In vitro identification and verification of inflammatory biomarkers in swine

Currently there are no non-steroidal anti-inflammatory drugs (NSAIDs) approved for the control of inflammation in swine due to a lack of validated animal models and suitable biomarkers to assess drug efficacy. This study investigates the differential expression of genes altered in response to Escherichia coli lipopolysaccharide (LPS) induced inflammation which may serve as indicators of NSAID efficacy. Unstimulated whole blood from swine was mixed with tissue culture media, stimulated with LPS, and RNA extracted at the following time points 0 h, 1 h, 3 h, 24 h and 48 h. Total RNA was extracted and analyzed using a commercial swine DNA microarray. The DNA microarray was utilized as a screen to determine potential biomarkers, focusing on the genes that exhibited the greatest degree of differential expression. A master list of 57 genes was formed based on the differential expression as a result of the stimulation. Following analysis, 12 genes whose expressions were significantly altered (8 up- and 4 down-regulated) were chosen for verification via quantitative RT-PCR (qRT-PCR). The qRT-PCR analysis confirmed the differential expression of 11 of the 12 genes chosen via the microarray analyses. Specifically, traditional genes such as SAA, G-CSF, and IL-10 were up-regulated, while CD4 was down-regulated; all of the genes were altered by 24 h or 48 h post-stimulation. We demonstrate here that expression of these 11 genes is altered as a direct result of LPS stimulation and consequently inflammation.     

Associations between Theileria orientalis Ikeda type infection and the growth rates and haematocrit of suckled beef calves in the North Island of New Zealand

AIMS: The principle aim of this study was to examine the association between infection with Theileria orientalis Ikeda type and growth rates of suckled beef calves on four beef farms. In addition, associations between calf sex, sampling time, and individual farm and T. orientalis Ikeda type infection intensity and haematocrit (HCT) were investigated.

METHODS: The study was a prospective longitudinal study in which 240 calves from four purposively selected beef farms in the North Island of New Zealand were blood sampled and weighed in late spring, mid-summer and early autumn. Two farms were from high-risk (A and B) and two from low-risk (C and D) tick areas. Blood samples were analysed to determine HCT, and the number of T. orientalis Ikeda type organisms/µL of blood (infection intensity) using a quantitative PCR assay. A calf was defined as infected if >415 organisms/µL were detected in a blood sample. Linear mixed models were used to examine associations between infection intensity, mean daily liveweight gain (MDG), HCT, calf sex and time of sampling on the four farms.

RESULTS: On Farms A and B nearly all calves were infected at each sampling time, on Farm C <30% were infected at any sampling and on Farm D infection prevalence increased from 32 to 79% between late spring and early autumn. On Farms C and D, from mid-summer to early autumn, mean MDG was 0.127 (95% CI=0.072–0.183) kg/day less for infected than uninfected calves (p<0.001). On all farms MDG was negatively associated with infection intensity for mid-summer and early autumn sampling times (p=0.037). The relationship between time of sampling and infection intensity varied between farms (p<0.001), and between male and female calves (p=0.018). Females had a higher infection intensity than males at the mid-summer and early autumn samplings. The association between HCT and infection intensity varied with sampling time and farm (p=0.018). There was a strong negative association between infection intensity and HCT at the late spring sampling, but in mid-summer there was no association, and in early autumn only a weak association.

CONCLUSIONS AND CLINICAL RELEVANCE: This study has shown that beef farmers in the North Island of New Zealand should be concerned about the welfare effects and economic impacts of T. orientalis Ikeda type infection in suckled beef calves.     

Importance of oil overlay for production of porcine embryos in vitro

Technologies to edit the zygote genome have revolutionized biomedical research not only for the creation of animal models for the study of human disease but also for the generation of functional human cells and tissues through interspecies blastocyst complementation technology. The pig is the ideal species for these purposes due to its great similarity in anatomy and physiology to humans. Emerging biotechnologies require the use of oocytes and/or embryos of good quality, which might be obtained using in vitro production (IVP) techniques. However, the current porcine embryo IVP systems are still suboptimal and result in low monospermic fertilization and blastocyst formation rates and poor embryo quality. During recent years, intensive investigations have been performed to evaluate the influence of specific compounds on gametes and embryos and to avoid the use of undefined supplements (serum and serum derivate) in the incubation media. However, little consideration has been given to the use of the mineral oil (MO) to overlay incubation droplets, which, albeit being a routine component of the IVP systems, is a totally undefined and thus problematic product for the safety of gametes and embryos. In this review, we provide an overview on the advantages and disadvantages of using MO to cover the incubation media. We also review one important concern in IVP laboratories: the use of oils containing undetected contamination. Finally, we discuss the effects of different types of oils on the in vitro embryo production outcomes and the transfer of compounds from oil into the culture media.     

Vektorenbekämpfung bei Viruskrankheiten im Gemüsebau?

