Markers In Cattle Research Articles

Discrepancies of bovine haptoglobin concentrations between serum and plasma using two different anticoagulants and a colorimetric assay based on peroxidase activity.

Haptoglobin (Hp) is an emerging diagnostic marker in cattle, and knowledge of suitable sample types and measurement methods is important. The aims of this study were to compare the results of a colorimetric assay (CA) and an ELISA for bovine Hp using serum, EDTA plasma, and lithium-heparinized (LH) plasma, respectively, and to assess the diagnostic potential for puerperal metritis. In experiment 1, Hp was measured in pooled aliquots of serum (n = 10), EDTA plasma (n = 10), and LH plasma (n = 10) of 100 healthy fresh lactating dairy cows from 10 farms using both the CA and the ELISA. In experiment 2, five healthy and five cows with acute puerperal metritis were sampled, and Hp was determined using both assays for all three sample types. In experiment 3, aliquots of serum and LH plasma from cows in different lactation stages were transferred into plain, EDTA-coated, and LH-coated tubes and mixed before colorimetric analyses. Distilled water was also placed into each tube type and treated similarly. Plasma samples measured with the CA showed on average 2.3 (EDTA) and 2.5 (LH) times higher Hp concentrations compared with serum, whereas no differences were seen with the ELISA results between sample types. Based on a clinical cut-off value, both methods differentiated sick from healthy cows. Haptoglobin measurements with the ELISA were less precise compared with CA measurements due to high dilutions. No influence of the anticoagulants on the CA was observed. Due to measurement discrepancies between serum and plasma, CAs for bovine Hp based on peroxidase activity should be performed with serum, or specific reference ranges for plasma samples should be established. In this study, CA results obtained with LH plasma were more precise than results obtained with EDTA plasma. Both the CA and the ELISA are suitable diagnostic tools for the diagnosis of puerperal metritis, but CA measurements were more precise in this study.

A comprehensive analysis of the effects of DGAT1 K232A polymorphism on milk production and fertility traits in Holstein Friesian and Jersey cows reared in Türkiye

Abstract. Research on the diacylglycerol acyltransferase 1 (DGAT1) K232A marker in cattle shows inconsistent results across regions, largely due to small sample sizes, limited genetic variation, and data restricted to few lactations, which complicates establishing a reliable genotype–phenotype correlation. This research aimed to determine the effect of the K232A polymorphism of the bovine DGAT1 gene on milk production and quality traits in dairy cattle. We used 1104 cattle, including 828 Holstein Friesian and 276 Jersey cows. The analysis utilized extensive data from six lactations of cows raised on four commercial dairy farms. We genotyped the population using the polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) technique and Sanger sequencing for verification. We then evaluated the 305 d and test-day milk yields as well as fat and protein yields and percentages. The number of inseminations per conception and calving ease were also assessed as reproduction indices. Genotype–phenotype associations were quantified using linear mixed models. The AA genotype was absent in Jersey cows, and the heterozygous genotype was predominant in both breeds. The K232A marker was significantly associated with test-day milk yield, fat, and protein content in Jersey cows. Further, it substantially affected the fat percentage of milk in Holstein Friesian cows (p<0.001). We found that the KK genotype is highly desirable for milk quality and especially fat content. This comprehensive assessment demonstrated that the KK genotype of the DGAT1 K232A polymorphism significantly influenced fat and protein contents in dairy cattle.

Identification of novel bovine leukocyte antigen alleles and association of BoLA-DRB3.2*020:02:01 with resistance to Theileria orientalis infection in crossbred Kedah-Kelantan cattle: a pilot study.

