Evaluation of an electrolyte analyzer for measurement of ionized calcium and magnesium concentrations in blood, plasma, and serum of dogs

OBJECTIVE: To evaluate an electrolyte analyzer for measurement of ionized calcium (Ca(i)) and magnesium (Mg(i)) concentrations in blood, plasma, and serum; investigate the effect of various factors on measured values; and establish reference ranges for Ca(i) and Mg(i) in dogs. ANIMALS: 30 healthy adult dogs of various breeds. PROCEDURE: Precision in a measurement series, day-to-day precision, and linearity were used to evaluate the analyzer. The effects of exposure of serum samples to air, type of specimen (blood, plasma, or serum), and storage temperature on sample stability were assessed. Reference ranges were established with anaerobically handled serum. RESULTS: The coefficient of variation for precision in a measurement series was < or = 1.5% for both electrolytes at various concentrations. The Ca(i) and Mg(i) concentrations were significantly lower in aerobically handled serum samples, compared with anaerobically handled samples. The Ca(i) and Mg(i) concentrations differed significantly among blood, plasma, and serum samples. In anaerobically handled serum, Ca(i) was stable for 24 hours at 22 degrees C, 48 hours at 4 degrees C, and 11 weeks at -20 degrees C; Mg(i) was stable for 8 hours at 22 degrees C, < 24 hours at 4 degrees C, and < 1 week at -20 degrees C. In anaerobically handled serum, reference ranges were 1.20 to 1.35 mmol/L for Ca(i) and 0.42 to 0.58 mmol/L for Mg(i). CONCLUSIONS AND CLINICAL RELEVANCE: The electrolyte analyzer was suitable for determination of Ca(i) and Mg(i) concentrations in dogs. Accurate results were obtained in anaerobically handled serum samples analyzed within 8 hours and kept at 22 degrees C.     

Ocular and multicentric lymphoma in a young racehorse

A 6-year-old thoroughbred gelding was presented with a history of blepharospasm and opacity in the OS of 1 weeks' duration. Ophthalmic examination findings were consistent with acute uveitis in the OS, and traditional treatment was initiated with systemic antibiotics and anti-inflammatory drugs, topical mydriatics, and corticosteroids. During the total treatment period of 4 weeks response to treatment was weak and the horse developed further problems such as cellulitis of the right hind limb with fever and eventually weight loss and dependent edema. Blood work was indicative of liver disease. Abdominal sonography revealed severe splenomegaly and slight hepatomegaly, and a liver biopsy confirmed malignant T-cell lymphoma. The horse was euthanized due to deteriorating general condition and subsequently underwent postmortem examination. Necropsy and histologic examination revealed a multicentric lymphoma with involvement of spleen, mesenteric lymph nodes, and OU.
The findings in this case demonstrate that the differential diagnosis of intraocular and systemic lymphoma should be considered in any horse presenting with anterior uveitis, especially when uveitis is unresponsive to treatment and when additional systemic signs of illness such as lethargy, fever, weight loss, or dependent edema arise.
Cytological examination of aqueous humor may provide a rapid diagnosis of intraocular lymphoma in eyes with clinical uveitis.     

Diagnosis of canine Hepatozoon spp. infection by quantitative PCR

Hepatozoon (H.) americanum and H. canis are the etiological agents of canine hepatozoonosis, a disease that is found worldwide and is also prevalent in the southeastern United States. Current laboratory diagnosis of canine hepatozoonosis caused by H. americanum is usually dependent on visual identification of Hepatozoon "onion skin cysts" in muscle biopsies, an approach that requires invasive sampling and can result in false negatives. We have developed a diagnostic method for detection of Hepatozoon spp. DNA that integrates nucleic acid extraction with extensive agitation to maximize DNA extraction efficiency. The DNA extracted from canine EDTA-whole blood is subjected to real-time PCR, and fluorescence resonance energy transfer (FRET) probes detect a signature polymorphism in the amplified DNA. This PCR method amplifies a fragment of the Hepatozoon 18S rDNA gene, detects as few as 7 genomic copies of Hepatozoon spp. per ml of blood with high specificity, and differentiates between H. americanum and H. canis amplicons. A surprising 300-fold increase of H. americanum 18S rDNA targets occurred during 3-0 days of storage of positive blood specimens. Examination of 614 EDTA-blood samples submitted mostly from the southeastern Unites States from dogs with suspected hepatozoonosis identified H. americanum in 167 samples (27.2%). An additional 14 samples (2.3%) were positive for H. canis, and 14 samples (2.3%) were positive for both H. americanum and H. canis. These results suggest that the Hepatozoon spp. 18S rDNA quantitative PCR may be a valuable tool that can improve diagnosis and therapy of canine hepatozoonosis.     

