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.     

Analysis of world strains of Anaplasma marginale using major surface protein 1a repeat sequences

Anaplasma marginale is a tick-borne pathogen of cattle that causes the disease bovine anaplasmosis worldwide. Major surface proteins (MSPs) are involved in host-pathogen and tick-pathogen interactions and have been used as markers for the genetic characterization of A. marginale strains and phylogenetic studies. MSP1a is involved in the adhesion and transmission of A. marginale by ticks and varies among geographic strains in the number and sequence of amino-terminal tandem repeats. The aim of this study was to characterize the genetic diversity of A. marginale strains collected from countries in North and South America, Europe, Asia, Africa and Australia, inclusive of all continents. In this study, we characterized 131 strains of A. marginale using 79 MSP1a repeat sequences. These results corroborated the genetic heterogeneity of A. marginale strains in endemic regions worldwide. The phylogenetic analyses of MSP1a repeat sequences did not result in clusters according to the geographic origin of A. marginale strains but provided phylogeographic information. Seventy-eight percent of the MSP1a repeat sequences were present in strains from a single geographic region. Strong (> or =80%) support was found for clusters containing sequences from Italian, Spanish, Chinese, Argentinean and South American strains. The phylogenetic analyses of MSP1a repeat sequences suggested tick-pathogen co-evolution and provided evidence of multiple introductions of A. marginale strains from various geographic locations worldwide. These results contribute to the understanding of the genetic diversity and evolution of A. marginale and tick-pathogen interactions.     

Phosphorylation of inositol 1,4,5-triphosphate receptor 1 during in vitro maturation of porcine oocytes

During fertilization in mammalian species, a sperm-induced intracellular Ca²⁺ signal ([Ca²⁺]_i) mediates both exit of meiosis and oocyte activation. Recently, we demonstrated in mouse oocytes that the phosphorylation levels of inositol 1,4,5 trisphosphate receptor type1 (IP₃R1), the channel responsible for Ca²⁺ release and oscillations during fertilization, changed during maturation and fertilization. Therefore, we examined the expression and phosphorylation of IP₃R1 during in vitro maturation of pig oocytes. Here, our present study shows that expression of IP₃R1 protein did not change during maturation, although the phosphorylation status of the receptor, specifically at an MPM-2 epitope, did. We found that while at the beginning of maturation IP₃R1 lacked MPM-2 immunoreactivity, it became MPM-2 reactive by 24 h and reached maximal reactivity by 36 h. Interestingly, the acquisition of MPM-2 reactivity coincided with the activation of p34^cdc2 kinase and mitogen-activated protein kinase (MAPK), which are involved in meiotic progression. Following completion of maturation, inactivation of MAPK by U0126 did not affect IP₃R1 phosphorylation, although inactivation of p34^cdc2 kinase by roscovitine dramatically reduced IP₃R1 phosphorylation. Neither inhibitor affected total expression of IP₃R1. Altogether, our results show that IP₃R1 undergoes dynamic phosphorylation during maturation and this might underlie the generation of oscillations at fertilization.     

Differentially expressed genes identified through RNA‐seq with extreme values of principal components for beef fatty acid in Nelore cattle

The aim of this study was to identify differentially expressed genes (DEG) in the Longissimus thoracis muscle of Nelore cattle related to fatty acid (FA) profile through RNA sequencing and principal component analysis (PCA). Two groups of 10 animals each were selected containing PC1 and PC2 extreme DEG values (HIGH × LOW) for each FA group. The intramuscular fat (IMF) was compared between cluster groups by ANOVA, and only the sum of monounsaturated FA (MUFA) and ω3 showed significant differences (p < .05). Interestingly, the highest percentage (95%) of phenotypic variation explained by the sum of the first two PC was observed for ω3, which also displayed the lowest number of DEG (n = 1). The lowest percentage (59%) was observed for MUFA, which also revealed the largest number of DEG (n = 66). Since only MUFA and ω3 exhibited significant differences between cluster groups, we can conclude that the differences observed for the remaining groups are not due to the percentage of IMF. Several genes that have been previously associated with meat quality and FA traits were identified as DEG in this study. The functional analysis revealed one KEGG pathway and eight GO terms as significant (p < .05), in which we highlighted the purine metabolism, glycolytic process, adenosine triphosphate binding and bone development. These results strongly contribute to the knowledge of the biological mechanisms involved in meat FA profile of Nelore cattle.     

Effects of exogenous amylase on the in vitro digestion kinetics of whole-crop maize silages made from flint or dent grain type at different phenological stages grown in tropical condition

The effect of exogenous amylase on the in vitro rumen digestion kinetics of whole-crop maize silage made from dent (RB9004) or flint grain type (RB9308) was evaluated at different phenological stages: soft dough (SOD), early dent (EAD), ½ milkline (½M) and ¾ milkline (¾M). Forage was harvested from 70 to 110 days after sowing. Two rumen-cannulated cows receiving or not exogenous amylase (0.7 g/kg dry matter—DM, provided to achieve 396 kilo Novo units of amylase activity/kg of TMR DM) were used as donor of ruminal fluid. The in vitro gas production kinetics was evaluated according to a dual-pool logistic model. The chemical composition and gas production kinetics were affected by the hybrid and phenological stages. The flint hybrid had lower range for chemical analysis among physiological stages. Harvesting at ½M and ¾M improved DM content, bromatological composition and silage quality parameters compared to dent or flint types. Amylase (i) increased methane (CH₄) production and in vitro dry matter digestibility (IVDMD) in ½M stage, (ii) improved digestion kinetics by reducing lag time and increasing total gas production and fermentation rates of non-fibrous carbohydrates (NFC) and fibrous carbohydrates (FC), and (iii) increased extent and fermentation rate of NFC and increased fermentation rate of FC fraction in whole-crop maize silages produced from dent or flint types in all phenological stages. Harvesting between ½M and ¾M is the best phenological stage to improve chemical composition and silage quality parameters. Exogenous amylase showed improvements on fibre digestion of silages at ½M and ¾M phenological stages in both grain types of corn.     

