Single nucleotide polymorphisms in the growth hormone gene of Oreochromis niloticus and their association with growth performance

Polymorphisms in the growth hormone (GH) gene that is associated with the growth rate of farmed fish have been the target of many breeding programmes. The present study aimed to identify single nucleotide polymorphisms (SNPs) in GH gene regions to evaluate the association of SNP variations with the growth rate of two Nile tilapia: Oreochromis niloticus (Linnaeus, 1758) strains. The targeted regions were amplified, sequenced, aligned and screened for the presence of SNPs; thereafter, performance tests were used to check for the association between SNPs and weight. Allele and genotype frequencies were estimated for each SNP and genotype. Genotype blocks or sets of SNP genotypes and frequencies were also estimated. Association between SNPs and growth rate was statistically evaluated using a univariate linear mixed model that included both fixed and random effects. A total of 10 SNPs were identified, nine in the proximal promoter and one located in the 5′ UTR, forming 10 genotype blocks. In all weight recordings, five genotype blocks were significantly associated with the highest weights. Single nucleotide polymorphisms 6‐10 were also found to be significantly associated with growth (p‐value < .05). Genotypes with higher additive genetic values for weight were identified in the Chitralada strain, suggesting a possible impact of these additive effects of the GH SNP genotype on the growth rate of Nile tilapia. These findings may be used as part of marker‐assisted selection in tilapia breeding programmes.     

The effect of feeding commercial diets on the development of juvenile crucian carp (Carassius carassius,L.). Part 1: Skeletal deformations

The crucian carp (Carassius carassius, L.) is a cyprinid freshwater species, whose wild‐living populations have decreased in recent years due to an increasing competition of invasive species. Several initiatives were launched, attempting to reintroduce this fish back into its native open waters, which requires the use of crucian carp fry grown under controlled conditions. However, the use of popular commercial diets results in severe developmental abnormalities. The aim of this study was to analyse the impact of feeding natural feed (Chironomidae sp. larvae) or two popular commercial diets on the skeletal development of crucian carp juveniles. Whole‐mount staining and histological methods were used in the study. Skeletal tissues of fish fed both commercial diets were demineralized and displayed a variety of deformities, such as abnormal vertebral spines and vertebral fusion, as well as a new type of deformation, which involved the action of dorsal fin pterygiophores upon the spine. Finally, a novel analytical method was proposed, which utilizes LA‐ICP‐MS for the estimation of bone mineralization. The study proved that basic commercial diets are inadequate for the rearing of crucian carp juveniles.     

Uncertainty assessment of soya bean yield gaps using DSSAT‐CSM‐CROPGRO‐Soybean calibrated by cultivar maturity groups

Soya bean yield gap can be caused by different factors resulting in uncertainties when the objective is to use such information for farm decision‐making and reference yield determination. Thus, this study aimed to quantify the soya bean yield gap for four sites, located in Southern and Midwestern Brazil, as well as the uncertainties of that related to cultivars, sowing dates, soil types and reference yields. The crop simulation model DSSAT‐CSM‐CROPGRO‐Soybean was calibrated for cultivars with similar maturity groups, based on the data obtained from the best farmers at the county level. The yield gap by water deficit (YG_WD) was obtained through the difference between potential and attainable yields, and that one caused by sub‐optimum crop management (YG_CM) by subtracting actual yield of each county, obtained from official statistics between 1989/90 and 2014/15 growing seasons, from the estimated attainable yield. The yield was simulated using four sowing dates, three soil types and two soya bean maturity groups by county. The reference yield uncertainty was quantified using yield reference from crop model and regional winners of the soya bean yield context, conducted by CESB (Brazilian Soybean Strategic Committee), for the growing seasons from 2013/14 to 2015/16. The crop model showed a good agreement between measured and simulated crop development and growth using calibration by maturity group, with low root mean square error (347 kg/ha). Southern sites had a mean YG_WD of 1,047 kg/ha, while in the Midwest, it was lower than 100 kg/ha. The YG_CM was 1,067, 528, 984 and 848 kg/ha, respectively, for Castro, PR, Mamborê, PR, Montividiu, GO and Primavera do Leste, MT, representing the opportunity for yield gain when having the best farmers as reference. The maturity groups, sowing dates and soil types showed to be an important source of uncertainty for yield gap determination, being recommended to investigate the farms in detail for an appropriate quantification. The reference yield showed expressive uncertainties, with some farmers presenting conditions to increase their soya bean yields by more than 3,000 kg/ha, when considering as reference the yields obtained by the winners’ farmers. These results show that uncertainties must be reduced when assessing farm yield gaps, in order to ensure that expected rate of soya bean yield growth could be reached by adopting the same technologies from CESB winners and best farmers in the county as a reference.     

Limnology of Macrobrachium amazonicum grow‐out ponds subject to high inflow of nutrient‐rich water and different stocking and harvest management

We evaluated the water characteristics and particle sedimentation in Macrobrachium amazonicum (Heller 1862) grow‐out ponds supplied with a high inflow of nutrient‐rich water. Prawns were subject to different stocking and harvesting strategies: upper‐graded juveniles, lower‐graded juveniles, non‐graded juveniles+selective harvesting and traditional farming (non‐grading juveniles and total harvest only). Dissolved oxygen, afternoon N‐ammonia and N‐nitrate and soluble orthophosphate were lower in the ponds in comparison with inflow water through the rearing cycle. Ponds stocked with the upper population fraction of graded prawns showed higher turbidity, total suspended solids and total Kjeldahl nitrogen than the remaining treatments. An increase in the chemical oxygen demand:biochemical oxygen demand ratio from inlet (4.9) to pond (7.1–8.0) waters indicated a non‐readily biodegradable fraction enhancement in ponds. The sedimentation mean rate ranged from 0.08 to 0.16 mm day^?1 and sediment contained >80% of organic matter. The major factors affecting pond ecosystem dynamic were the organic load (due to primary production and feed addition) and bioturbation caused by stocking larger animals. Data suggest that M. amazonicum grow‐out in ponds subjected to a high inflow of nutrient‐rich water produce changes in the water properties, huge accumulation of organic sediment at the pond bottom and non‐readily biodegradable material in the water column. However, the water quality remains suitable for aquaculture purposes. Therefore, nutrient‐rich waters, when available, may represent a source of unpaid nutrients, which may be incorporated into economically valued biomass if managed properly.     

Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis

In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as follows: (1) sera from 34 animals with natural B. canis infection, confirmed by blood culture and PCR, as well as 51 sera samples from healthy dogs with negative results by the agar–gel immunodiffusion (AGID) test for canine brucellosis, were used as the control panel for B. canis infection; and (2) to scrutinize the possibility of cross reactions with other common dog infections in the same geographical area in Brazil, 60 sera samples from dogs harboring known infections by Leptospira sp., Ehrlichia canis, canine distemper virus (CDV), Neospora caninum, Babesia canis and Leishmania chagasi (10 in each group) were included in the study. The ELISA using heat soluble bacterial extract (HE-antigen) as antigen showed the best values of sensitivity (91.18%), specificity (100%) and accuracy (96.47%). In the WB analyses, the HE-antigen showed no cross-reactivity with sera from dogs with different infections, while the B. canis sonicate had various protein bands identified by those sera. The performance of the ELISA standardized with the heat soluble B. canis antigen indicates that this assay can be used as a reliable and practical method to confirm infection by this microorganism, as well as a tool for seroepidemiological studies.     

Intestinal antiinflammatory activity of a lyophilized infusion of Turnera ulmifolia in TNBS rat colitis

Turnera ulmifolia is a plant popularly known in Brazil and South America as chanana. Some species of Turnera are widely used in folk medicine for different types of inflammatory diseases. In this study, the preventive intestinal antiinflammatory activity of a lyophilized infusion obtained from the aerial parts of T. ulmifolia was tested in the trinitrobenzenesulphonic acid (TNBS) model of rat colitis. The results obtained revealed that pretreatment to colitic rats with the extract, at 250 and 500 mg/kg, significantly attenuated the colonic damage induced by TNBS. This beneficial effect was associated with an improvement in the colonic oxidative status, since the infusion prevented the glutathione depletion that occurred as a consequence of the colonic inflammation. On the other hand, this antioxidant activity was confirmed in in vitro studies. In conclusion, the preventive effect exerted by the lyophilized infusion of T. ulmifolia in the TNBS model of rat colitis is probably related to its antioxidant properties, due to its flavonoids content.     

Proteomic analysis of membrane preparations from developing Pinus radiata compression wood

For coniferous gymnosperms, few data exist as to the contribution of the membrane-associated proteome to cell wall and wood formation. In this study, we begin to address this knowledge deficiency by examining the proteomic profile of Golgi-enriched membrane preparations derived from developing Pinus radiata compression wood. These membrane preparations were generated by a combination of discontinuous sucrose gradient centrifugation and Triton X-114-based phase separation. Fractionation by phase separation removed contaminating proteins associated with the cytoskeleton and enabled the discrimination between soluble and membrane-bound/integral proteins. The proteomic analysis of the resulting aqueous and detergent phases using high-performance liquid chromatography-tandem mass spectrometry resulted in the identification of 175 proteins. The majority of the identified proteins were membrane bound/integral and originated from cellular components such as the nucleus, plastids, endoplasmic reticulum, plasma membrane and Golgi vesicles. On the basis of bioinformatic analysis, many of the identified proteins were predicted to be involved either in the regulation of wood formation or in cell wall biosynthesis, which indicated that the proteomic analysis of non-cytosolic proteins in developing xylem is a useful strategy to investigate the molecular aspects of wood formation in pine.     

Cryopreservation of yellow-poplar and sweetgum embryogenic cultures

Cryopreservation has become anessential tool for operational application offorest tree embryogenic cultures, due to thelong evaluation periods needed for treesregenerated from these cultures. Fiveyellow-poplar (Liriodendron tulipifera)and seven sweetgum (Liquidambar spp.)embryogenic culture lines werestored in liquid nitrogen for 48 hours, afterwhich they were thawed and tested for regrowthand ability to produce somatic seedlings.Combinations of two sorbitol pretreatments andthree dimethylsulfoxide (DMSO) cryoprotectantlevels were evaluated for their impact onrecovery following cryogenic storage. The bestresults were obtained with 0.4 M sorbitol and5% DMSO, which provided 100% recovery.Somatic seedlings were regenerated from allculture lines and treatments, except for atransgenic sweetgum line.     

Specific hybridization real‐time PCR probes for Phytophthora ramorum detection and diagnosis

Sudden Oak Death, caused by Phytophthora ramorum, poses a serious threat to native American oaks, and is also present in Europe where it has been isolated from numerous European ornamental plant nurseries. Its proven aggressiveness against plants in the Fagaceae and Ericaceae and the damage it has caused in North America have lead to it being assigned quarantine status. The timely and accurate detection of P. ramorum is a critical aid in the study of the epidemiology and biology of this pathogen. As a regulated organism, the availability of a sensitive and reliable assay is essential when attempting to achieve early detection of the pathogen. In this work, new specific hybridization probes for a real‐time PCR amplification method were found to be rapid, robust and labour‐saving, and proved suitable for routine use in a molecular diagnostic laboratory.