scholarly journals Occurrence of Zearalenone and Its Metabolites in the Blood of High-Yielding Dairy Cows at Selected Collection Sites in Various Disease States

Toxins ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 446
Author(s):  
Wojciech Barański ◽  
Magdalena Gajęcka ◽  
Łukasz Zielonka ◽  
Magdalena Mróz ◽  
Ewa Onyszek ◽  
...  
Keyword(s):  
Dairy Cows ◽  
Limit Of Detection ◽  
Correct Diagnosis ◽  
Sampling Site ◽  
Parent Compound ◽  
External Jugular Vein ◽  
Portal System ◽  
Plant Materials ◽  
Blood Samples ◽  
Hepatic Portal

Zearalenone (ZEN) and its metabolites, alpha-zearalenol (α-ZEL) and beta-zearalenol (β-ZEL), are ubiquitous in plant materials used as feed components in dairy cattle diets. The aim of this study was to confirm the occurrence of ZEN and its selected metabolites in blood samples collected from different sites in the hepatic portal system (posthepatic–external jugular vein EJV; prehepatic–abdominal subcutaneous vein ASV and median caudal vein MCV) of dairy cows diagnosed with mastitis, ovarian cysts and pyometra. The presence of mycotoxins in the blood plasma was determined with the use of combined separation methods involving immunoaffinity columns, a liquid chromatography system and a mass spectrometry system. The parent compound was detected in all samples collected from diseased cows, whereas α-ZEL and β-ZEL were not identified in any samples, or their concentrations were below the limit of detection (LOD). Zearalenone levels were highest in cows with pyometra, where the percentage share of average ZEN concentrations reached 44%. Blood sampling sites were arranged in the following ascending order based on ZEN concentrations: EJV (10.53 pg/mL, 44.07% of the samples collected from this site), ASV (14.20 pg/mL, 49.59% of the samples) and MCV (26.67 pg/mL, 67.35% of the samples). The results of the study indicate that blood samples for toxicological analyses should be collected from the MCV (prehepatic vessel) of clinically healthy cows and/or cows with subclinical ZEN mycotoxicosis. This sampling site increases the probability of correct diagnosis of subclinical ZEN mycotoxicosis.

scholarly journals Can Milk Flow Traits Act as Biomarkers of Lameness in Dairy Cows?

Agriculture ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 227
Author(s):  
Vida Juozaitienė ◽  
Ramūnas Antanaitis ◽  
Gediminas Urbonavičius ◽  
Mingaudas Urbutis ◽  
Saulius Tušas ◽  
...  
Keyword(s):  
Dairy Cows ◽  
San Francisco ◽  
Standard Procedure ◽  
Cortisol Concentration ◽  
Initial Time ◽  
Plateau Phase ◽  
Flow Data ◽  
Flow Meters ◽  
Blood Samples ◽  
Milk Flow

We hypothesized that lameness has an impact on milk flow traits. The aim of the current study was therefore to investigate the relation between lameness and milk flow traits in dairy cows. For this study 73 healthy and 55 cows with lameness were selected. Lameness was diagnosed by a local specialized veterinarian, according to the standard procedure. The blood samples were collected during clinical examination. The milking properties of cows were evaluated twice in a row—during evening and morning milking. The selected cows in the current lactation did not receive veterinary treatment, and correct hoof trimming was not performed at least four weeks before the experiment. The measurements were taken by two electronic mobile milk flow meters (Lactocorder®®, WMB AG, Balgache, Switzerland). Milk flow data were processed using LactoPro 5.2.0 software (Biomelktechnik Swiss). Cortisol concentration was measured with the automated analyzer TOSOH®® AIA-360 (South San Francisco, CA, USA). We found out that milk flow traits can act as biomarkers of lameness in dairy cows. We determined that the milk yield in the first minute of healthy dairy cows was 1.77 kg higher than that of lame cows. The electrical conductivity during the initial time of milking of healthy cows was 0.24 mS/cm lower than that of the lame group. The milking duration of LA cows was 1.07 min shorter and the time of incline in milk flow from 0.5 kg/min till the reach of the plateau phase was longer. The risk of lameness was most clearly indicated by an increase in blood cortisol concentration; if its blood level in cows exceeds 1 µg/dL, the risk of identifying lameness increases 4.9 times.