The bovine leukocyte antigen (BoLA) gene is a significant genetic part of the immune system and has been used as a disease marker in cattle. In this study, we detected Theileria orientalis, T. sinensis, Anaplasma marginale, Anaplasma platys, Candidatus Mycoplasma haemobos and Trypanosoma evansi by PCR amplification and sequencing of the amplicons. The allelic association of the BoLA-DRB3.2 gene with blood pathogen disease resistance and susceptibility in 87 Kedah-Kelantan x Brahman (KKB) and 38 Bali cattle was determined by Fisher's exact test and Cochran Mantel Haenszel (CMH) correction test. Sequence-based typing of the BoLA-DRB3.2 gene identified 43 alleles (27 previously reported alleles and 16 novel alleles) across the two cattle breeds. Alignment analysis of the 16 novel alleles revealed 90.7-95.8% and 85-92% nucleotide and amino acid identities, with the reference allele, BoLA-DRB3*016:01 cDNA clone NR-1. BoLA-DRB3*009:02 (25.6%) and BoLA-DRB3*036:01 (36%) were the most frequent alleles in KKB and Bali cattle, respectively. In KKB cattle, BoLA-DRB3*020:02:01 was significantly associated with resistance to T. orientalis whereas *007:01 and *009:02 were significantly associated with resistance to C. Mycoplasma haemobos. Also, DRB3*017:01 was associated with susceptibility to T. orientalis in KKB cattle. In the Bali cattle, BoLA-DRB3*015:01 was found to be a genetic marker of susceptibility to C. Mycoplasma haemobos infection. Therefore, this study identified BoLA-DRB3.2 alleles associated with resistance and susceptibility to T. orientalis infection in KKB cattle and susceptibility to C. Mycoplasma haemobos infection in Bali cattle for the first time. Therefore, this study suggests that these BoLA-DRB3 resistance alleles could be used as candidate markers for selection, whereas susceptibility alleles could be used as candidate markers for culling in the beef industry.

Genetic diversity of DGAT1 gene linked to milk production in cattle populations of Ethiopia

BackgroundDiacylglycerol acyl-CoA acyltransferase 1 (DGAT1) has become a promising candidate gene for milk production traits because of its important role as a key enzyme in catalyzing the final step of triglyceride synthesis. Thus use of bovine DGAT1 gene as milk production markers in cattle is well established. However, there is no report on polymorphism of the DGAT1 gene in Ethiopian cattle breeds. The present study is the first comprehensive report on diversity, evolution, neutrality evaluation and genetic differentiation of DGAT1 gene in Ethiopian cattle population. The aim of this study was to characterize the genetic variability of exon 8 region of DGAT1 gene in Ethiopian cattle breeds.ResultsAnalysis of the level of genetic variability at the population and sequence levels with genetic distance in the breeds considered revealed that studied breeds had 11, 0.615 and 0.010 haplotypes, haplotype diversity and nucleotide diversity respectively. Boran-Holstein showed low minor allele frequency and heterozygosity, while Horro showed low nucleotide and haplotype diversities. The studied cattle DGAT1 genes were under purifying selection. The neutrality test statistics in most populations were negative and statistically non-significant (p > 0.10) and consistent with a populations in genetic equilibrium or in expansion. Analysis for heterozygosity, polymorphic information content and inbreeding coefficient revealed sufficient genetic variation in DGAT1 gene. The pairwise FST values indicated significant differentiation among all the breeds (FST = 0.13; p ≤ 0.05), besides the rooting from the evolutionary or domestication history of the cattle inferred from the phylogenetic tree based on the neighbourhood joining method. There was four separated cluster among the studied cattle breeds, and they shared a common node from the constructed tree.ConclusionThe cattle populations studied were polymorphic for DGAT1 locus. The DGAT1 gene locus is extremely crucial and may provide baseline information for in-depth understanding, exploitation of milk gene variation and could be used as a marker in selection programmes to enhance the production potential and to accelerate the rate of genetic gain in Ethiopian cattle populations exposed to different agro ecology condition.

