Diagnostic real-time PCR assay for the quantitative detection of Theileria equi from equine blood samples

We developed a TaqMan real-time polymerase chain reaction (PCR) assay for the quantitative detection of Theileria equi from the in vitro-cultured parasite and field blood samples collected from horses living in Ghana and Brazil. The detection limit for the assay was determined to be 1.5 parasites/microl per sample, and the quantitative capacity was demonstrated using the in vitro-cultured parasite. For field applications, the real-time PCR assay was compared to a previously established nested PCR assay used as the gold standard for the real-time PCR assay. Of 65 field blood samples, 46 samples were T. equi-positive in the nested PCR assay, while the real-time PCR assay also detected the parasite in all 46 of the nested PCR-positive samples but did not detect T. equi in the remaining 19 negative blood samples. This quantitative real-time PCR assay provides a valuable tool for fast laboratory diagnostic assessment of T. equi infection in horses.     

Unrecognized diversity in New Guinean crayfish species (Decapoda,Parastacidae): The evidence from molecular data

The phylogenetic relationships among imported ornamental crayfish belonging to the genus Cherax were inferred from a combined dataset of 3 mitochondrial genes (COI, 16S and 12S) and by comparison with available GenBank sequences of 14 Cherax species. Furthermore, the concordance of previously described species obtained from a wholesaler (Cherax boesemani, C. holthuisi and C. peknyi) with available GenBank sequences was verified based on COI with special respect to comparison with sequences assigned as Cherax species. Recently described species C. gherardiae, C. pulcher and C. subterigneus belong to the northern group of Cherax species. Comparison and analysis with other GenBank COI sequences show previously unreported diversity of New Guinean species, suggesting 5 putative new species. Surprisingly, species assigned to the subgenus Astaconephrops do not form a monophyletic clade; this subgenus should be reappraised relative to the purported typical morphological characteristic of the uncalcified patch on male chelae. Increasing importation of crayfish underscores the importance of accurate species identification. Use of basic molecular methods is a necessary requisite for documenting occurrence, abundance and population trends of target species. Consequently, it helps to support eventual conservation decision‐making by stakeholders.     

Development of two immunochromatographic tests for the serodiagnosis of bovine babesiosis

In this study, we developed two immunochromatographic tests (ICTs), which are nitrocellulose membrane-based immunoassays for the convenient and rapid serodiagnosis of bovine babesiosis caused by Babesia bovis (BoICT) and Babesia bigemina (BiICT). The efficacy of two ICTs was evaluated using 13 positive sera from experimentally infected cattle with B. bovis or B. bigemina. Clear results showed that the BoICT and ELISA detected antibodies in sera collected from 14 to 93 days post-infection, while BiICT and ELISA detected from 13 to 274 days post-infection. In additon, non-infected cattle, Neospora caninum, and Cryptosporidium parvum were negative in two ICTs. To evaluate the field utility of the ICTs, we tested 186 field bovine sera collected from cattle living in Yanbian (China) and Mato Grosso do Sul (Brazil). The results of ICTs were compared to those of classical serodiagnostic methods, enzyme-linked immunosorbent assay (ELISA) and the indirect immunofluorescence assay (IFAT). The overall concordances of BoICT were determined as 92.5 and 90.3% when the results of ELISA and IFAT were set as the reference standards, respectively. In contrast, those of BiICT showed 96.8 and 92.5% relative to the results of standard ELISA and IFAT, respectively. Conventional and rapid diagnosic devices for bovine babesiosis may provide a valuable tool in clinical and field applications.     

The understudied and underappreciated role of predation in the mortality of fish released from fishing gears

The assumption that animals released from fishing gears survive has frequently been scrutinized by researchers in recent years. Mortality estimates from these research efforts can be incorporated into management models to ensure the sustainability of fisheries and the conservation of threatened species. Post‐release mortality estimates are typically made by holding the catch in a tank, pen or cage for short‐term monitoring (e.g. 48 h). These estimates may be inaccurate in some cases because they fail to integrate the challenges of the wild environment. Most obvious among these challenges is predator evasion. Stress and injury from a capture experience can temporarily impair physiological capacity and alter behaviour in released animals, a period during which predation risk is likely elevated. In large‐scale commercial fisheries, predators have adapted their behaviour to capitalize on impaired fishes being discarded, while in recreational catch‐and‐release fisheries, exercise and air exposure can similarly impede the capacity for released fish to evade opportunistic predators. Owing to the indirect and often cryptic nature of this source of mortality, very few studies have attempted to document it. A survey of the literature demonstrated that <2% of the papers in the combined realms of bycatch and catch‐and‐release have directly addressed or considered post‐release predation. Future research should combine field telemetry and laboratory studies using both natural and simulated predation encounters and incorporate physiological and behavioural endpoints. Quite simply, predation is an understudied and underappreciated contributor to the mortality of animals released from fishing gears.     

