Growth and gut morphology of diploid and triploid juvenile Atlantic cod (Gadus morhua)

The objective of this paper was to compare the growth and gut morphology of juvenile diploid and triploid Atlantic cod (Gadus morhua) reared under similar conditions. Individually tagged 36‐week‐old diploid (mean weight 49.3 ± 13.8 g) and triploid (mean weight 43.6 ± 11.2) juvenile cod were measured at intervals during a 29‐week growth trial. Data for weight, length, condition factor (K), hepato‐somatic index (HSI), gonado‐somatic index (GSI), Relative Gut Length (RGL) and pyloric caeca number were collected and results were analysed in relation to ploidy status, gender and family contribution. At the end of the experiment, only one family (M2xF3) had many representatives with a relatively even distribution of sexes and ploidies. Diploid females were significantly heavier and had higher K than triploid females in the M2xF3 family (body weight 371.2 ± 120.2 vs. 298.4 ± 100.7 g; K 1.1 ± 0.1 vs. 0.93 ± 0.1), but no differences were found between diploid and triploid males. In the other families (pooled data), no differences in body weight were found between the ploidy groups. In general, triploids had a shorter intestine (RGL) and fewer pyloric caeca than their diploid siblings regardless of gender suggesting possible impairments in nutrient utilization and growth.     

Resistance gene analogue isolation and RGA-based marker development for identifying downy mildew resistance in radish (Raphanus sativus L.)

Resistance Gene Analogs (RGAs)-based molecular markers can significantly enhance the breeding efficiency for disease resistance in various crops. However, RGAs isolation and RGA-based marker development have not been conducted in radish. In this study, two prevalent approaches, PCR amplification with degenerate primers and data-mining of radish EST databases, were used to isolate radish resistance gene candidate sequences. A total of 26 RsRGAs with high homology to known R-genes or RGAs were obtained by PCR amplification. Meanwhile, 257 R-gene-like unigenes/ESTs were identified using data-mining method. In total, 115 out of 124 designed RGA-specific primer pairs can successfully amplified the target bands. To explore the RGA genetic variability, 32 radish accessions were genotyped and 269 RGA loci were successfully identified. It was found that 44 RGA primers only generated monomorphic band and the remaining 71 RGA primers generated 225 RGA polymorphic bands among the 32 radish accessions. Based on RGA marker analysis, 32 radish accessions were clustered into four main groups at the similarity index of 0.68, which was in a high accordance with disease index from a downy mildew evaluation. This study might be the first report on candidate R-genes identification and RGA-based markers development in radish. These findings will facilitate the disease resistance identification and speed up the genetic improvement of DM resistances in radish breeding programs.     

Cryptosporidium species detected in calves and cattle in Dagoretti,Nairobi, Kenya

A total of 1,734 cattle faecal samples from 296 dairy-keeping households were collected from urban settings in Nairobi, Kenya. Modified Ziehl–Neelsen staining method and an immunofluorescence assay were used to identify those samples with Cryptosporidium oocyst infection. Oocysts from positive faecal samples were isolated by Sheather's sucrose flotation method and picked from the concentrate using cover slips. Genomic DNA was extracted from 124 of the faecal samples that were positive for Cryptosporidium and was used as template for nested PCR of the 18S rRNA gene. Twenty-five samples (20 %) were PCR-positive for Cryptosporidium, and 24 of the PCR products were successfully cloned and sequenced. Sequence and phylogenetic analysis identified 17 samples (68 %) as Cryptosporidium parvum-like, four samples (16 %) as Cryptosporidium ryanae, three samples (12 %) as Cryptosporidium andersoni and one sample (4 %) as Cryptosporidium hominis. To the best of our knowledge, this is the first genotyping study to report C. parvum-like, C. andersoni and C. hominis in cattle from Kenya. The results of this study show Cryptosporidium infections in calves and cattle may be potential zoonotic reservoirs of the parasite that infects humans.     

Intra and inter-laboratory reproducibility of a monoclonal antibody cocktail based ELISA for detection of allergen specific IgE in dogs: Proficiency monitoring of macELISA in six laboratories

The purpose of this study was to evaluate the reproducibility of results yielded using a monoclonal antibody based ELISA for detection of allergen specific IgE when run in six separate affiliated laboratories. On two separate occasions, duplicate samples of 15 different sera pools were independently evaluated by each laboratory in a single blinded fashion. The average intra-assay variance among reactive assay calibrators in all laboratories was 6.2% (range 2.6-18.2%), while the average intra-laboratory inter-assay variance was 12.1% (range 8.0-17.1%). The overall inter-assay inter-laboratory variance was consistent among laboratories and averaged 15.6% (range 15.1-16.6%). All laboratories yielded similar profiles and magnitudes of responses for replicate unknown samples; dose-response profiles observed in each of the laboratories were indistinguishable. Considering positive/negative results, inter-assay inter-laboratory concordance of results exceeded 95%. Correlation of OD values between and among all laboratories was strong (r>0.9, p<0.001). Correlation of OD values between the two separate evaluations was also high for all allergens except olive, which was attributed to lot-to-lot differences of allergen coated wells. Collectively, the results demonstrated that the monoclonal antibody based ELISA for measuring allergen specific canine IgE is reproducible, and documents that consistency of results can be achieved not only in an individual laboratory, but between laboratories using the same monoclonal-based ELISA.     

Flow cytometry: a quick method to determine ploidy levels in honeybush (<Emphasis Type="Italic">Cyclopia</Emphasis> spp.)