Ohne Zusammenfassung     

Changes in total, mineralizable and light fraction soil organic matter with cropping and tillage intensities in semiarid southern Alberta, Canada

There has been a trend toward increased cropping intensity and decreased tillage intensity in the semiarid region of the Canadian prairies. The impact of these changes on sequestration of atmospheric CO₂ in soil organic carbon (C) is uncertain. Our objective was to quantify the changes in total, mineralizable and light fraction organic C and nitrogen (N) due to the adoption of continuous cropping and conservation tillage practices. We sampled three individual long-term experiments at Lethbridge, Alberta, in September 1992: a spring wheat (Triticum aestivum L.)-fallow tillage study, a continuous spring wheat tillage study and a winter wheat rotation-tillage study. Treatments had been in place for 3–16 years. In the spring wheat-fallow study, different intensities (one-way disc > heavy-duty cultivator > blade cultivator) of conventional tillage (CT) were compared with minimum tillage (MT) and zero tillage (ZT). After 16 years, total organic C was 2.2 Mg ha⁻¹ lower in more intensively worked CT treatments (one-way disc, heavy-duty cultivator) than in the least-intensive CT treatment (blade cultivator). The CT with the blade cultivator and ZT treatments had similar levels of organic C. The CT treatments with the one-way disc and heavy-duty cultivator had light fraction C and N and mineralizable N amounts that were about 13–18% lower than the CT with the blade cultivator, MT or ZT treatments. In the continuous spring wheat study, 8 years of ZT increased total organic C by 2 Mg ha⁻¹, and increased mineralizable and light fraction C and N by 15–27%, compared with CT with a heavy-duty cultivator prior to planting. In the winter wheat rotation-tillage study, total organic C was 2 Mg ha⁻¹ higher in a continuous winter wheat (WW) rotation compared with that in a winter wheat-fallow rotation. The lack of an organic C response to ZT on the WW rotation may have been due to moldboard plowing of the ZT treatment in 1989 (6 years after establishment and 3 years before soil sampling), in an effort to control a severe infestation of downy brome (Bromus tectorum L.). Our results suggest that although relative increases in soil organic matter were small, increases due to adoption of ZT were greater and occurred much faster in continuously cropped than in fallow-based rotations. Hence intensification of cropping practices, by elimination of fallow and moving toward continuous cropping, is the first step toward increased C sequestration. Reducing tillage intensity, by the adoption of ZT, enhances the cropping intensity effect.     

Evaluation of the use of sulfur dioxide to reduce postharvest losses on dark and green figs

Postharvest diseases limit the storage period and market life of fresh figs (Ficus carica L.). The objective of this work was to determine the effect of sulfur dioxide (SO₂) applied by fumigation and/or by dual release SO₂ generating pads on postharvest decay and quality retention of ‘Black Mission’ and ‘Brown Turkey’ (dark skin), and ‘Kadota’ and ‘Sierra’ (green skin) figs. A protocol for the computer-controlled application of gaseous SO₂ has been developed which allows the application of very low specific concentration × time products of SO₂ and simultaneous monitoring of the application progress. In vitro tests with important fungal, yeast and bacterial postharvest pathogens plated on Petri dishes and exposed to a SO₂ concentration × time product (C × t) of 100 (μL/L) h at different temperatures showed fewer survived at 20 °C than at 0 °C. Therefore, fumigations were carried out at 20 °C in the rest of the experiments. The evaluation of different SO₂ concentration × time products showed that a product of 25 (μL/L) h provided the best compromise between decay control and fruit injury. The performance of SO₂ fumigations on warm or cold fruit, its combination with SO₂ generating pads, and the use of repeated fumigations during cold storage were also evaluated. All the SO₂ treatments tested reduced the percentage of decay, extending the market life of fresh figs. However, in some cases, the use of SO₂ generating pads increased the incidence of skin bleaching. Fumigation of warm fruit at 25 (μL/L) h of SO₂ reduced populations of Alternaria and Rhizopus spp. growing on the fig surface. The treatment was more effective against Rhizopus spp. than against Alternaria spp. Contamination of fruit by Botrytis spp. and Penicillium spp. was also reduced by SO₂. In conclusion, results showed that SO₂ can be a potential tool to control postharvest rots and therefore increase the market life of fresh figs.     

Predatory dinosaur remains from madagascar: implications for the cretaceous biogeography of gondwana

Recent discoveries of fossil vertebrates from the Late Cretaceous of Madagascar include several specimens of a large theropod dinosaur. One specimen includes a nearly complete and exquisitely preserved skull with thickened pneumatic nasals, a median frontal horn, and a dorsal projection on the parietals. The new materials are assigned to the enigmatic theropod group Abelisauridae on the basis of a number of unique features. Fossil remains attributable to abelisaurids are restricted to three Gondwanan landmasses: South America, Madagascar, and the Indian subcontinent. This distribution is consistent with a revised paleogeographic reconstruction that posits prolonged links between these landmasses (via Antarctica), perhaps until late in the Late Cretaceous.