Development and evaluation of a real-time PCR panel for the detection of 20 immune markers in cattle and sheep

The establishment of a panel of immune markers is of paramount importance to understand the different transcription patterns of infectious diseases in livestock. The array of commercially available immunological assays for cattle and sheep is currently limited, due to the lack of antibodies for these species. Even though SYBR Green based real time quantitative PCR (qPCR) is the most commonly used method to study cytokine transcription in ruminants, a lack of standardization impairs its implementation in the study of different ruminant diseases. In order to obtain reliable qPCR results, several variables need to be considered: choice of reference genes for optimal normalization, variation of annealing temperature among primer sets, and assay specificity and sensitivity.In this study, we developed and validated a panel of immune markers in bovine and ovine samples using SYBR Green based qPCR in a cost-effective way with multiple primer sets optimised to amplify at a common thermal cycling temperature. Twenty primer sets were designed to quantify immune markers (IL-1b, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-15, IL-18, IL-23, TNF-α, IFN-γ, IFN-α, Ki-67, NFkB-65, TLR-3, TLR-4, TLR-8 and Rig-1) in ovine and bovine templates. For optimal normalization and selection of suitable reference genes, primer sets that measure the transcription of five reference genes were also included in the panel. The amplification efficiency, linearity and specificity was validated for all target genes. Optimal amplification conditions were achieved in both ovine and bovine samples for all gene targets, with the exception of Ki67. Relative quantification studies were performed on ovine and bovine mRNA obtained from sheep peripheral blood mononuclear cells (PBMCs) stimulated with three different treatments (PMA/Ionomycin, Concanavalin A (Con A) and pokeweed mitogen (PWM)). Pokeweed and ConA efficiently induced gene transcription of most of the targeted genes, while PMA/Ionomycin showed a weaker induction. Finally, we further assessed usability of our panel by running it on bovine monocyte derived dendritic cells (MoDCs) stimulated with different vaccines. Results confirmed the induction of a specific pro-inflammatory gene transcription pattern by rabies vaccine, which resembles the one occurring during viral infection.Altogether, we validated the efficiency and usability of an extended real-time PCR panel that gives the possibility to rapidly measure a broad spectrum of ovine and bovine immune markers by using a single set of reagents and protocol thus representing a valid and cost-effective tool for research purposes.

Ultrastructural and molecular characteristics of Setaria species based on sequence analysis of genomic and mitochondrial gene markers in cattle (Bos taurus) and buffaloes (Bubalus bubalis) from Iran

The aim of the present study was to investigate the ultrastructural characteristics and genetic diversity of Setaria parasites from cattle (n=696) and buffalo (n= 522) from Khuzestan province of Iran and to compare them with available data from other countries/regions by sequences analysis of the 12S Rdna and the mitochondrial cytochrome C oxidase subunit I (cox1) genes. Based on SEM (Scanning Electron Micrographs) and light microscopy, all the isolated worms were identified as Setaria labiatopapillosa. Our results showed that 12.3% of cattle were infected with Setaria spp., while no infection was found in buffaloes. The maximal prevalence was observed in cattle younger than one year old. The prevalence rate was not influenced by the season of the year or gender. Comparison of the obtained sequences from Setaria with sequences of Setaria spp. from GenBank confirmed that all samples belong to the species S. labiatopapillosa. The phylogenetic tree constructed using cox1 and 12S rDNA genes of several other filarial nematodes showed that the Khuzestan isolates share a common branch with S. labiatopapillosa from other regions. Intra-specific variation was observed in 12S rDNA but not in cox1. In conclusion, our results indicating that S. labiatopapillosa is the main species involved in the spread of setarial infection in south-west of Iran and the identified worms corresponded mostly to worms that reported previously throughout other continents.

Detection of exosome markers in cattle infected with bovine leukaemia virus (BLV)

Detection of exosome markers in cattle infected with bovine leukaemia virus (BLV)

Systemic inflammatory and stress markers in cattle and sheep submitted to different reproductive procedures

ABSTRACT: Previous studies have evaluated the effects of different reproductive procedures on discomfort markers in sheep and cattle. Such studies may help stimulate the adoption of techniques that are more beneficial for animal welfare. However, markers that are commonly used to evaluate discomfort are highly influenced by external factors. To overcome this, several systemic markers can be evaluated to more precisely identify stress, pain, and inflammation. Such markers include cortisol, acute phase proteins, bradykinin, and substance P. We aimed to review the potential markers of stress, pain, and inflammation, and discuss how and when they are regulated after different stimuli related to reproductive procedures in cattle and sheep. Furthermore, we aimed to review how reproductive procedures with different degrees of invasiveness cause stress and provide information that may help develop strategies to limit animal discomfort.

