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.     

Effect of Spinacia oleraceae L. and Perilla frutescens L. on Antioxidants and Lipid Peroxidation in an Intervention Study in Healthy Individuals

Daily consumption of fruits and vegetables is frequently recommended to prevent several diseases. This health-promoting effect is considered to be in part due to the antioxidant content of fruits and vegetables and their ability to decrease oxidative stress. To investigate whether the ingestion of preparations from spinach or perilla, two carotenoid-rich leafy vegetables, is followed by an increase in carotenoid concentration and/or affects parameters of oxidative stress in human blood plasma. 12 healthy volunteers ingested a perilla- or spinach preparation for 10 days (5 mg lutein/d). We quantified antioxidant levels in plasma, kinetics of lipid peroxidation, MDA concentration, and total antioxidative capacity of plasma. We observed a significant increase in lutein content and a moderate increase (n.s.) in β-carotene content in human blood plasma after consumption of spinach or perilla. The markers of lipid peroxidation tended to decrease, but no influence on antioxidative capacity of plasma could be detected. The high lutein content of perilla caused a more pronounced increase of lutein compared to spinach. Both vegetables seem to be able to influence lipid peroxidation in a beneficial manner.     

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.     

Alternative inoculum sources for fire blight: the potential role of fruit mummies and non‐host plants

Fire blight is the most damaging bacterial disease in apple production worldwide. Cankers and symptomless infected shoots are known as sites for the overwintering of Erwinia amylovora, subsequently providing primary inoculum for infection in the spring. In the present work, further potential sources of inoculum were investigated. Real‐time PCR assays covering a 3‐year‐period classified 19·9% of samples taken from fruit mummies as positive. Bacterial abundance in fruit mummies during autumn, winter and spring was up to 10⁹ cells per gram of tissue and correlated well with later infection rates of blossoms. Blossoms of non‐host plants growing close to infected trees were also shown to be colonized by E. amylovora and to enable epiphytic survival and propagation of bacteria. The results indicate a potential role of fruit mummies and buds in overwintering and as a source of primary inoculum for dissemination of the pathogen early in the growing season. Non‐host blossoms may also serve as an inoculum source in the build‐up of the pathogen population. Both aspects may contribute significantly to the epidemiology of E. amylovora. The significance of infected rootstocks as an inoculum source is also discussed. Fruit mummies might be used to determine pathogen pressure in an orchard before the beginning of the blooming period.     

Lignin als biologische Barriere gegen Schimmelpilze in Innenräumen

Many different fungi grow indoors as mold in the presence of moisture. Some of these fungi can cause allergic or toxic reactions, while a few may cause infections in susceptible individuals. A comprehensive treatment of this complex topic would take volumes. Therefore many groups search for alternative management strategies of these fungi. It has been tested if the growth of nine fungi (Alternaria alternata, Botrytis cinerea, Cladosporium herbarum, Fusarium culmorum, F. solani, F. verticillioides, Mucor sp., Penicillium sp. and Trichoderma sp.) which are common in interior buildings could be inhibited by mixing lignin into wall plaster. In maltextract agar lignin concentrations above 10 % inhibited mould growth except for Penicillium sp. However, none of the mould species showed and degradation of lignin. Further investigations on a lignin-plaster-agar, which contained only plaster as nutrient and had a high pH at 8, showed no inhibition on the tested fungi (Alternaria alternata, Fusarium verticillioides, Mucor sp., Trichoderma sp. and Penicillium sp.) at concentrations up to 5 %. Higher lignin concentrations lead to insufficient quality of building materials, e. g. cracking. Furthermore the color of the building materials turned from greyish-white in an exotherm reaction into brown, become not any more suitable for construction purposed.     

Microbial response of soils with organic and conventional management history to the cultivation of Bacillus thuringiensis (Bt)-maize under climate chamber conditions

An experiment was carried out in a climate chamber to analyse if Bt-maize may cause particular changes in soils with different levels of microbial biomass and activity due to long-term management history. Among the soils selected, the ones managed organically for 30 years exhibited twice the microbial biomass and 2.6 times the dehydrogenase activity (DHA) of the soil from a field with long-term conventional maize monoculture. Soils were cultivated twice in a row with Bt-maize, its near-isogenic line and a conventional breeding line. We tested the hypotheses that (a) soil microbial biomass and activity are affected by the cultivation of Bt-maize and that (b) the influence of Bt-maize depends on the level of soil microbial biomass and activity. Shoot and root yield and shoot C-content of Bt-maize were higher than the ones of the near-isogenic line. DHA under Bt-maize was 6 % higher, and the metabolic quotient for CO₂ (qCO₂) was 9 % lower than under its near-isogenic line, giving some support to hypothesis (a). No significant interactions of the soils and the varieties used were found in this study, thus hypothesis (b) was not confirmed, and soils with different microbial biomass and activity appear to react in a similar way to the cultivation of Bt-maize.     

Moderne Entmottung

Ohne Zusammenfassung     

Investigation into association and expression of PLCz and COX-2 as candidate genes for boar sperm quality and fertility

Phospholipase C zeta (PLCz) and cyclooxygenase isoenzyme type 2 (COX-2) are important in spermatogenesis, but their effect has not yet confirmed in pigs. Therefore, this study was aimed to analyse their association with sperm quality and fertility and to identify the mRNA and protein expression in boars reproductive tissues. DNA samples from 231 Pietrain (PI) and 109 Pietrain × Hampshire (PIHA) pigs with records of sperm quality [sperm concentration (SCON), motility, semen volume, plasma droplet and abnormal spermatozoa rate] and fertility (non-return rate and number of piglet born alive) traits were available. A SNP in non-coding region of PLCz g.158 A > C was associated with SCON (p < 0.05) in PIHA population while the polymorphism of COX-2 g.68 G > A in 3' UTR was not associated with any traits. For mRNA and protein expression study, a total of six boars were divided into two groups with G-I and G-II, where G-I was characterized for relatively better sperm quality. Both genes expressed higher in reproductive tissues compared with non-reproductive tissues. Phospholipase C zeta mRNA expressed higher in testis (p < 0.01), all parts of epididymis and spermatozoa from G-I, while COX-2 expressed higher in testis (p < 0.05), head and body of epididymis (p < 0.01), and spermatozoa from G-II boar. Both proteins were localized in Leydig cells and spermatozoa. These results might shed light on roles of these genes in spermatogenesis as candidate for boar sperm quality and fertility, but still the lack of association across populations should be considered.