Comparison of Droplet Digital Polymerase Chain Reaction (ddPCR) and Real-Time Quantitative Polymerase Chain Reaction (qPCR) in Detecting Neonatal Invasive Fungal Infections

2021 ◽  
Vol 11 (3) ◽  
pp. 373-379
Author(s):  
Huitao Li ◽  
Xueyu Chen ◽  
Xiaomei Qiu ◽  
Weimin Huang ◽  
Chuanzhong Yang
Keyword(s):  
Polymerase Chain Reaction ◽  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Fungal Isolation ◽  
Blood Samples ◽  
Fungal Strains ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Invasive fungal infection (IFI) is the leading cause of death in neonatal patients, yet the diagnosis of IFI remains a major challenge. At present, most IFI laboratory diagnostic methods are based on classical, but limited, methods such as fungal isolation and culture and histopathological examination. Recently, quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) technology have been adopted to quantify nucleic-acid identification. In this study, we established qPCR and ddPCR assays for IFI diagnosis and quantification. qPCR and ddPCR were carried out using identical primers and probe for the amplification of 18S rRNA. Assay results for three fungal strains were positive, whereas ten non-fungal strains had negative results, indicating 100% specificity for both ddPCR and qPCR methods. Genomic DNA of Candida albicans was tested after a serial dilution to compare the sensitivity of the two PCR methods. The limit of detection of ddPCR was 3.2 copies/L, which was a ten-fold increase compared with that of the qPCR method (32 copies/L). Blood samples from 127 patients with high-risk factors and clinical symptoms for IFI were collected from a NICU in Shenzhen, China, and analyzed using qPCR and ddPCR. Thirty-four blood samples from neonates had a proven or probable diagnosis of IFI, and 25 of these were positive by qPCR, whereas 30 were positive by ddPCR. Among the 93 blood samples from neonates who had a possible IFI or no IFI, 24 were positive using qPCR, and 7 were positive using ddPCR. In conclusion, ddPCR is a rapid and accurate pan-fungal detection method and provides a promising prospect for IFI clinical screening.

scholarly journals Diagnostic performance of capillary and venous blood samples in the detection of Loa loa and Mansonella perstans microfilaraemia using light microscopy

2021 ◽  
Vol 15 (8) ◽  
pp. e0009623
Author(s):  
Johannes Mischlinger ◽  
Rella Zoleko Manego ◽  
Ghyslain Mombo-Ngoma ◽  
Dorothea Ekoka Mbassi ◽  
Nina Hackbarth ◽  
...  
Keyword(s):  
Linear Models ◽  
Limit Of Detection ◽  
Venous Blood ◽  
Microscopic Analysis ◽  
Sub Saharan Africa ◽  
Loa Loa ◽  
Paired Data ◽  
Female Ratio ◽  
Blood Samples ◽  
Mansonella Perstans

