Canine fetal echocardiography: correlations for the analysis of cardiac dimensions

The aim of this study was to develop regression models for correlation of canine fetal heart development with body size to characterize normal development or suggest cardiac anomalies. Twenty clinically healthy pregnant bitches, either brachycephalic and non-brachycephalic, were examined ultrasonographically. Transabdominal fetal echocardiography was conducted every 4 days from the beginning of cardiac chambers differentiation until parturition. Ten cardiac parameters were measured: length, width and diameter of the heart; heart area; left and right ventricular dimensions; left and right atrial dimensions; and aortic and pulmonary artery diameter. Femoral length, biparietal diameter and abdominal cross-sectional area were also recorded. Regression equations were developed for each parameter of fetal body size, and linear and logarithmic models were compared. The model with the highest correlation coefficient was chosen to produce equations to calculate relative dimensions based on the correlations. Only the left-ventricular chamber differed between the two racial groups. Biparietal diameter was the independent parameter that produced the highest correlation coefficient for the most fetal cardiac dimensions, although good correlations were also observed using femoral length and abdominal cross-sectional area. Heart width and heart diameter were used as surrogates of cardiac development, as these measurements showed the best statistical correlation. Quantitative evaluation of fetal cardiac structures can be used to monitor normal and abnormal cardiac development.     

Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology

Tendons regenerate poorly due to a dense extracellular matrix and low cellularity. Cellular therapies aim to improve tendon repair using mesenchymal stem cells and tenocytes; however, a current limitation is the low proliferative potential of tenocytes in cases of severe trauma. The purpose of this study was to develop a method useful in veterinary medicine to improve the differentiation of Peripheral Blood equine mesenchymal stem cells (PB-MSCs) into tenocytes. PB-MSCs were used to study the effects of the addition of some growth factors (GFs) as TGFβ3 (transforming growth factor), EGF2 (Epidermal growth factor), bFGF2 (Fibroblast growth factor) and IGF-1 (insulin-like growth factor) in presence or without Low Level Laser Technology (LLLT) on the mRNA expression levels of genes important in the tenogenic induction as Early Growth Response Protein-1 (EGR1), Tenascin (TNC) and Decorin (DCN). The singular addition of GFs did not show any influence on the mRNA expression of tenogenic genes whereas the specific combinations that arrested cell proliferation in favour of differentiation were the following: bFGF2 + TGFβ3 and bFGF2 + TGFβ3 + LLLT. Indeed, the supplement of bFGF2 and TGFβ3 significantly upregulated the expression of Early Growth Response Protein-1 and Decorin, while the use of LLLT induced a significant increase of Tenascin C levels. In conclusion, the present study might furnish significant suggestions for developing an efficient approach for tenocyte induction since the external administration of bFGF2 and TGFβ3, along with LLLT, influences the differentiation of PB-MSCs towards the tenogenic fate.     

Immunological and pathological investigations in equine experimental uveitis

Background

The pathogenic mechanism of equine recurrent uveitis (ERU) is still poorly defined and many variations between experimental animal models and spontaneous disease exist.

Objectives

The aim of our study was to investigate if Th17 cell-mediated response plays role in the pathogenesis of the used experimental model in horses and to reveal its pathological findings.

Methods

Experimental uveitis was induced in 6 healthy horses. The concentrations of retinal autoantigen CRALBP and IL-17 were measured using ELISA in aqueous humor and vitreous body of the 12 inflamed eyes as well as in 12 control non-inflamed eyes taken from 6 horses in slaughter house. After centrifugation of the two eye media, smears were prepared and cytological investigation was performed. Tissue specimens were taken from all eye globes and were submitted to histopathological investigation.

Results

CRALBP and IL-17 concentrations were significantly elevated in eye media of horses with experimental uveitis in comparison with controls. Cytological and histopathological findings corresponded to the changes characteristic of chronic immune-mediated inflammation with mononuclear cell infiltration of uvea, choroid, retina, and eye media as well as severe retinal destruction.

Conclusions

Our study demonstrated the involvement of the retinal autoantigen CRALBP as well as IL-17 in the pathogenesis of experimental uveitis in horses. These findings suggests that this experimental uveitis in horses may serve as a suitable animal model for investigation of IL-17- mediated immune response during spontaneous autoimmune uveitis in horses as well as in humans.

     

Determination of biogenic amines in aerobically stored beef using high-performance thin-layer chromatography densitometry

The aim of this study was to develop a high-performance thin-layer chromatography densitometry method to simultaneously evaluate levels of different biogenic amines (BAs) in beef stored aerobically at 4°C. Steaks from M. longissimus thoracis et lumborum muscles were overwrapped with polyvinyl chloride film and stored for 0, 4, 7, and 10 days. For this purpose, the solvent system and pH for the derivatization of BAs with dansyl chloride were optimized. Moreover, this method was also validated for linearity, limits of detection and quantification, precision, and recovery. As found using this method in beef samples, spermine content decreased significantly after 7 days of storage, and tyramine and cadaverine were not detected until day 10.     