Animal science meets agricultural practice: Preliminary results of an innovative technical approach for exhaled breath analysis in cattle under field conditions

In human and veterinary medicine, the search for exhaled volatiles as potential markers for disease detection has gained increasing attention because exhaled breath analysis bears the advantages of being continuous, non-invasive and reveals instantly available results. The aim of this study was to introduce a technical set-up for breath analysis in bovines. The technical set-up allows (i) CO2-controlled sampling and preconcentration of alveolar gas and (ii) direct breath gas analysis via proton transfer reaction time-of-flight mass spectrometry (PTR-ToF-MS) in parallel. While alveolar gas sampling enables the analysis by gas chromatography-mass spectrometry for unequivocal substance identification, PTR-ToF-MS ensures real-time identification of confounding signals within one measurement. Applicability of the system to conscious cattle and reliability of the signals obtained under field conditions were tested at eight dairy farms. This first testing phase in veterinary practice confirmed that the set-up was well tolerated by each of the cows and that reliable measured values (i.e., raw data) were gained at different respiratory rates. Inspiration could be clearly distinguished from expiration, thus providing the basis for excluding ambient air contamination. Consecutive measurements under standardised conditions in two bovines aged 5–6 months revealed effects of time of day, food intake and handling that need to be evaluated in-depth in future studies with larger groups of animals. As result of these proof-of-concept trials, the introduced technical set-up can be recommended for future studies evaluating exhaled markers in cattle. Breath gas analysis offers a new research area in veterinary medicine and is one step forward towards refining large animal studies, even under field conditions.

Hematological, biochemical and oxidative stress studies of lumpy skin disease virus infection in cattle

ABSTRACTLumpy skin disease (LSD) is an important disease of cattle. This study was conducted to determine alterations in hematological, biochemical and oxidative stress markers in cattle that have been naturally infected with LSD virus (LSDV). Blood samples and skin nodular lesions were collected from clinically infected, recovered and healthy animals. Quantitative real-time PCR was used to screen for Capripoxvirus DNA in samples from clinically infected animals. Hematological, biochemical and oxidative stress markers were measured. LSDV nuclic acids were detected in the collected samples using PCR. Hematological results revealed erythrocytosis, thrombocytopenia and leukopenia in infected cattle. Biochemical analyses showed that total protein and globulin levels were significantly elevated, while albumin and glucose were significantly reduced in these cattle. Aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), creatine phosphokinase (CPK), and creatinine levels were markedly elevated. Moreover, serum levels of reduced glutathione (GSH) were markedly lowered, whereas lipid peroxidation (MDA) and inflammatory cytokines (IL-4 and TNF-α) were elevated. Recovered cattle exhibited significant amelioration of the alterations resulting from LSDV infection. The results of this study suggest that LSDV infection induces changes in hematological and biochemical parameters and stimulates oxidative stress; these findings may be helpful for choosing a good strategy for rapidly detecting and diagnosing LSDV infection.

Global model fitting to compare survival curves for faecal indicator bacteria and ruminant-associated genetic markers.

To compare decay profiles of ruminant- and cattle-associated molecular markers for faecal contamination and Escherichia coli, facilitating their correct application in water quality studies. We generated decay profiles for cultivable E.coli, a general Bacteroidales genetic marker (GenBac3), ruminant markers (CF128, Rum2Bac) and cattle markers (CowM2, CowM3) using faeces-seeded mesocosms, and selected best fitting models for each decay profile. Global model fitting tested for differences between decay profiles. After normalizing for initial concentration, decay curves differed significantly between E.coli and all genetic markers except CowM3. Decay curves for CF128 differed from GenBac3 and Rum2Bac, but Rum2Bac and GenBac3 decay profiles did not differ. Despite similar survival profiles for some markers, highly varied initial concentrations affected time to nondetection. Decay curves and time until nondetection differed among markers from the same host. However, the Rum2Bac and GenBac3 markers had similar decay profiles and could potentially be investigated further for source allocation using the ratio method. As the use of genetic markers for microbial source tracking becomes increasingly common, caution is necessary. Both the shape of decay curves and time to nondetect may differ depending on the marker selected, resulting in possible misinterpretation of results and precluding application of a 'ratio method' of source allocation.