Parachlamydia acanthamoebae in domestic cats with and without corneal disease

Corneal samples of cats with and without corneal diseases were screened with a pan‐Chlamydiales PCR and specific PCRs for Parachlamydia, Protochlamydia, Chlamydophila felis, Acanthamoeba and feline herpesviruses (FHV‐1). Several corneal samples tested positive for Parachlamydia and related Chlamydiales, indicating cat exposure to these intracellular bacteria.     

Combining hydrological modeling and GIS approaches to determine the spatial distribution of groundwater recharge in an arid irrigation scheme

Accurate quantification of the rate of groundwater (GW) recharge, a pre-requisite for the sustainable management of GW resources, needs to capture complex processes, such as the upward flow of water under shallow GW conditions, which are often disregarded when estimating recharge at a larger scale. This paper provides (1) a method to determine GW recharge at the field level, (2) a consequent procedure for up-scaling these findings from field to irrigation scheme level and (3) an assessment of the impacts of improved irrigation efficiency on the rate of GW recharge. The study is based on field data from the 2007 growing season in a Water Users Association (WUA Shomakhulum) in Khorezm district of Uzbekistan, Central Asia, an arid region that is characterized by a predominance of cotton, wheat and rice under irrigation. Previous qualitative studies in the region reported irrigation water supplies far above the crop water requirements, which cause GW recharge. A field water balance model was adapted to the local irrigation scheme; recharge was considered to be a fraction of the irrigation water losses, determined as the difference between net and gross irrigation requirements. Capillary rise contribution from shallow GW levels was determined with the HYDRUS-1D model. Six hydrological response units (HRUs) were created based on GW levels and soil texture using GIS and remote sensing techniques. Recharge calculated at the field level was up-scaled first to these HRUs and then to the whole WUA. To quantify the impact of improved irrigation efficiency on recharge rates, four improved irrigation efficiency scenarios were developed. The area under cotton had the second highest recharge (895 mm) in the peak irrigation period, after rice with 2,514 mm. But with a low area share of rice in the WUA of <1 %, rice impacted the total recharge only marginally. Due to the higher recharge rates of cotton, which is grown on about 40 % of the cropped area, HRUs with a higher share of cotton showed higher recharge (9.6 mm day^?1 during August) than those with a lower share of cotton (4.4 mm day^?1). The high recharge rates in the cotton fields were caused by its water requirements and the special treatment given to this crop by water management planners due to its strategic importance in the country. The scenario simulations showed that seasonal recharge under improved irrigation efficiency could potentially be reduced from 4 mm day^?1 (business-as-usual scenario) to 1.4 mm day^?1 (scenario with maximum achievable efficiency). The combination of field-level modeling/monitoring and GIS approaches improved recharge estimates because spatial variability was accounted for, which can assist water managers to assess the impact of improved irrigation efficiencies on groundwater recharge. This impact assessment enables managers to identify options for a recharge policy, which is an important component of integrated management of surface and groundwater resources.     

Biochar application to temperate soils: Effects on soil fertility and crop growth under greenhouse conditions

Biochar (BC) application as a soil amendment has aroused much interest and was found to considerably improve soil nutrient status and crop yields on poor, tropical soils. However, information on the effect of BC on temperate soils is still insufficient, with effects expected to differ from tropical soils. We investigated the effects of BC on soil nutrient dynamics, crop yield, and quality in a greenhouse pot experiment. We compared three agricultural soils (Planosol, Cambisol, Chernozem), and BCs of three different feedstocks (wheat straw [WS], mixed woodchips [WC], vineyard pruning [VP]) slowly pyrolyzed at 525°C, of which the latter was also pyrolyzed at 400°C. The BCs were applied at two rates (1% and 3%, which would correspond to 30 and 90 t ha^–1 in the field). Three crops, namely mustard (Sinapis alba L.), barley (Hordeum vulgare L.), and red clover (Trifolium pretense L.) were grown successively within one year. The investigated soil properties included pH, electrical conductivity (EC), cation‐exchange capacity (CEC), calcium‐acetate‐lactate (CAL)–extractable P (P_CAL) and K (K_CAL), C, N, and nitrogen‐supplying potential (NSP). The results show a pH increase in all soils. The CEC increased only on the Planosol. The C : N ratio increased at 3% application rate. Despite improving the soil nutrient status partly, yields of the first crop (mustard) and to a lesser extent of the second crop (barley) were significantly depressed through BC application (by up to 68%); the yield of clover as third crop was not affected. Only the BC from WS maintained yields in the range of the control and even increased barley yield by 6%. The initial yield reduction was accompanied by notable decreases (Cu, Fe, Mn, Zn) and increases (Mo) in micronutrient concentrations of plant tissues while nitrogen concentrations were hardly affected. The results of the pot experiment show that despite additional mineral fertilization, short‐term growth inhibition may occur when applying BC without further treatment to temperate soils.     