Pyruvate kinase is necessary for Brucella abortus full virulence in BALB/c mouse

Brucellosis, caused by a facultative intracellular pathogen Brucella, is one of the most prevalent zoonosis worldwide. Host infection relies on several uncanonical virulence factors. A recent research hotpot is the links between carbon metabolism and bacterial virulence. In this study, we found that a carbon metabolism-related pyruvate kinase (Pyk) encoded by pyk gene (locus tag BAB_RS24320) was associated with Brucella virulence. Determination of bacterial growth curves and resistance to environmental stress factors showed that Pyk plays an important role in B. abortus growth, especially under the conditions of nutrition deprivation, and resistance to oxidative stress. Additionally, cell infection assay showed that Pyk is necessary for B. abortus survival and evading fusion with lysosomes within RAW264.7 cells. Moreover, animal experiments exhibited that the Pyk deletion significantly reduced B. abortus virulence in a mouse infection model. Our results elucidated the role of the Pyk in B. abortus virulence and provided information for further investigation of Brucella virulence associated carbon metabolism.     

Content and distribution of different phosphorus forms in broiler chickens

To understand the background value of phosphorus in chickens, the quantitative distribution of different phosphorus forms, including total phosphorus (TP), free phosphate (FP) and phospholipid (PL), in viscera, blood and bones of broiler chickens was investigated. Results showed that phosphorus contents exhibited significant differences in different parts of chickens. TP content of breast and thigh meat was over 5.0 g/kg, in which most of the phosphorus was in the form of water‐soluble phosphates. TP content in viscera was higher than that in meat, and spleen was observed to contain the highest amount of phosphorus (10.0 g/kg). In all tested organs, FP and PL contents in liver were the highest, ranging between 1207–1989 and 81–369 mg/kg respectively. TP content in chicken bone was in the range of 52 716–136 643 mg/kg, and FP content in the bone was relatively lower than that in chicken meat.     

Estimation of the standardized ileal digestible lysine requirement for primiparous pregnant sows

This experiment was conducted to determine the optimal standardized ileal digestible lysine (SID Lys) level in diets fed to primiparous sows during gestation. A total of 150 (Landrace × Large White) crossbred gilts (weighing 149.9 ± 3.1 kg) were fed gestation diets (12.55 MJ of ME/kg) containing SID Lys levels of 0.43, 0.52, 0.60, 0.70 or 0.80% respectively. Gilts were fed 2.0 kg/day from day 1 to 80 and 3.0 kg/day from day 80 to 110 of gestation respectively. Gilts were allocated to treatments based on their body weight on the day of breeding. Weight gain from day 80 to 110 increased with increasing dietary SID Lys levels (p = 0.044). Fitted broken‐line (p = 0.031) and quadratic plot (p = 0.047) analysis of body weight gain indicated that the optimal SID Lys level for primiparous sows was 0.70 and 0.69% respectively. During gestation, neither backfat thickness nor loin eye area was affected by dietary SID Lys level. Increasing dietary Lys had no effect on the litter size at birth or pigs born alive per litter. Litter weight at birth was not affected by dietary SID Lys level. The litter weight variation at birth quadratically decreased with increasing dietary SID Lys (p = 0.021) and was minimized at 0.70% dietary SID Lys. Gilts fed the 0.70% SID Lys diet had the highest dry matter (p = 0.031) and protein (p = 0.044) content in colostrum. On day 110 of gestation, gilts fed the 0.70% SID Lys diet tended to have the highest serum prolactin (p = 0.085) and serum insulin (p = 0.074) levels. The data demonstrate that the optimal dietary SID Lys was 0.70% for pregnant gilts, which is similar to the recommendation of 0.69% that was estimated by the NRC (2012).     

Two single nucleotide polymorphisms in the promoter of the ovine myostatin gene (MSTN) and their effect on growth and carcass muscle traits in New Zealand Romney sheep

Myostatin is a negative regulator of muscle growth and development in mammals, and variation in ovine myostatin gene (MSTN) has been demonstrated to be associated with variation in the muscularity of sheep. Polymerase chain reaction–single‐stranded conformational polymorphism (PCR–SSCP) was used to look for single nucleotide polymorphisms (SNPs) in a 304‐bp amplicon from the promoter region of ovine MSTN. Sequence analyses revealed two previously identified SNPs (c.?2449G/C and c. ?2379T/C) that resulted in three haplotypes (H₁ (c.[?2449G; ?2379C]), H₂ (c.[?2449C; ?2379C]) and H₃ (c.[?2449G; ?2379T]). The effect of these SNPs on growth and carcass traits was investigated in 357 NZ Romney lambs. General linear mixed‐effect models revealed that sheep with the genotype c.?2449GC had a higher loin meat yield (p = 0.032) and proportion loin yield (p = 0.028), than those with the genotype c.?2449GG. The genotype c.?2379CC was associated with an increase in three weight traits: birthweight (p = 0.003), tailing weight (p = 0.009) and weaning weight (p = 0.028), when compared with the genotype c.?2379TC, but it was not found to have an association with growth rate. This suggests that c.?2379T/C has an effect that originates at, or before birth. Haplotype H₃ was associated with a decrease in birthweight (p = 0.002), tailing weight (p = 0.003) and weaning weight (p = 0.011). Haplotype H₂ was associated with increased loin yield (p = 0.012) and proportion loin yield (p = 0.002). The SNPs may have value as genetic markers for improved Romney breeding.