Serum iron concentration in cattle with endotoxaemia.

The objective of this study was to examine whether serum iron (Fe) concentration is useful as a prognostic biomarker for cows with acute coliform mastitis (ACM). Our study was composed of determining the reproducibility of serum Fe concentration as a prognostic criterion in cows with ACM (Study 1) and clarifying the sequential changes in serum Fe concentration in cattle that received endotoxin (Study 2). Seventy-seven cows with (n = 47) or without (n = 30) ACM were enrolled in Study 1. The proposed diagnostic cut-off value of serum Fe concentration indicating a poor prognosis of ACM based on the analysis of the receiver operating characteristic curves was < 31.5 µg/dL. Ten young cattle aged 176.8 ± 23.7 days were enrolled in Study 2. Five young cattle received endotoxin (LPS group) and the remaining five received physiological saline (control group). Blood collections were carried out before endotoxin challenge (pre), and 0.5, 1, 2, 4, 8, 12, 24, and 48 h after the challenge. As a result, a significant decrease in serum Fe concentration was not observed until 24 h after endotoxin challenge. Because in cows with clinical ACM it is difficult to know the time course after infection, the alteration in serum Fe concentrations alone may be an insufficient prognostic criterion.

Serum iron concentration as a marker of inflammation in young cows that underwent dehorning operation.

This study aimed to assess the usefulness of serum iron (Fe) concentration as a marker of inflammation caused by the dehorning operation. Five young Holstein cows aged 205.0 ± 10.7 days and weighing 207.2 ± 24.1 kg underwent the dehorning operation. Blood samples were withdrawn before dehorning (pre) and at time periods of t=0.5, 2, 4, 6, 8, 12, 24, and 48 hr. The serum amyloid A (SAA) concentration was significantly high at t=48 hr (P<0.01). The serum Fe concentration significantly decreased, reaching 90.0 ± 36.4 µg/dl at t=24 hr (P<0.001). Therefore, serum Fe concentration showed significant and negative correlation with SAA concentration (r2=0.500, P<0.01). In conclusion, serum Fe concentration is a useful marker of inflammation in young cows that have undergone the dehorning operation.

Sequential changes in serum zinc concentrations in calves with experimentally induced endotoxin shock measured by the particle-induced X-ray emission method.

The aim of the present study was to measure changes in the serum concentrations of some elements in endotoxin-challenged calves using a particle-induced X-ray emission analysis and to screen for elements useful as diagnostic markers. The results obtained revealed that serum Zn concentrations were more accurate diagnostic markers for detecting endotoxin shock in calves than other elements. Serum Zn level in endotoxin-challenged calve was significantly lower from 8 to 12 hr after the endotoxin challenge than pre-challenge values. In addition, serum Zn concentrations in calves from 4 to 24 hr after endotoxin challenges were significantly lower than those of control. Our results indicate that serum Zn concentration has potential as diagnostic markers for detecting inflammation in calves with endotoxin shock.

Plasma lead, silicon and titanium concentrations are considerably higher in green sea turtle from the suburban coast than in those from the rural coast in Okinawa, Japan.

The purpose of this study was to compare the concentration of trace elements in the plasma of sea turtles that inhabited the suburban (Okinawa Main Island, n=8) and the rural coast (Yaeyama Island, n=57) in Okinawa, Japan. Particle induced X-ray emission allowed detection of 20 trace and major elements. The wild sea turtles in the suburban coast in Okinawa were found to have high concentrations of Pb, Si and Ti in the plasma when compared to the rural area but there were no significant changes in the Al, As and Hg concentrations. These results may help to suggest the status of some elements in a marine environment. Further, monitoring the plasma trace and major element status in sea turtles can be used as a bio-monitoring approach by which specific types of elements found here could indicate effects that are related to human activities.

Plasma endotoxin activity in Eastern grey kangaroos (Macropus giganteus) with lumpy jaw disease.

Progressive pyogranulomatous osteomyelitis involving the mandible or maxilla of captive macropods, referred to as "Lumpy jaw disease (LJD)", is one of the most significant causes of illness and death in captive macropods. The aim of the present study was to evaluate the relationship between the severity of LJD and plasma endotoxin activity in kangaroos. Plasma samples obtained from moderate (n=24) and severe LJD (n=12), and healthy kangaroos (n=46), were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. Plasma endotoxin activity was measured using the Limulus amebocyte lysate (LAL)-kinetic turbidimetric (KT) assay. Plasma endotoxin activity was higher in kangaroos with severe LJD (0.199 ± 0.157 EU/ml) than in those with moderate LJD (0.051 ± 0.012 EU/ml, P<0.001) and healthy controls (0.057 ± 0.028 EU/ml, P<0.001). Our results suggest that the severity of LJD in captive macropods may be related to the plasma endotoxin activity.

Particle-Induced X-ray Emission Analysis of Zierum Trace and Major Elements in Cattle with Acute Coliform Mastitis.

