Evaluation of reticulocyte hemoglobin content (RET‐He) in the diagnosis of iron‐deficient erythropoiesis in dogs

Background

Reticulocyte hemoglobin content provided by the Siemens ADVIA (CHr) is an established marker of iron deficiency. The IDEXX ProCyte Dx hematology analyzer now provides a similar variable, reticulocyte hemoglobin equivalent (RET‐He).

Objectives

The objective was to evaluate RET‐He and its diagnostic utility in dogs, and to calculate a cutoff value for diagnosing iron‐deficient erythropoiesis (IDE). Furthermore, the prevalence of RET‐He values below this cutoff value was established.

Methods

One hundred and seventy‐one CBCs of healthy dogs were used to establish a RI. Stability of RET‐He was evaluated by repeated measurements over 48 hours (n = 10). The 25‐run coefficient of variation (CV) was calculated, and correlation and bias between measurements of RET‐He and CHr were assessed (n = 190). A cutoff value for diagnosing IDE was calculated. The utility of RET‐He in the detection of IDE was evaluated in 123 dogs. The prevalence of low RET‐He values was assessed retrospectively in a multicenter study (2012–2014) under participation of 7 veterinary clinics.

Results

Reticulocyte hemoglobin equivalent with an RI of 22.2 to 28.6 pg was statistically stable over 48 hours (P = .10). The CV was 1.8%. A fair correlation (ρ = 0.74) between RET‐He and CHr with a small bias of ?0.6 pg was found. The cutoff value for diagnosing IDE was 20.9 pg (sensitivity: 85%; specificity: 99%). The prevalence of RET‐He values below 20.9 pg was 10.3% (1084/10,553 dogs).

Conclusions

RET‐He on the ProCyte Dx is a precise screening tool in dogs to detect iron‐deficient erythropoiesis.     

Negligible contribution from roots to soil-borne phospholipid fatty acid fungal biomarkers 18:2ω6,9 and 18:1ω9

The phospholipid fatty acid biomarkers 18:1ω9, 18:2ω6,9 and 18:3ω3,6,9 are commonly used as fungal biomarkers in soils. They have, however, also been found to occur in plant tissues, such as roots. Thus, the use of these PLFAs as fungal biomarkers in sieved soil, which may still contain small remains of roots, has been questioned. We used data from a recent beech tree girdling experiment to calculate the contribution of roots to these biomarkers and were able to demonstrate that not more than 0.61% of 18:1ω9 and 18:2ω6,9 in sieved soil samples originated from roots (but 4% of 18:3ω3,6,9). Additionally, the abundance of the biomarker 18:2ω6,9 in the soil was found to be highly correlated to ectomycorrhizal root colonization, which further corroborates its fungal origin. PLFA biomarkers were substantially reduced in vital roots from girdled trees compared to roots of control trees (by up to 76%), indicating that the major part of PLFAs measured in roots may actually originate from ectomycorrhizal fungi growing inside the roots. We calculated, that even a near to 50% reduction in fine root biomass - as observed in the girdling treatment - accounted for only 0.8% of the measured decrease of 18:2ω6,9. Our results demonstrate that both 18:1ω9 and 18:2ω6,9 are suitable biomarkers for detecting fungal dynamics in soils and that especially 18:2ω6,9 is a reliable biomarker to study mycorrhizal dynamics in beech forests.     

Einfluß der Probenvorbereitung auf die Adsorption von Cd und Cu in sauren Waldböden

Influence of Soil Sample Preparation on Cd and Cu Adsorption in Acid Forest Soils The influence of soil sample preparation on Cd and Cu adsorption was investigated using six acid forest soil samples and comparing adsorption isotherms for fresh and air-dried samples. While no influence of sample preparation on Cd adsorption capacity was found, air-drying resulted in a significant decrease in Cu adsorption density in all six soil samples under investigation at ecotoxicologically relevant concentrations in the soil solution.     

A parametric and microstructural study of the formation of gluten network in mixed flour–water batter

The mechanism of gluten network development is still unclear and remains difficult to study since gluten network formation in bread dough is a rather quick process. In order to better characterize this dynamic event, we slowed down its kinetics by increasing the dough water content. During mixing, performed with a planetary mixer at variable mixing speeds and flour/water ratios, the torque was recorded. Common flours from wheat cultivars Orvantis, Caphorn and Isengrain, similar in composition and Farinograph parameters, were studied.     

Leaching of carbon and nitrogen from a sandy soil substrate: A comparison between suction plates and ion exchange resins

To determine boundary effects on leaching, we investigated (1) how filter materials affect the concentrations of dissolved organic carbon (DOC) and nitrate (NO₃‐N) in soil percolates and (2) whether ion exchange resins and suction plates are equally suited to capture NO₃‐N. DOC leaching was higher with PE suction plates and plate material did not affect NO₃‐N leachate concentrations. Cumulative NO₃‐N leaching was similar for glass suction plates and ion exchange resins.     