Interactions between physical and biotic factors influence CO2 flux in Antarctic dry valley soils

Soil carbon dioxide (CO₂) flux is an integrative measure of ecosystem functioning representing both biotic and physical controls over carbon (C) balance. In the McMurdo Dry Valleys of Antarctica, soil CO₂ fluxes (approximately −0.1-0.15 μmol m⁻² s⁻¹) are generally low, and negative fluxes (uptake of CO₂) are sometimes observed. A combination of biological respiration and physical mechanisms, driven by temperature and mediated by soil moisture and mineralogy, determine CO₂ flux and, therefore, soil organic C balance. The physical factors important to CO₂ flux are being altered with climate variability in many ecosystems including arid forms such as the Antarctic terrestrial ecosystems, making it critical to understand how climate factors interact with biotic drivers to control soil CO₂ fluxes and C balances. We measured soil CO₂ flux in experimental field manipulations, microcosm incubations and across natural environmental gradients of soil moisture to estimate biotic soil respiration and abiotic sources of CO₂ flux in soils over a range of physical and biotic conditions. We determined that temperature fluctuations were the most important factor influencing diel variation in CO₂ flux. Variation within these diel CO₂ cycles was explained by differences in soil moisture. Increased temperature (as opposed to temperature fluctuations) had little or no effect on CO₂ flux if moisture was not also increased. We conclude that CO₂ flux in dry valley soils is driven primarily by physical factors such as soil temperature and moisture, indicating that future climate change may alter the dry valley soil C cycle. Negative CO₂ fluxes in arid soils have recently been identified as potential net C sinks. We demonstrate the potential for arid polar soils to take up CO₂, driven largely by abiotic factors associated with climate change. The low levels of CO₂ absorption into soils we observed may not constitute a significant sink of atmospheric CO₂, but will influence the interpretation of CO₂ flux for the dry valley soil C cycle and possibly other arid environments where biotic controls over C cycling are secondary to physical drivers.     

Estimating nocturnal ecosystem respiration from the vertical turbulent flux and change in storage of CO2

Micrometeorological measurements of nighttime ecosystem respiration can be systematically biased when stable atmospheric conditions lead to drainage flows associated with decoupling of air flow above and within plant canopies. The associated horizontal and vertical advective fluxes cannot be measured using instrumentation on the single towers typically used at micrometeorological sites. A common approach to minimize bias is to use a threshold in friction velocity, u_*, to exclude periods when advection is assumed to be important, but this is problematic in situations when in-canopy flows are decoupled from the flow above. Using data from 25 flux stations in a wide variety of forest ecosystems globally, we examine the generality of a novel approach to estimating nocturnal respiration developed by van Gorsel et al. (van Gorsel, E., Leuning, R., Cleugh, H.A., Keith, H., Suni, T., 2007. Nocturnal carbon efflux: reconciliation of eddy covariance and chamber measurements using an alternative to the u_*-threshold filtering technique. Tellus 59B, 397–403, Tellus, 59B, 307-403). The approach is based on the assumption that advection is small relative to the vertical turbulent flux (F_C) and change in storage (F_S) of CO₂ in the few hours after sundown. The sum of F_C and F_S reach a maximum during this period which is used to derive a temperature response function for ecosystem respiration. Measured hourly soil temperatures are then used with this function to estimate respiration R_Rmax. The new approach yielded excellent agreement with (1) independent measurements using respiration chambers, (2) with estimates using ecosystem light-response curves of F_c + F_s extrapolated to zero light, R_LRC, and (3) with a detailed process-based forest ecosystem model, R_cast. At most sites respiration rates estimated using the u_*-filter, R_ust, were smaller than R_Rmax and R_LRC. Agreement of our approach with independent measurements indicates that R_Rmax provides an excellent estimate of nighttime ecosystem respiration.     

