Putative regulation mechanism for the MSTN gene by a CpG island generated by the SINE marker Ins227bp.

BACKGROUND: A single nucleotide polymorphism (SNP) in the first intron of the myostatin gene (MSTN) is associated with aptness of elite Thoroughbreds to race over sprint, middle or long distances. This intronic marker (g.66493737 T ≻ C), a short interspersed nuclear element (SINE) of 227 bp (Ins227bp) insertion polymorphism in the MSTN promoter, and the adjacent SNP BIEC2-417495 have not been studied for their association with racing aptness of the average Thoroughbreds raced in countries with lower status of the racing industry. This study investigated these markers regarding their prevalence and association with performance in common race horses. Markers were genotyped by amplification refractory mutation system-quantitative PCR (ARMS-qPCR) or amplicon melting. Furthermore, we asked whether the Ins227bp marker might theoretically regulate the expression of myostatin by generating a novel target for DNA methylation or by changing binding sites for transcription factors. Putative sites for DNA methylation or binding of transcription factors were predicted by MethPrimer and by the softwares JASPAR, MatInspector and UniPROBE, respectively. RESULTS: Pairwise linkage disequilibrium between g.66493737 T ≻ C and Ins227bp was high (r (2) = 0.93). A lower linkage was determined for g.66493737 T ≻ C and BIEC2-417495 (r (2) = 0.69) as well as for BIEC2-417495 and Ins227bp (r (2) = 0.76). The estimated frequencies for the presence of Ins227bp (I) indel and the C alleles at g.66493737 T ≻ C and BIEC2-417495 were 0.46, 0.47 and 0.43, respectively. Heterozygotes represented the most abundant genotype at each locus. The best racing distance (BRD) was significantly different between the homozygotes of each SNP (p = 0.01 to 0.03). C allele homozygotes at BIEC2-417495 or g.66493737 T ≻ C, as well as Ins227bp homozygotes earned most money on a mean distance ranging from 1211 to 1230 m. Heterozygotes earned most money on races over 1690 to 1709 m. The BRD for the T/T carriers at both SNP loci and for the SINE-free genotype was 1812 to 1854 m. Other performance parameters were not significantly different between the genotypes, except of the relative success score (RSS). The RSS was significantly slightly better on a distance of ≤ 1300 m for all carriers of the C allele and the Ins227bp compared to homozygous T genotypes and SINE-negative horses (p = 0.037 to 0.046). For distances of more than 1300 m the RSS was not significantly different between genotypes. In silico assessment indicated that the Ins227bp promoter insertion might have generated a CpG island and a few novel putative binding sites for transcription factors. CONCLUSIONS: All three target polymorphisms (Ins227bp, g.66493737 T ≻ C, BIEC2-417495) are suitable markers to assess the ability of non-elite Thoroughbreds to race at short or longer distances. The CpG island generated by Ins227bp may cause training-induced silencing of MSTN expression.

Monitoring of inflammatory blood biomarkers in foals with Rhodococcus Equi pneumonia during antimicrobial treatment.

Rhodococcus equi (R. equi), a gram-positive facultative intracellular pathogen, is a common cause of pneumonia in foals and represents a major cause of disease and death. The aim of the present study was to investigate the time-depended changes in White Blood Cells (WBC), basophils (Baso), neutrophils (Neu), lymphocytes (Lymf), monocytes (Mon), eosinophils (Eos), platelet (PLT) counts, fibrinogen (Fbg) concentration, interferon (IFN-α, IFN-γ) and interleukins (IL-2 and IL-10) in foals with clinical R. equi pneumonia. The main treatment was with azithromycin-rifampicin for 14 days. Blood was sampled prior to, 7 and 14 days after starting therapy. Treatment was associated with significantly decreased counts of WBC, (25.6 ± 6.7 and 14.2 ± 2,7 × 103/ml), Neu (18.6 ±6.2 and 10.7 ± 3.1 × 103/ml), Mon (1.5 ± 0.5 and 0.9 ± 0.2 × 103/ml) and Fbg (539 ± 124 and 287 ± 26 g/dl) between day 0 and day 14. IL-2 and IL-10 concentrations were significantly increased (P = 0.028, P = 0.013, respectively) after treatment, whereas IFN-α and IFN-γ concentrations were not. The diagnostic potentials of INF-α, INF-γ, IL-2 and IL-10 per se seems not very high, however, the study suggests that the activity change of selected interleukins in the course of the disease may be associated with amelioration. We concluded that patterns of serum concentration changes of INF-α, INF-γ, IL-2 and IL-10 may help in the study of the innate immune response in foals during infection and treatment of R. equi pneumonia.

