a PCR assay to discriminate the organisms at the subspecies level. First, the reference sequences of the 18S rRNA, 5.8S rRNA and 28S rRNA genes, including the internal transcribed spacer 1 (ITS1) and 2 (ITS2) of the most common species and subspecies of the genus Babesia were retrieved from the GenBank database. Subspecies-specific primers (BAB3, BAB4 and BAB5) and one genus-specific primer were designed from the alignment of the sequences. The PCR assays were evaluated in three different combinations of primer pairs in order to assure complete specificity for each reaction. The results of the tests had demonstrated effectiveness of the novel primer pairs BAB1/BAB3, BAB1/BAB4 and BAB1/BAB5 for the amplification of the subspecies-specific target fragments of 746 bp (B. c. canis), 546 bp (B. c. vogeli) and 342 bp (B. c. rossi) by PCR. The original enzymatic amplification assays with novel primers reported in this paper were confirmed to be a reliable tool for the specific discrimination among B. canis subspecies by single-step PCR assays.     

Digestible protein requirements and muscle growth in juvenile tambaqui (Colossoma macropomum)

Several investigations have been carried out to improve the productivity of tambaqui, an economically important fish species in Brazil and other Latin American countries. This study determined the digestible protein (DP) requirements in juvenile tambaqui by assessing their productive performance and nutritional efficiency. It also evaluated the effects of different dietary DP levels on the morphology and cellularity of skeletal fast muscle fibres. The 1750 tambaqui tested (6.53 ± 0.43 g body weight, 7.58 ± 0.18 cm length) were randomly distributed into 35 tanks. Fish were fed one of the seven isocaloric diets, which contained 140, 170, 200, 230, 260, 290 or 320 g/kg DP. The DP requirement, calculated by segmented (broken line) regression of weight gain data, was 290 g/kg. An increase in diet DP to 290 g/kg significantly improved final weight, weight gain, feed intake, specific growth rate and crude protein gain, and changed fibre diameter in deep muscle. Muscle fibres were randomly distributed into a mosaic pattern, characterized by fibres with different diameters. Treatments with 290 and 320 g/kg DP increased the frequency of large‐diameter fibres (>50 µm), indicating hypertrophic growth of skeletal muscle during the juvenile phase, which occurred in conjunction with hyperplasia.     

Gene pool and connectivity patterns of Pinna nobilis in the Balearic Islands (Spain,Western Mediterranean Sea): Implications for its conservation through restocking

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Use of Hydrodynamic Cavitation for Algae Removal: Effect on the Inactivation of Microalgae Belonging to Genus <Emphasis Type="Italic">Scenedesmus</Emphasis>

Excessive algae growth has generated conflicts on the use of water supplies; therefore, the focus on new technologies to remove algae from water bodies is demanding. The aim of the present study was to assess the effect of hydrodynamic cavitation on the inactivation of microalgae belonging to genus Scenedesmus. A laboratory-scale experimental apparatus was built in order to accomplish this goal; it consisted of a Venturi device designed to generate the cavitation phenomenon. Suspended microalgae samples were treated for 60 minutes under different cavitation intensities (cavitation number—Cv—ranging from 0.17 to 0.27). Results evidenced that microalgae decay over time can be modeled through first-order kinetics. The maximum removal efficiency (85%) was recorded at the highest cavitation intensity (Cv?=?0.17). The removal efficiency decreased as the cavitation number increased. Hydrodynamic cavitation was effective in inactivating Scenedesmus; it produced irreversible damages to cell morphology such as flotation spines removal, cell wall lesions, cytoplasm extravasation, and cavity formation. Assumingly, hydrodynamic cavitation has great potential to treat eutrophic water bodies. Furthermore, it represents a sustainable removal technique, since it does not produce secondary pollution.     