BiKF AdaMus: a novel research project studying the response and adaptive potential of single species and communities to climate change in combination with other stressors

Introduction

How will the combination of climate-induced physical/chemical changes and anthropogenic pollution impact key species and biodiversity, and thus the ecosystem functions in future?

Discussion

The project AdaMus within the newly founded Biodiversity and Climate Research Centre BiK^F aims to understand the prospective ecosystem stress responses and adaptive potential of species and communities to multiple stressors to provide reliable predictions of future developments in terrestric, aquatic and marine environments.

Conclusion

Long-term experiments with terrestrial, aquatic and marine species and communities will be conducted under IPCC predicted climate conditions. In terrestric and limnic experiments, additional co-stressors such as pesticides and alien species are introduced to test for interactions. In the case of marine plankton, potential consequences of multiple climate-related changes in the physical environment (temperature, salinity and oxygen) will be analysed.     

Type, density, and location of immune cells within human colorectal tumors predict clinical outcome

The role of the adaptive immune response in controlling the growth and recurrence of human tumors has been controversial. We characterized the tumor-infiltrating immune cells in large cohorts of human colorectal cancers by gene expression profiling and in situ immunohistochemical staining. Collectively, the immunological data (the type, density, and location of immune cells within the tumor samples) were found to be a better predictor of patient survival than the histopathological methods currently used to stage colorectal cancer. The results were validated in two additional patient populations. These data support the hypothesis that the adaptive immune response influences the behavior of human tumors. In situ analysis of tumor-infiltrating immune cells may therefore be a valuable prognostic tool in the treatment of colorectal cancer and possibly other malignancies.     

Cloudy apple juice is more effective than apple polyphenols and an apple juice derived cloud fraction in a rat model of colon carcinogenesis

As recently shown, a cloudy apple juice (CloA) was effective to modulate colon cancer associated parameters in rats treated with 1,2-dimethylhydrazine (DMH). To identify the bioactive substance classes in CloA, we fractionated CloA to yield a total polyphenol (PF) and a cloud (CF) fraction consisting of proteins, fatty acids, polyphenols, and cell wall polysaccharides. Rats received water (control (Cont)) or CloA, PF, and CF separate or combined (PF-CF) ad libitum for 7 weeks starting one week before the first DMH-injection. As determined by comet assay, the DMH-induced genotoxicity in colonocytes of controls (Cont/DMH: 7.7 +/- 0.5%) was significantly reduced by CloA (3.3 +/- 0.3%) but not by any of the fractions. The crypt cell proliferation induced by DMH (Cont/NaCl: 7.5 +/- 0.6%; Cont/DMH: 14.9 +/- 0.8%) was significantly decreased by CloA (9.4 +/- 0.4%), PF (12.4 +/- 0.7%), CF (11.6 +/- 0.4%), and PF-CF (12.4 +/- 0.6%). Although not statistically significant, CloA tended to reduce the number of large aberrant crypt foci (ACF) (Cont/DMH: 19.0 +/- 3.7; CloA/DMH: 12.3 +/- 1.9), while none of the fractions affected ACFs. Neither CloA nor the fractions changed mRNAs of colonic cyclooxygenases (COX-1, COX-2), glutathione-associated enzymes (GST-M2, gamma-GCS, GST-P), the splenocyte CD4/CD8 ratio, natural killer cell activity, and plasma antioxidant status. These results demonstrate that CloA had a higher cancer-preventive potential than the fractions and further, besides PF, identified CF as an additional bioactive fraction of CloA.