Voluntary institutions and behaviours as alternatives to formal regulations in recreational fisheries management

Traditional regulatory options (formal institutions) imposed by government agencies such as harvest and gear restrictions represent the standard in recreational fisheries management, at least in developed countries. However, there exist a number of alternatives including the use of angler education programmes that attempt to evoke voluntary changes in angler behaviour, resulting in the emergence of voluntarily motivated resource‐conserving informal institutions. These ‘softer’ approaches to aquatic stewardship and fisheries management can be developed in cooperation with stakeholders and in many cases are led by avid anglers and angling groups. Examples of such measures include voluntary sanctuaries, informally enforced seasonal closures, personal daily bag limits, self‐imposed constraints on gear, development of entirely live‐release fisheries, and adoption of fish and aquatic ecosystem conservation‐oriented gears and release practices. Education efforts that provide anglers with knowledge on best practices and empower them to modify their behaviour hold great promise to meet formal management goals and objectives, but seem to be underutilized relative to formal regulations. This article highlights the benefits and challenges of relying on informal institutions as alternatives to traditional regulatory options. Informal institutions that protect resources and help overfished stocks recover hold great promise in both developed and developing countries, particularly when there is a single stakeholder group or when the capacity to enforce traditional regulations or to invest in stock assessments is limited. Informal institutions may help make formal institutions more effective or can even be alternatives to costly institutions that depend on enforcement to be effective.     

Antioxidant Effect of Dietary Supplement Withania somnifera L. Reduce Blood Glucose Levels in Alloxan-Induced Diabetic Rats

The phenolic compounds and flavonoids were determined from the extracts of Withania somnifera root (WSREt) and leaf (WSLEt). The WSREt has 28.26 mg/g total phenolic compounds and 17.32 mg/g flavonoids, whereas WSLEt has 5.4 mg/g total phenolic compounds and 5.1 mg/g flavonoids. The WSREt, WSLEt and glibenclamide were orally administered daily to diabetic rats for 8 weeks. After the treatment, the levels of urine sugar, blood glucose, liver glycogen, and antioxidants like vitamin C and E in plasma and superoxide dismutase (SOD), catalase (CAT), thiobarbituric acid reactive substances (TBARS), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and reduced glutathione (GSH) in liver, kidney and heart were determined. Diabetic rats showed a significant (p < 0.05) elevation in glucose and TBARS and a significant (p < 0.05) reduction in glycogen, vitamin C and E, SOD, CAT, GPx, GST, and GSH levels when compared to normal control rats. Administration of WSREt, WSLEt and glibenclamide to diabetic rats restored the levels to normal. In the light of aforesaid facts, it is suggested that the presence of phenolic compounds including flavonoids in W. somnifera root and leaf extracts and their antioxidant activity may play a vital role in reduction of blood glucose level in alloxan-induced diabetic rats.     

All three classes of CpG ODNs up-regulate IP-10 gene in pigs

The analysis of CpG ODN induced innate immune responses in different animal species has shown substantial similarities and differences in levels and types of induced cytokines profile. The objectives of these studies were to identify innate immune biomarkers activated by three classes of CpG ODNs in pigs. For this purpose, we investigated the kinetics of innate immune responses in immune cells from pigs following in vitro and in vivo stimulation with CpG ODNs. The mRNA expression of cytokine and chemokine genes were assayed by SYBR^@ green based quantitative real time PCR. A-class CpG ODN induced significant but transient levels of IFN-γ, IL-12 (P40), IL-6, IL-4 and TNF-α mRNA, C-class CpG ODN induced significant level of IFN-γ, IFN-α and IL-12 mRNA and the lowest level of IL-4 (Th-2 type) mRNA. A very low level of some cytokines stimulation was observed by GC ODNs. It is noteworthy, that IL-12 (P35) mRNA was significantly stimulated by B-class GpC ODN 7909. Interestingly, all classes of CpG ODNs induced significant level of IP-10 at 12 h post stimulation. These in vitro and in vivo observations suggest that interferon-γ inducible protein 10 (IP-10) may be a reliable biomarker for immune activity induced by CpG ODNs in pigs.