Background Loa loa and Mansonella perstans–the causative agents of loiasis and mansonellosis—are vector-borne filarial parasites co-endemic in sub-Saharan Africa. Diagnosis of both infections is usually established by microscopic analysis of blood samples. It was recently established that the odds for detecting Plasmodium spp. is higher in capillary (CAP) blood than in venous (VEN) blood. In analogy to this finding this analysis evaluates potential differences in microfilaraemia of L. loa and M. perstans in samples of CAP and VEN blood. Methods Recruitment took place between 2015 and 2019 at the CERMEL in Lambaréné, Gabon and its surrounding villages. Persons of all ages presenting to diagnostic services of the research center around noon were invited to participate in the study. A thick smear of each 10 microliters of CAP and VEN blood was prepared and analysed by a minimum of two independent microscopists. Differences of log2-transformed CAP and VEN microfilaraemia were computed and expressed as percentages. Furthermore, odds ratios for paired data were computed to quantify the odds to detect microfilariae in CAP blood versus in VEN blood. Results A total of 713 participants were recruited among whom 52% were below 30 years of age, 27% between 30–59 years of age and 21% above 60 years of age. Male-female ratio was 0.84. Among 152 participants with microscopically-confirmed L. loa infection median (IQR) microfilaraemia was 3,650 (275–11,100) per milliliter blood in CAP blood and 2,775 (200–8,875) in VEN blood (p<0.0001), while among 102 participants with M. perstans this was 100 (0–200) and 100 (0–200), respectively (p = 0.44). Differences in linear models amount up to an average of +34.5% (95% CI: +11.0 to +63.0) higher L. loa microfilaria quantity in CAP blood versus VEN blood and for M. perstans it was on average higher by +24.8% (95% CI: +0.0 to +60.5). Concordantly, the odds for detection of microfilaraemia in CAP samples versus VEN samples was 1.24 (95% CI: 0.65–2.34) and 1.65 (95% CI: 1.0–2.68) for infections with L. loa and M. perstans, respectively. Conclusion This analysis indicates that average levels of microfilaraemia of L. loa are higher in CAP blood samples than in VEN blood samples. This might have implications for treatment algorithms of onchocerciasis and loiasis, in which exact quantification of L. loa microfilaraemia is of importance. Furthermore, the odds for detection of M. perstans microfilariae was higher in CAP than in VEN blood which may pre-dispose CAP blood for detection of M. perstans infection in large epidemiological studies when sampling of large blood quantities is not feasible. No solid evidence for a higher odds of L. loa microfilariae detection in CAP blood was revealed, which might be explained by generally high levels of L. loa microfilaraemia in CAP and VEN blood above the limit of detection of 100 microfilariae/ml. Yet, it cannot be excluded that the study was underpowered to detect a moderate difference.

scholarly journals Development and Clinical Validation of a Droplet Digital PCR Method for Detection of Acinetobacter baumannii and Klebsiella pneumonia in Patients with Suspected Bloodstream Infections

2021 ◽  
Author(s):  
Yang Zheng ◽  
Jun Jin ◽  
Ziqiang Shao ◽  
Jingquan Liu ◽  
Run Zhang ◽  
...  
Keyword(s):  
Blood Culture ◽  
Acinetobacter Baumannii ◽  
Limit Of Detection ◽  
Early Stage ◽  
Bloodstream Infections ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Klebsiella Pneumonia ◽  
Clinical Validation ◽  
Blood Samples

The relatively long turnaround time and low sensitivity of traditional blood culture may delay the effective antibiotic therapy in patients with bloodstream infection (BSI). To reduce the morbidity and mortality of BSI, a rapid and sensitive pathogen detection method is urgently required. Acinetobacter baumannii and Klebsiella pneumonia are two major microorganisms responsible for BSI. Here we reported a novel droplet digital PCR (ddPCR) method that can detect A. baumannii and K. pneumonia in whole blood samples within 4 h, with a specificity of 100% for each strain and limit of detection at 0.93 copies/microliter for A. baumannii and 0.27 copies/microliter for K. pneumonia. Clinical validation in 170 patients with suspected BSIs showed that, compared with blood culture that reported 4 (2.4%) A. baumannii cases and 7 (4.1%) K. pneumonia cases, ddPCR detected 23 (13.5%) A. baumannii cases, 26 (15.3%) K. pneumonia cases, and 4 (2.4%) dual infection cases, including the 11 positive patients reported by blood culture. In addition, the positive patients reported by ddPCR alone (n = 42) had significantly lower serum concentrations of procalcitonin and lactate, SOFA and APACHE II scores, and 28-day mortality than those reported by both blood culture and ddPCR (n = 11), suggesting that patients with less severe manifestations can potentially benefit from the guidance of ddPCR results. In conclusion, our study suggests that ddPCR represents a sensitive and rapid method to identify causal pathogens in blood samples and to guide the treatment decisions in the early stage of BSI.

scholarly journals The effects of different anticoagulants on biochemical parameters in blood of dairy cows