Microbial use of organic amendments in saline soils monitored by changes in the C/C ratio

An incubation experiment was carried out to investigate whether salinity at high pH has negative effects on microbial substrate use, i.e. the mineralization of the amendment to CO₂ and inorganic N and the incorporation of amendment C into microbial biomass C. In order to exploit natural differences in the ¹³C/¹²C ratio, substrate from two C₄ plants, i.e. highly decomposed and N-rich sugarcane filter cake and less decomposed N-poor maize leaf straw, were added to two alkaline Pakistani soils differing in salinity, which had previously been cultivated with C₃ plants. In soil 1, the additional CO₂ evolution was equivalent to 65% of the added amount in the maize straw treatment and to 35% in the filter cake treatment. In the more saline soil 2, the respective figures were 56% and 32%. The maize straw amendment led to an identical immobilization of approximately 48 μg N g⁻¹ soil over the 56-day incubation in both soils compared with the control soils. In the filter cake treatment, the amount of inorganic N immobilized was 8.5 μg N g⁻¹ higher in soil 1 than in soil 2 compared with the control soils. In the control treatment, the content of microbial biomass C₃-C in soil 1 was twice that in soil 2 throughout the incubation. This fraction declined by about 30% during the incubation in both soils. The two amendments replaced initially similar absolute amounts of the autochthonous microbial biomass C, i.e. 50% of the original microbial biomass C in soil 1 and almost 90% in soil 2. The highest contents of microbial biomass C₄-C were equivalent to 7% (filter cake) and 11% (maize straw) of the added C. In soil 2, the corresponding values were 14% lower. Increasing salinity had no direct negative effects on microbial substrate use in the present two soils. Consequently, the differences in soil microbial biomass contents are most likely caused indirectly by salinity-induced reduction in plant growth rather than directly by negative effects of salinity on soil microorganisms.     

Detection of Bartonella spp. in wild rodents in Israel using HRM real-time PCR

The prevalence of Bartonella spp. in wild rodents was studied in 19 geographical locations in Israel. One hundred and twelve rodents belonging to five species (Mus musculus, Rattus rattus, Microtus socialis, Acomys cahirinus and Apodemus sylvaticus) were included in the survey. In addition, 156 ectoparasites were collected from the rodents. Spleen sample from each rodent and the ectoparasites were examined for the presence of Bartonella DNA using high resolution melt (HRM) real-time PCR. The method was designed for the simultaneous detection and differentiation of eight Bartonella spp. according to the nucleotide variation in each of two gene fragments (rpoB and gltA) and the 16S–23S intergenic spacer (ITS) locus, using the same PCR protocol which allowed the simultaneous amplification of the three different loci. Bartonella DNA was detected in spleen samples of 19 out of 79 (24%) black rats (R. rattus) and in 1 of 4 (25%) Cairo spiny mice (A. cahirinus). In addition, 15 of 34 (44%) flea pools harbored Bartonella DNA. Only rat flea (Xenopsyla cheopis) pools collected from black rats (R. rattus) were positive for Bartonella DNA. The Bartonella sp. detected in spleen samples from black rats (R. rattus) was closely related to both B. tribocorum and B. elizabethae. The species detected in the Cairo spiny mouse (A. cahirinus) spleen sample was closely related to the zoonotic pathogen, B. elizabethae. These results indicate that Bartonella species are highly prevalent in suburban rodent populations and their ectoparasites in Israel. Further investigation of the prevalence and zoonotic potential of the Bartonella species detected in the black rats and the Cairo spiny mouse is warranted.     

Real‐time PCR detection of Erwinia amylovora on blossoms correlates with subsequent fire blight incidence

Fire blight is the most devastating bacterial disease of rosaceous plants. Forecasting fire blight infections is important to allow for countermeasures that reduce economic damage in pome fruit production. Current computerized forecasting models are solely based on physical factors such as temperature and moisture, but not on the actual presence of the pathogen Erwinia amylovora. Although the inoculum concentration is considered to be crucial for infection and disease outbreak, most current approaches used for identification of fire blight inoculum including morphological, biochemical, serological, and DNA‐based methods are nonquantitative. Based on a real‐time PCR approach previously published, an improved protocol to be used directly on whole bacteria in the field is described. The method allows for early detection and quantification of the pathogen prior to the occurrence of first symptoms. There is a clear correlation between bacterial abundance and subsequent disease development. However, in some cases, no disease symptoms could be observed despite a pathogen load of up to 3·4 × 10⁶ cells per blossom. Integration of the amount of pathogen detected into refined prediction algorithms may allow for the improvement of applied forecasting models, finally permitting a better abatement of fire blight.     

Detection of phloridzin in strawberries (Fragaria x ananassa Duch.) by HPLC-PDA-MS/MS and NMR spectroscopy

The phenolic profile of strawberry fruits (Fragaria x ananassa Duch., Rosaceae) was investigated by high-performance liquid chromatography with photodiode array detection. A peak displaying retention time and UV spectral data identical to those of phloridzin (phloretin 2'-O-beta-d-glucoside), a dihydrochalcone glucoside so far considered characteristic of apples, was monitored. For further characterization, crude extracts of strawberries were purified on polyamide, and the target compound was isolated by preparative and analytical HPLC. Structure elucidation was performed on the basis of APCI- and ESI-MS in the negative ion mode as well as by 1D and 2D NMR spectroscopy using authentic phloridzin for comparison. The d-configuration of the sugar moiety was established by HPLC analysis of the corresponding acyclic 1-deoxy-1-(N-acetyl-alpha-methylbenzylamino)alditol acetate. Apart from its chemotaxonomic relevance, this first report on the occurrence of phloridzin in strawberries is of particular interest for the authenticity control of strawberry products such as juices, jams, and fruit preparations since phloridzin has so far been used for the detection of fraudulent admixtures.