Hydrology influences breeding time in the white-throated dipper

While nitrogen and carbon stable isotope values can reflect ecological segregation, prey choice and spatial distribution in seabirds, the interpretation of bulk stable isotope values is frequently hampered by a lack of isotopic baseline data. In this study, we used compound-specific isotope analyses of amino acids (CSIA-AA) to overcome this constraint and to study interspecific differences, seasonal and historical changes in trophic positions of five seabird species, three penguins and two petrels, from a sub-Antarctic seabird community. CSIA-AA allowed comparing trophic positions of seabirds with temperate and polar distributions. Gentoo and Magellanic penguins had the highest trophic positions during the breeding season (3.7 and 3.9), but decreased these (2.9 and 3.3) during the feed-up for moult. Intra-specific differences were also detected in Thin-billed prions, where carbon isotope values clearly separated individuals with polar and temperate distributions, both in the breeding and interbreeding periods. Thin-billed prions that foraged in polar waters had lower trophic positions (3.2) than conspecifics foraging in temperate waters (3.8). We further investigated historical changes by comparing museum samples with samples collected recently. Our pilot study suggests that Rockhopper penguins, Magellanic penguins and Thin-billed prions with temperate non-breeding distributions had retained their trophic levels over a 90–100 year period, while Gentoo penguins and Thin-billed prions with polar non-breeding distributions had decreased trophic levels compared to historical samples. In contrast, Wilson’s storm-petrels had slightly increased trophic levels compared to samples taken in 1924–1930. We applied compound-specific stable isotope analyses across a range of contexts, from intra-specific comparisons between stages of the breeding cycle to inter-specific seabird community analysis that would not have been possible using bulk stable isotope analyses alone due to differences in isotopic baselines.     

Development and Characterization of Novel Polymorphic Microsatellite Markers for Tapinoma indicum (Hymenoptera: Formicidae)

Tapinoma indicum (Forel) (Hymenoptera: Formicidae) is a nuisance pest in Asia countries. However, studies on T. indicum are limited, especially in the field of molecular biology, to investigate the species characteristic at the molecular level. This paper aims to provide valuable genetic markers as tools with which to study the T. indicum population. In this study, a total of 143,998 microsatellite markers were developed based on the 2.61 × 10⁶ microsatellites isolated from T. indicum genomic DNA sequences. Fifty selected microsatellite markers were amplified with varying numbers of alleles ranging from 0 to 19. Seven out of fifty microsatellite markers were characterized for polymorphism with the Hardy–Weinberg equilibrium (HWE) and linkage disequilibrium (LD) analysis. All seven microsatellite markers demonstrated a high polymorphic information content (PIC) value ranging from 0.87 to 0.93, with a mean value of 0.90. There is no evidence of scoring errors caused by stutter peaks, no large allele dropout, and no linkage disequilibrium among the seven loci; although loci Ti-Tr04, Ti-Tr09, Ti-Te04, Ti-Te13, and Ti-Pe5 showed signs of null alleles and deviation from the HWE due to excessive homozygosity. In conclusion, a significant amount of microsatellite markers was developed from the data set of next-generation sequencing, and seven of microsatellite markers were validated as informative genetic markers that can be utilized to study the T. indicum population.     

Production of an Antibody Fragment (scFv) Targeting PcrV Protein of Pseudomonas aeruginosa in Fed-Batch Cultivation Mode

Background: Pseudomonas aeruginosa is one of the opportunistic pathogens causing frequent hospital-acquired life-threatening infections in mechanically ventilated patients. The most significant virulence factor of P. aeruginosa is T3SS. PcrV is an important structural protein of the T3SS. Methods:In the current investigation, a recombinant scFv mAb against the PcrV protein was expressed in EnBase® (fed-batch) cultivation mode. The pETiteTM N-His SUMO Kan vector, including anti-PcrV scFv gene, was transformed into Escherichia coli (BL21) cells. The expression and solubility of anti-PcrV scFv protein were investigated at two different temperatures (25 °C and 30 °C) and at different induction times (4, 6, 8, 12, and 24 hours). Results:Increased efficiency was achieved by EnBase® compared to LB broth; owing to the slow release of glucose, the maximum level of solubility and total protein expression was observed in EnBase® cultivation system at 30 °C and 24 h post induction. Furthermore, IC₅₀ for anti-PcrV scFv protein was determined to be approximately 7 μg/mL. Conclusion:Anti-PcrV scFv produced in this study showed promising in vitro results, protecting RBC from lysis by P. aeruginosa (exoU+). Key Words: Fed Batch, recombinant protein, Pseudomonas aeruginosa, scFv     

Simultaneous Extraction and Fractionation of Fish Oil from Tuna By-Product Using Supercritical Carbon Dioxide (SC-CO2)

ABSTRACT

Fish oil was extracted and simultaneously collected into six fractions based on molecular weight and the chain length of triglycerides in terms of fatty acid constituents without splitting of the triglycerides, using supercritical carbon dioxide (SC-CO₂) at optimized conditions of 40 MPa, 65°C, and a flow rate 3 mL min^?1. In each type of fractionation, the first fraction (F1) was rich in saturated fatty acids (SFA; 52.57 to 61.26%), followed by monounsaturated fatty acids (MUFA; 22.17 to 23.22%) and polyunsaturated fatty acids (PUFA; (0.54 to 20.37%); the sixth fraction (F6) was rich in PUFA (48.93%), followed by MUFA (33.59%) and SFA (13.61%). It was obvious that short-chain fatty acids were extracted at an earlier fraction; therefore, the latter fractions were dominant in long-chain fatty acids, especially MUFA and PUFA. Thus, omega-3 fish oil (last three fractions) was successfully separated to be used as a value-added health product.     