2015 ◽  
Vol 69 (1-2) ◽  
pp. 13-20
Author(s):  
Branislava Belic ◽  
Marko Cincovic ◽  
Maja Dosenovic ◽  
Dragica Stojanovic ◽  
Zorana Kovacevic
Keyword(s):  
Alkaline Phosphatase ◽  
Dairy Cows ◽  
Alkaline Phosphatase Activity ◽  
Total Protein ◽  
Biochemical Parameters ◽  
Biochemical Analysis ◽  
Blood Parameters ◽  
Albumin Level ◽  
Blood Samples ◽  
Holstein Friesian

Routine analysis of blood parameters requires high precision. Therefore a significant number of methods and recommendations has been developed to ensure the obtaining of precise results. The aim of this study was to investigate whether the type of anticoagulant affects the values of biochemical parameters in the blood of dairy cows in relation to the values obtained from the serum. The study was carried out on 20 healthy cows of Holstein- Friesian breed in the second month of lactation. The blood was taken by venipuncture from v.coccigea. There were used five types of vacutainers as follows: for the serum separation, with heparin, EDTA, citrate, and fluoride. Samples from each cow were taken in all five types of vacutainers respectively. There was determined the concentration of: albumin, total protein, glucose, Ca, P, BHB, NEFA, urea, cholesterol, triglycerides, ALT, AST, AP, GGT and bilirubin. The results of the analysis of samples that contained anticoagulants were compared statistically with analysis in serum that was considered to be the control one. It was also calculated the percentage of deviation in average values of concentration of metabolites obtained from samples with different anticoagulants in regard to the serum value. The results of the investigation show that anticoagulants influence the values of biochemical parameters in the blood of cows. In the samples where heparin was used as anticoagulant there was found higher albumin level (deviation 4.1%) as well as total protein (1.4%), but lower value of alkaline phosphatase activity relative to the serum (-33%). In the samples in which EDTA was used there were significantly lower levels of total protein (-5.8%), Ca (-49.6%), P (-17.7%), AP (-32%) and higher value AST (10.6%) compared to serum. In the samples where citrate or fluoride were used as anticoagulants there were found lower values of total protein, albumin, glucose (only citrate), Ca, P, BHB, NEFA, urea (only citrate), cholesterol, AP and GGT (only fluoride) and lower bilirubin when using citrate or higher when using fluoride, in comparison to the serum. Deviations of biochemical parameters measured from blood samples that were in citrate or fluoride were as follows: total protein (-5.3%, -5.2%), albumin (-7%, -5.7%), glucose (only citrate -6.2%), Ca (-55%, -82%), P (-29%, -24%), BHB (-53%, -80%), NEFA (-62.1%, -79 , 4%), urea (only citrate, -25.5%), cholesterol (-28.6%, -28.4%), AP (-38%, -32%), and GGT (only -17 fluoride, 8%) and higher bilirubin (-22.8%, 64.4%). For routine clinical biochemical analysis of blood of cows it is the most reliable to use the samples with heparin as an anticoagulant, because the use of other anticoagulants gives large variations in the values of biochemical parameters in relation to serum.

Pre-calving feeding of rumen-protected rice bran to multiparous dairy cows improves recovery of calcaemia after calving

2016 ◽  
Vol 83 (3) ◽  
pp. 281-288 ◽  
Author(s):  
Javier Martín-Tereso ◽  
Holger Martens ◽  
Carolin Deiner ◽  
Harmen van Laar ◽  
Leo A. den Hartog ◽  
...  
Keyword(s):  
Dairy Cows ◽  
Phytic Acid ◽  
Rice Bran ◽  
Control Diet ◽  
Blood Samples ◽  
Group 3

Dairy cows can have different degrees of hypocalcaemia around calving. Lowering dietary Ca availability before calving can prevent it. Rice bran, treated for lower rumen degradability of phytic acid can reduce dietary availability of Ca. During 3 periods of 3 weeks, 113 multiparous cows calved in a single close-up group, which was fed first a control diet, then 140 g/kg DM of rumen-protected rice bran, and at last the control diet again. Cows joined the group 3 weeks before expected calving date and left it at calving. Blood samples were taken weekly before parturition and 0, 6 and 12 h after calving, as well as 3 and 28 d in lactation. Serum was analysed for Ca, Mg, and P. Rice bran introduction produced a transient serum Ca decrease. Rice bran feeding reduced serum P and its withdrawal reduced serum Mg. Serum Ca at calving, nadir of serum Ca and serum Ca the first 3 d after calving was higher in cows calving during rice bran feeding. Serum P decreased less and recovered faster after calving when cows had been fed rice bran. Rumen-protected rice bran reduced dietary availability of Ca and induced adaptation of Ca metabolism resulting in improved Ca and P homoeostasis at calving.