Finishing plant diet supplemented with microalgae meal increases the docosahexaenoic acid content in Colossoma macropomum flesh

The need for a sustainable aquaculture is increasing the use of plant ingredients in replacement to fishmeal and fish oil in diets for tambaqui Colossoma macropomum, which is leading to not detectable levels of docosahexaenoic acid (DHA) in its flesh. We evaluated the effect of a finishing plant diet supplemented with 5% of microalgae meal from Schizochytrium sp. (MD) on tambaqui growth, on proximal composition and fatty acid content of its flesh, comparing it to a non‐supplemented diet. One hundred and sixty‐two fish (489.67 g) were distributed into six tanks (2,000 L) and fed the experimental diets for a 90‐day period. Three fish per tank were euthanized for analyses every 15 days. The MD did not affect the growth and proximal composition of fish flesh. The MD increased the DHA content (from 14.81 to 38.60 mg/g of lipids) and the n‐3:n‐6 ratio (from 0.16 to 0.51) in the flesh of fish, beginning on the 15th day and reaching the highest DHA content on the 71st day (39.81 mg/g of lipids). We recommend C. macropomum to be fed with a finishing diet supplemented with microalgae meal for 71 days before slaughter to improve the DHA content and n‐3:n‐6 ratio in the flesh.     

Ion,Protein, Phospholipid and Energy Substrate Content of Oviduct Fluid During the Oestrous Cycle of Buffalo (Bubalus bubalis)

The aim of this research was to analyse the composition of oviduct fluid (ODF) in buffalo cows at different oestrous cycle phases to fulfil the requirements of buffalo embryos in vitro. ODF was collected by chronic cannulation from three cows that were synchronized by administering a synthetic prostaglandin. Based on hormonal profiles, the pre‐ovulatory, ovulatory, post‐ovulatory and luteal phases of the oestrous cycle were defined. The volume of ODF produced (ml/24 h) was influenced by the oestrous cycle, with values (mean ± SE) around ovulation (1.0 ± 0.2) greater (p < 0.05) than in both the luteal (0.4 ± 0.1) and the post‐ovulatory phases (0.5 ± 0.1), but not different from the intermediate values in the pre‐ovulatory phase (0.8 ± 0.2). Among cycle phases, no differences were found in sodium, potassium, calcium and magnesium concentrations (130.0 ± 1.1, 5.1 ± 0.3, 2.8 ± 0.1 and 0.59 ± 0.04 mmol/l respectively). Interestingly, the chloride secretion (μm /24 h) was higher (p < 0.05) at ovulation (150.2 ± 16.5) than during both the luteal (73.7 ± 22.0) and the post‐ovulatory phases (63.7 ± 11.2), with intermediate values in the pre‐ovulatory phase (113.4 ± 23.5). Glucose concentration (mmol/l) was higher (p = 0.056) in the pre‐ovulatory phase (0.06 ± 0.02) than in the luteal (0.02 ± 0.01) and post‐ovulatory (0.02 ± 0.01) phases but not different from values in the ovulatory phase (0.04 ± 0.02). Concentrations of pyruvate and lactate among oestrous cycle phases were similar (0.08 ± 0.01 and 1.0 ± 0.1 mmol/l respectively). The total quantity of phospholipids (μmol/24 h) was greater (p < 0.05) at ovulation (0.21 ± 0.02) compared with the luteal, pre‐ovulatory and post‐ovulatory phases of the cycle (0.09 ± 0.02, 0.13 ± 0.02 and 0.09 ± 0.01 respectively). No differences were found in either the protein concentration (1.8 ± 0.3 mg/ml) or the quantity of proteins secreted in 24 h (1.8 ± 0.4 mg) among oestrous cycle phases. In conclusion, this study provides the first characterization of buffalo ODF during the oestrous cycle, showing species‐specific differences that may be useful for developing suitable media for buffalo in vitro embryo production.     

On the Quantum Nature of the Shared Proton in Hydrogen Bonds

The relative influence of thermal and quantum fluctuations on the proton transfer properties of the charged water complexes H5O2+ and H3O2- was investigated with the use of ab initio techniques. These small systems can be considered as prototypical representatives of strong and intermediate-strength hydrogen bonds. The shared proton in the strongly hydrogen bonded H5O2+ behaved in an essentially classical manner, whereas in the H3O2- low-barrier hydrogen bond, quantum zero-point motion played a crucial role even at room temperature. This behavior can be traced back to a small difference in the oxygen-oxygen separation and hence to the strength of the hydrogen bond.