Efficacy of starter N fertilizer and rhizobia inoculant in dry pea (Pisum sativum Linn.) production in a semi-arid temperate environment

ABSTRACT

Uncertainties exist about the importance of rhizobia inoculant and starter nitrogen (N) application in dry pea (Pisum sativum L.) production. Three field experiments were conducted to evaluate how rhizobia inoculant and starter N fertilizer affect pea seed yield and protein concentration in a semi-arid environment in central Montana. Commercial rhizobia inoculant was mixed with seed prior to planting at the manufacturer’s recommended rate. Starter N fertilizers were applied into the same furrow as seed at 0, 22, 44 and 88 kg ha^?1 as urea, slow-release polymer-coated N fertilizer (ESN), and a combination of both. The application of rhizobia inoculant had no or a very small beneficial effect on pea yield in lands with a previous history of peas. In a land without pea history, application of rhizobia increased pea seed yield by 16%. The positive effect of starter N was only pronounced when initial soil N was low (≤ 10 kg ha^?1 nitrate-nitrogen), which increased net return by up to US$ 42 ha^?1. In this condition, application of slow-release N outperformed urea. However, application of starter N (especially with urea) had a negative effect on pea establishment, vigor and seed yield when soil initial N was high (≥ 44 kg ha^?1 NO₃-N). The results indicate that the rate, placement and form of the starter N must be optimized to benefit pea yield and protein without detrimental effects on germination and nodulation. Moreover, application of starter N must be guided by the soil nitrate content.     

Minimizing Ammonia Volatilization from Urea in Waterlogged Condition Using Chicken Litter Biochar

Application of urea in lowland rice fields leads to ammonia (NH₃) volatilization and environmental pollution, and diminishes nitrogen recovery by rice (Oryza sativa L.). Amending urea with biochar could reduce NH₃ loss from urea as well as improve chemical properties of acid soils. An incubation study was conducted using a closed-dynamic air flow system to determine NH₃ volatilization from urea and chemical properties of an acid soil (Typic Paleudults). The soil was mixed with three rates of chicken litter biochar (20, 40, and 60 g pot^?1) and 1.31 g urea. Mixing an acid soil with biochar (60 g pot^?1) in waterlogged to stimulate conditions in paddy condition significantly reduced NH₃ loss and total titratable acidity. Biochar application also increased soil pH, total nitrogen, available nitrate, organic matter, total organic carbon, total carbon, available phosphorus, and exchangeable cations. Thus, chicken litter biochar can be used to reduce urea-N loss and ameliorate chemical properties of acid soils. This aspect is being embarked on in our on-going field experiments.     

Comparative study of serological methods for the detection of antibodies to porcine reproductive and respiratory syndrome virus.

A comparison was made of serological diagnostic methods used for the detection of antibodies against porcine reproductive and respiratory syndrome (PRRS) virus. In the "phase I" PRRS test panel comparison, a panel of sera collected from 135 pigs of various ages, from North American herds with and without PRRS histories, were sent to 4 different laboratories and tested by an indirect immunofluorescent assay (IFA), an immunoperoxidase monolayer assay (IPMA) and an indirect enzyme-linked immunosorbent assay (iELISA). In the "phase II" PRRS test panel comparison, a panel of 382 sera collected from pigs of various ages, PRRS histories, and from various locations in North America and France, were divided into 2 panels (A & B) and sent to 3 Canadian laboratories and tested by the IFA and iELISA. In the phase I comparison, agreement between the IFA of laboratory 4 and the iELISA and IPMA of laboratory 3 was excellent (kappa values of 95% and 98%, respectively). This contrasted with the poor agreement between these laboratories and the IFA results of laboratories 1 and 2 in the phase I trial. In the phase II comparison, the results demonstrated good agreement between various tests both within and between laboratories. The overall performance of the iELISA was superior in the combination of sensitivity (96.1%) and specificity (100%) relative to the reference classification of the serum samples and repeatability (kappa value 98%). The iELISA is technically superior to IFA and IPMA, time efficient, cost effective and suitable for testing of a large number of samples over a short period of time. Thus, the iELISA may be a better alternative to IFA or IPMA for routine detection of PRRS viral antibodies in swine sera.     

Comparison of competitive ELISA, indirect ELISA and standard AGID tests for detecting blue-tongue virus antibodies in cattle and sheep

The performances of a competitive enzyme-linked immunosorbent assay (ELISA) using a group specific monoclonal antibody against bluetongue virus, an indirect ELISA and the standard agar gel immunodiffusion (AGID) test were compared in the detection of serum antibody against bluetongue virus. Test sera consisted of 1300 bovine, 530 ovine and 160 carpine samples from bluetongue-free areas of Canada, 605 bovine and ovine field samples from the USA and Barbados and 464 samples from 79 cattle and sheep experimentally infected with 19 South African and five USA serotypes of bluetongue virus. The diagnostic specificity of the competitive ELISA, as determined for the bluetongue virus-free cattle sera was superior (99.92 per cent) to that of the indirect ELISA (99.85 per cent) and the AGID (99.0 per cent). The specificities of the competitive ELISA for sheep (99.63 per cent) and goats (100.0 per cent) sera were also higher than those of the AGID test. The performance of the ELISA tests was similar whether a gamma-ray-irradiated (2.0 Mrad) or a non-irradiated bluetongue virus antigen preparation was used. The competitive ELISA results for bovine field sera from endemic areas demonstrated a relatively low level of agreement (92.04 per cent) with AGID test results, with 9.7 per cent false negatives. The possible presence in these sera of antibody to cross-reacting antigens or to other orbiviruses, eg, epizootic haemorrhagic disease virus, which react in the AGID but not in the competitive ELISA may account for this lack of agreement.(ABSTRACT TRUNCATED AT 250 WORDS)