Concomitant turkey herpesvirus-infectious bursal disease vector vaccine and oil-adjuvanted inactivated Newcastle disease vaccine administration: consequences for vaccine intake and protection

Hatchery vaccination protocols in day-old chicks are designed to provide early priming and protection against several poultry diseases including, but not limited to, Marek's disease (MD), infectious bursal disease (IBD), and Newcastle disease (ND). The constraint of concomitant administration of live MD and IBD vaccines plus ND inactivated oil-adjuvanted vaccines (IOAVs) requires improvements in vaccine technology. Single-needle concomitant subcutaneous (SC) application of IBD/MDV and killed NDV vaccine and the use of viral vectors for expression of immunogenic proteins are a current trend in the industry. The objective of this work was to assess the compatibility of a turkey herpesvirus (HVT)-infectious bursal disease (vHVT-IBD) vector vaccine applied simultaneously with IOAV and to evaluate the consequences for vaccine intake, the need for additional immunizations with the respective vaccines, and protection. Five separate trials were performed using double- and/or single-needle injectors. The levels and persistence of vaccine intake, serologic response, vHVT-IBD virus combination with the MD Rispens strain, and/or live NDV vaccination were also assessed. Histopathology and PCR at injection sites showed adequate vaccine intake detected up to 44 days postvaccination. Serologic evidence of vaccine priming was observed, and all vaccinated groups differed (P < 0.05) from the control at different time points. MD, NDV, and IBD protection results after concomitant double-shot single-needle vaccination were near 85%, 95%, and 100%, respectively. Taken together the results indicate no deleterious effects on the efficacy of the vHVT-IBD vaccine monitored by vaccine intake, serologic and challenge results, and combinations after concomitant live/killed vaccination, suggesting the suitability of its use in hatchery vaccination. All types of injectors used as well as injection techniques, vaccines injected separately or together, gave the same results.     

Oxytetracycline treatment reduces bacterial diversity of intestinal microbiota of Atlantic salmon

The effect of oxytetracycline (OTC) treatment on intestinal bacterial populations in juvenile Atlantic salmon Salmo salar was evaluated. Oxytetracycline was administered by way of medicated feed to fish held in experimental tanks. Restriction fragment length polymorphism and sequencing of 16S rDNA from isolates were used to analyze the intestinal microbiota before, during, and after OTC administration. The microbiota from untreated fish was more diverse, consisting mainly of Pseudomonas, Acinetobacter, Bacillus, Flavobacterium, Psycrobacter, and Brevundimonas spp. In contrast, the microbiota of the OTC-treated group was characterized by lower diversity and consisted only of Aeromonas, clustering with A. sobria and A. salmonicida. Antibiotic-resistant isolates were identified as Aeromonas spp.; sequencing the resistance determinant showed it to be the tetE gene. Overall, OTC treatment changed the composition of the intestinal microbiota of Atlantic salmon, as evidenced by a reduction in bacterial diversity. These results support the current concern that antibiotic treatment can facilitate the proliferation of opportunistic bacteria by eradicating competing microorganisms.     

ASSESSING CIRCLE OF WILLIS BLOOD CIRCULATION IN DOGS WITH TRANSCRANIAL COLOR-CODED DUPLEX SONOGRAPHY

Insonation of Circle of Willis by transcranial Doppler duplex color sonography is described in 30 healthy dogs with 15 weighing <33 lb and 15 weighing >33 lb. Imaging was via a temporal window to explore the rostral, middle, and caudal cerebral arteries on both the left and right-hand sides; and through an suboccipital window to study the basilar artery. Normal mean values of the peak systolic velocity (PSV), end diastolic velocity, mean velocity, resistance index (RI), and pulsatility index (PI) were characterized and compared with those obtained in previous studies. There was significant differences in the PSV, RI, and PI in the rostral cerebral artery between dogs weighing <33 vs. >33 lb. Mean PSV was higher in weighing over 33 lb, whereas the mean resistive index and mean PI were lower in these dogs.     

Obstetric and fetal outcomes in dystocic and eutocic sows to an injection of exogenous oxytocin during farrowing

Sixty hybrid Yorkshire-Landrace penned sows, 30 with eutocic farrowing and 30 experiencing a dystocic parturition, were studied to evaluate the obstetric and neonatal outcomes to low doses of oxytocin administered at advanced stages of parturition. Animals in each group were randomly subdivided into 2 subgroups: 15 eutocic and 15 dystocic sows received oxytocin 0.083 IU/kg (equivalent to 1 IU/12 kg body weight), administered intramuscularly after the delivery of the 5th piglet; the other 15 eutocic and 15 dystocic sows received saline solution intramuscularly at the same time. Oxytocin decreased the number of intrapartum deaths by approximately 50% (P = 0.002). No piglet was born dead from the saline- and oxytocin-treated eutocic sows. The highest viability score was observed among piglets born to eutocic sows treated with oxytocin. In summary, this dose schedule would help to decrease the number of stillbirths in both eutocic and dystocic farrowing sows.