Cyclopia (honeybush) species are widely-used in the production of a South African herbal tea and are endemic to the fynbos region of the Western and Eastern Cape Provinces. Honeybush is still one of the orphan agriculture crops and recent breeding efforts by researchers are hampered by the lack of basic genetic information e.g. basic chromosome numbers and ploidy levels. This study determined nuclear DNA content and ploidy level of various genotypes of three Cyclopia species using flow cytometry and cytological counting of chromosomes. Nuclei analysis of young leaves of C. genistoides, C. longifolia and C. subternata were done using a flow cytometer, while root tip squashes were carried out in order to correlate flow cytometry results. Flow cytometry analysis indicated differences in the nuclear DNA content among and within species whilst the DNA ploidy level only differed among species. Cyclopia genistoides had a higher DNA ploidy level (≥?10C) and DNA content (10.63 pg) than C. longifolia (6.09 pg) and C. subternata (5.99 pg), with no differences observed between the ploidy level of the latter two species (6C). The inferred ploidy level from nuclear DNA content by flow cytometry was consistent in all 30 genotypes of C. longifolia, and 24 of the 25 C. subternata and in only four of the 15 C. genistoides studied genotypes. These findings are important in breeding new cultivars with desired horticultural traits, thus improving the commercial characteristics for the sustainable production of honeybush.     

Development of wheat kernels with contrasting endosperm texture characteristics as determined by magnetic resonance imaging and time domain-nuclear magnetic resonance

Spikes and seeds from diploid ‘einkorn’ wheat Triticum monococcum and two near-isogenic hard and soft common wheat (Triticum aestivum) lines were harvested at regular intervals from 7 days post-anthesis (dpa) and analysed by non-destructive magnetic resonance imaging (MRI) and time domain-nuclear magnetic resonance (TD-NMR). A large amount of free water occurred in rachises, glumes and awns of spikes collected at 7 dpa, and accumulated in the physiologically active cells of the endosperm at 21 dpa. In the final stages of kernel development, awns and seed embryos exhibited a high MR signal due to the presence of free water likely associated with biological activities. TD-NMR relaxation time distributions obtained by discrete exponential fitting, distributed exponential fitting and SLICING multivariate analysis offered detailed information on mobility behaviour of water molecules in developing seeds and were able to differentiate two soft and hard isolines from common wheat cv. Enesco at early stages of seed development.     

Why Certify? Motivations,Outcomes and the Importance of Facilitating Organizations in Certification of Community-Based Forestry Initiatives

Despite documented challenges, many community-based forestry (CBF) initiatives pursue forest certification. This study asked community-based forestry practitioners in Vermont what influenced their decisions to seek or not seek certification and what outcomes were realized from certification. Relationships, public image, value alignment and feedback on management practices were most commonly cited as both motivations for and results of certification. Expectations for economic benefits were low and price premiums for products were only occasionally realized. Informants complained of the increasing cost, complexity and time commitment required of certification. Overall, however, certified CBF informants felt certification was worth the expense. Group certificates and external funding significantly reduced certification costs to grassroots CBF initiatives. This study highlights the importance of facilitating organizations that can provide outreach, secure funding, understand the rules, handle documentation and develop markets for certified products.     

A wide diversity of zoonotic intestinal parasites infects urban and rural dogs in Neuquén,Patagonia, Argentina

The presence of parasites was investigated by the examination of 1944 dog faecal samples collected from urban (n = 646) and rural (n = 1298) areas of the province of Neuquén, Patagonia, Argentina. Parasitic agents (PA) were found in 37.86% of samples. A total of 15 different PA were detected, including Toxocara canis (16.35%), Taenia spp./Echinococcus spp. (12.65%), Trichurisvulpis (6.06%), Giardia spp. (1.29%), Toxascaris leonina (0.56%), Ancylostomacaninum (0.41%), Dipylidium caninum (0.31%), Diphyllobothrium spp. (0.10%), among others. Several of these PA are recognized as zoonotic agents. Therefore, the results of this investigation revealed that local population is exposed to a broad spectrum of zoonotic parasites by means of environmental contamination with dog faeces. Prevalence of PA was slightly higher in rural (40.06%) than in urban (33.44%) locations. Distribution of groups of PA (cestodes, nematodes, and protozoa) showed statistical differences between both habitats. Prevalence of cestodes (18.18%) and protozoa (11.86%) was significantly higher in the rural environment than in urban areas and nematodes (29.10%) were more frequent in urban locations. Infection of dogs with Linguatula serrata and Cryptosporidium sp. was demonstrated for the first time in Neuquén. Rural dogs of the study area are under hydatic disease control program, which includes treatment with praziquantel every 6 weeks; thus, the finding of high level of cestode infection in these areas is of great relevance. The epidemiology of zoonotic parasitic infections in urban and rural dogs showed different patterns and, in consequence, different control measurements should be applied in each location.     

Structural and biochemical characterization and evolutionary relationships of the fatty acid-binding protein 10 (Fabp10) of hake (<Emphasis Type="Italic">Merluccius hubbsi</Emphasis>)

A fatty acid-binding protein (FABP) from the liver of Argentine hake (Merluccius hubbsi) was isolated and characterized and its expression analyzed. The determination of its partial primary structures (72 %) showed that it presents highest identity with Fabp10, commonly termed liver basic-type FABP. The evolutionary tree showed greater relationship between the Fabp10 of hake (Me Fabp10) and the Fabp10 and the Fabp10a of teleost fish. Me Fabp10 had low affinity for palmitic, oleic and palmitoleic acid and high affinity for bilirubin, lysophosphatidylcholine and lysophosphatidylethanolamine, all of them important in the metabolic functions of the liver. Me Fabp10 was able to bind only one cis-parinaric acid molecule and was found to be expressed only in the liver.