The aim of the present study was to examine the applicability of the direct determination of trace and major element concentrations in serum samples collected from Holstein dairy cattle with acute coliform mastitis (n = 53) compared with a healthy control group (n = 39). Twenty-eight elements (Na, Mg, Al, Si, S, Cl, K, Ca, Ti, V, Cr, Mn, Fe, Ce, Ni, Cu, Zn, Ga, As, Se, Br, Rb, Sr, Y, Zr, Nb, Mo, and Pb) were detected by particle-induced X-ray emission (PIXE). Significant differences were observed in serum K, Fe, Zn, and Br concentrations, but not in those of the remaining twenty-four elements. Furthermore, serum Fe concentrations (0.751 ± 0.583 µg/ml, n = 18) were significantly lower in dairy cattle with a poor prognosis than in those with a good prognosis (0.945 ± 0.393 µg/ml, n = 35, P < 0.05) and healthy controls (1.458 ± 0.391 µg/ml, n = 39, P < 0.01). We proposed a diagnostic cut-off point for serum Fe concentrations of <0.82 µg/ml based on receiver operating characteristic (ROC) curves in order to identify cattle with a poor prognosis. The results of the present study indicated that assessing the elemental composition of serum, particularly iron, is a promising prognostic tool for determining the outcomes of cattle with severe acute coliform mastitis.

Plasma endotoxin activity in kangaroos with oral necrobacillosis (lumpy jaw disease) using an automated handheld testing system.

The aim of the present study was to evaluate the reliability and effectiveness of directly determining endotoxin activity in plasma samples from kangaroos with lumpy jaw disease (LJD, n=15) and healthy controls (n=12). Prior to the present study, the ability of the commercially available automated handheld portable test system (PTS(TM)) to detect endotoxin activity in kangaroo plasma was compared with that of the traditional LAL-kinetic turbidimetric (KT) assay. Plasma samples, which were obtained from endotoxin-challenged cattle, were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. The performance of the PTS(TM) was not significantly different from that of the traditional LAL-based assay. The data obtained using PTS(TM) correlated with those using KT (r(2)=0.963, P<0.001). These findings indicated that the PTS(TM) is applicable as a simplified system to assess endotoxin activity in macropods. In the present study, we demonstrated the diagnostic value of plasma endotoxin activity in kangaroos with systemic inflammation caused by oral necrobacillosis and identified plasma endotoxin activity as a sensitive marker of systemic inflammation in kangaroos with LJD. Based on ROC curves, we proposed a diagnostic cut-off point for endotoxin activity of >0.22 EU/ml for the identification of LJD. Our results indicate that the assessment of plasma endotoxin activity is a promising diagnostic tool for determining the outcome of LJD in captive macropods.

Evaluation of a portable test system for assessing endotoxin activity in raw milk.

The aim of the present study was to compare endotoxin activities detected in raw milk samples obtained from cattle by a commercially available portable test system (PTS) and traditional microplate limulus amebocyte lysate (LAL)-based assay, which determined activities using a kinetic turbidimetric (KT) assay. Raw milk samples were obtained from 53 and 12 dairy cattle without and with clinical mastitis, respectively. Comparison between the KT and PTS was performed by the Friedman test. The Pearson product moment correlation coefficients were calculated to evaluate associations between any two continuous variables. Linear regression model analysis was also performed to obtain the equation describing the relationship between PTS and KT assay. The endotoxin activities detected in 200- or 400-fold diluted milk samples were similar between PTS and KT assay, whereas a significant difference was observed in 100-fold diluted milk (P<0.001). The results obtained from 200- (r(2)=0.778, P<0.001) and 400-fold diluted milk samples (r(2)=0.945, P<0.001) using PTS correlated with those using KT assay. The median milk endotoxin activities in Gram-positive and Gram-negative clinical mastitis cows were 0.655 and 11,523.5 EU/ml, respectively. The results of the present study suggest that PTS as a simple and easy test to assess endotoxin activity in raw milk is efficient, simple and reproducible.

Detecting endotoxin activity in bovine serum using an automated testing system.

The aim of the present study was to compare the ability of the commercially available portable test system (PTS(TM)) to detect endotoxin activity in bovine serum, with that of the traditional LAL-kinetic turbidimetric (KT) and chromogenic (KC) assays. Prior to testing, serum samples, which were obtained from endotoxin-challenged cattle, were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. The performance of the PTS(TM) was not significantly different from that of the traditional LAL-based assays. The results using PTS(TM) correlated with those using KT (r(2)=0.963, P<0.001) or KC assays (r(2)=0.982, P<0.001). Based on these findings, the PTS(TM) could be applied as a simplified system to assess endotoxin activity in bovine serum.

Profiles of serum amino acids to screen for catabolic and inflammation status in calves with Mycoplasma bronchopneumonia.

The aim of the present study was to investigate the relationships between serum amino acid profiles in normal and calves with Mycoplasma bronchopneumonia. Serum free amino acid concentrations in serum obtained from 34 calves with or without Mycoplasma bronchopneumonia were determined by high-performance liquid chromatography. The calves with Mycoplasma were characterized by significantly lower total amino acid and total essential amino acid concentrations and molar ratios of branched-chain amino acid (BCAA) to aromatic amino acid (BCAA/AAA) and BCAA to tyrosine (BTR), and by a significantly higher molar ratio of serine phosphorylation (SPR). The proposed diagnostic cutoffs for BCAA/AAA, BTR and SPR in serum based on ROC analysis for detection of catabolic states associated with Mycoplasma bronchopneumonia were set at <1.75, <2.86 and >0.85, respectively. Our results suggest that determining the profiles of amino acids, especially BTR and SPR, could provide useful diagnostic information in terms of predicting protein catabolism in Mycoplasma bronchopneumonia.