Chloride and lactate as prognostic indicators of calf diarrhea from eighty-nine cases.

IMPORTANCE: Deaths due to neonatal calf diarrhea are still one of the most critical problems of cattle breeding worldwide. Determining the parameters that can predict diarrhea-related deaths in calves is especially important in terms of prognosis and treatment strategies for the disease. OBJECTIVE: The primary purpose of this study was to determine mortality rates and durations, survival status, and predictive prognosis parameters based on vital signs, hematology, and blood gas analyses in neonatal diarrheic calves. METHODS: The hospital automation system retrospectively obtained data from 89 neonatal diarrheic calves. RESULTS: It was found that 42.7% (38/89) of the calves brought with the complaint of diarrhea died during hospitalization or after discharge. Short-term and long-term fatalities were a median of 9.25 hours and a median of 51.50 hours, respectively. When the data obtained from this study is evaluated, body temperature (°C), pH, base excess (mmol/L), and sodium bicarbonate (mmol/L) parameters were found to be lower, and hemoglobin (g/dL), hematocrit (%), lactate (mmol/L), chloride (mmol/L), sodium (mmol/L) and anion gap (mmol/L) parameters were found to be higher in dead calves compared to survivors. Accordingly, hypothermia, metabolic acidosis, and dehydration findings were seen as clinical conditions that should be considered. Logistic regression analysis showed that lactate (odds ratio, 1.429) and CI- (odds ratio, 1.232) concentration were significant risk factors associated with death in calves with diarrhea. CONCLUSIONS AND RELEVANCE: According to the findings obtained from this study, the determination of lactate and Cl- levels can be used as an adjunctive supplementary test in distinguishing calves with diarrhea with a good prognosis.

Daily variability of forced oscillometry parameters in horses suffering recurrent airway obstruction, a pilot study.

The aim of the study was to analyse the day-to-day variability of the respiratory resistance (R(rs)) and the reactance (X(rs)) in 5 horses in a status of remission of recurrent airway obstruction by forced oscillometry system (FOS). Furthermore, the effects of stabling, outdoor and room temperature and humidity on these parameters were determined. Mean R(rs) at oscillation frequencies 1, 1.5, 2 and 3 Hz were all significantly higher in the morning than in the afternoon, while X(rs) was not significantly affected by time of the day. R(rs) was significantly different on various sampling days at all frequencies, in the morning as well as in the afternoon. X(rs) showed the same significant differences for frequencies greater than 1 Hz. Overall, R(rs) showed a smaller variability than X(rs). Neither outdoor temperature, nor room temperature affected R(rs). This also applied to X(rs) except for the measurements at 2 Hz that were significantly (p = 0.044) affected by outdoor temperature. Turning out these horses at paddock for day and night caused a significant lower mean R(rs) that when kept at stable for all night. While R(rs) showed a very weak positive frequency dependency, X(rs) clearly showed a positive frequency dependence for all horses. Resonance frequency (f(res)) was between 2 and 4 Hz. These studies showed that FOS generated parameters are affected by environmental and management factors and thus only after well-designed standardized procedures FOS could be a useful diagnostic tool for the diagnosis and monitoring of equine respiratory disorders.

Changes in serum mineral concentrations, biochemical and hematological parameters in horses with pica.

The aim of this study was to compare hematological, some biochemical parameters, and serum trace element concentrations in horses with or without pica. Fifteen horses with pica (group I) and another 15 healthy horses without pica (group II) were used. The hematological parameters were not changed between the two groups. In group I, hemoglobin values were lower than those of group II. However, the difference in hemoglobin values between the two groups was not significant (P > 0.05). Serum iron and copper concentrations and the copper/zinc ratio were lower in group I than those of group II (P < 0.05). The other biochemical parameters were not found to be statistically different between the two groups (P > 0.05). It was concluded that serum iron and copper deficiency may play an important role for the etiology of pica in horses. Prophylactic use of iron and copper supplements in horses may be beneficial to prevent pica.