Adhesion of Malassezia pachydermatis of different growth type to canine epithelial cells

2014 ◽  
Vol 17 (2) ◽  
pp. 365-366 ◽  
Author(s):  
M. Florek ◽  
J. Król ◽  
Z. Staroniewicz ◽  
B. Bażanów
Keyword(s):  
Vitamin E ◽  
Epithelial Cells ◽  
Glutathione Peroxidase ◽  
Dairy Cows ◽  
Clinical Mastitis ◽  
Control Group ◽  
Growth Type ◽  
Malassezia Pachydermatis ◽  
Blood Samples ◽  
Experimental Group

Abstract This study focuses on the effect of parenteral administration of Selenium (Se) and vitamin E on concentration of Se in plasma and the activity of glutathione peroxidase (GPx) in the blood of dairy cows during peripartal period and their effect on the reduction of clinical mastitis. From a 220 individuals Holstein herd in a two-four lactation-gestation cycle the control group (C), 1st (D1) and 2nd (D2) experimental group were selected. Every group consisted of 15 cows in the last phase of the pregnancy. All cows were fed with the diet containing 0.1 mg of Se per kg/DM. The blood samples from vena jugularis were collected approximately 21 days before calving (control sampling), 3 days, 12 days and 21 days after calving. On the day of control sampling and 12 days before calving in D1 group, cows were injected subcutaneously in the sprescapular region with preparation Selevit inj. a.u.v. at the doses of 48.4 mg/head of Se, and 550 IU/head of α-Tocoferol (α-Toc). In D2 group, cows were injected by the same preparation only on 21th day before calving with the same doses of Se and α-Toc. The increase in the concentration of Se in the plasma and activity GPx in blood in D1 group on the 3rd day and 12th day after calving were determined. Increase in plasmatic concentrations α-Toc on 3rd day after calving and reduction of occurrence of clinical mastitis (13.3%) as compared with control group were found

scholarly journals Serum Levels of Neuropeptides in Cows with Left Abomasal Displacement

2018 ◽  
Vol 5 (4) ◽  
pp. 103
Author(s):  
Marlene Sickinger ◽  
Joachim Roth ◽  
Klaus Failing ◽  
Axel Wehrend
Keyword(s):  
Substance P ◽  
Dairy Cows ◽  
Vasoactive Intestinal Polypeptide ◽  
Serum Levels ◽  
Common Disease ◽  
Healthy Controls ◽  
Serum Concentrations ◽  
Blood Samples ◽  
Breed Difference ◽  
Abomasal Displacement

Abomasal displacement (AD) to the left is a common disease in high-yielding dairy cows after parturition. In view of the previously reported changes in tissue neuropeptide concentrations in cows with AD, the primary aim of this study was to evaluate the effect of AD and breed on serum neuropeptide concentrations. For this purpose, blood samples of 33 German Holstein (GH) cows with AD, 36 healthy controls (GH), and 32 healthy German Fleckvieh (GF) cows were collected, and concentrations of substance P (SP), vasoactive intestinal polypeptide (VIP), and interleukin1β (IL-1β) were measured via commercially available ELISA kits. To examine the effect of AD, we compared GH cows with and without AD and observed no significant effects of AD on SP, VIP, or Il-1 β concentrations. To evaluate the effect of breed, we compared healthy GH with healthy GF cows and detected markedly higher VIP serum levels in the healthy GF cows (p < 0.01). No significant differences in SP or IL-1β were detected. According to our results, there seems to be no effect of AD on the serum concentrations of SP, VIP, or IL-1 β. In contrast, there seems to be a breed difference concerning serum VIP concentrations.

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