Identification of lactoferrin and glutamate receptor-interacting protein 1 in bovine cervical mucus: A putative marker for oestrous detection.

Accurate detection of oestrus is important for artificial insemination. The aim of this study was to identify oestrous-specific bovine cervical mucus proteins that could be used to determine the optimal time for artificial insemination. Non-oestrous and controlled internal drug release (CIDR)-induced oestrous-stage mucus proteins were purified and subjected to surface-enhanced laser desorption/ionization time-of-flight mass spectrometry, sodium dodecyl sulphate polyacrylamide gel electrophoresis and MALDI-TOF/TOF. Among differentially expressed proteins, lactoferrin (LF) and glutamate receptor-interacting protein 1 (GRIP1) showed a twofold increase during the CIDR-induced oestrous stage compared to the levels in non-oestrous stage in bovine cervical mucus. The RT-PCR, Western blotting and immunohistochemistry results showed that LF and GRIP1 expression was significantly increased during the oestrous stage in the uterus. This study demonstrated that bovine LF and GRIP1 exist during the oestrous stage, but not during the non-oestrous stage, suggesting that cervical mucus LF and GRIP1 are useful oestrous detection markers in cattle.

NTPDase and 5'-nucleotidase as inflammatory markers in cattle naturally infected by Eurytrema coelomaticum.

The aim of this study was to evaluate seric NTPDase and 5'nucleotidase activities of cattle naturally infected by Eurytrema coelomanticum, as well as to correlate them to histopathological lesions in the pancreas and the degree of parasitism. Blood samples and pancreas of 51 bovines were collected on a slaughterhouse in Southern Brazil: 33 from cattle naturally infected by E. coelomanticum (the Group A), and 18 from uninfected animals (the Group B). Infected animals showed an average of 532 parasites per pancreas. In the pancreatic histology, ducts displayed hyperplasia, stenosis, proliferation of fibrous tissue, and interstitial inflammatory infiltration of lymphocytes. The serum from infected animals showed an increase in NTPDase activity when ATP was used as substrate (P<0.001). For the ADP substrate, there was no difference between groups regarding NTPDase activity (P=0.37), as well as 5'-nucleotidase activity (P=0.27). Correlating NTPDase activity (ATP substrate) with the degree of histopathological lesions (rho=0.66, P<0.001) and the parasitic load on the pancreas (rho=0.65, P<0.001), a positive correlation was observed. Similar results were found between the degree of histopathological lesions and NTPDase activity (ADP substrate; rho=0.29, P=0.03), and 5'nucleotidase activity (rho=0.35, P=0.01). Based on the results of NTPDase and 5'nucleotidase enzymes in cattle naturally infected by E. coleomanticum, it is possible to suggest that these enzymes are involved in the modulation of inflammation, and they can act as markers of inflammatory response.

Accuracy of prediction of simulated polygenic phenotypes and their underlying quantitative trait loci genotypes using real or imputed whole-genome markers in cattle.

BackgroundMore accurate genomic predictions are expected when the effects of QTL (quantitative trait loci) are predicted from markers in close physical proximity to the QTL. The objective of this study was to quantify to what extent whole-genome methods using 50 K or imputed 770 K SNPs (single nucleotide polymorphisms) could predict single or multiple QTL genotypes based on SNPs in close proximity to those QTL.MethodsPhenotypes with a heritability of 1 were simulated for 2677 Hereford animals genotyped with the BovineSNP50 BeadChip. Genotypes for the high-density 770 K SNP panel were imputed using Beagle software. Various Bayesian regression methods were used to predict single QTL or a trait influenced by 42 such QTL. We quantified to what extent these predictions were based on SNPs in close proximity to the QTL by comparing whole-genome predictions to local predictions based on estimates of the effects of variable numbers of SNPs i.e. ±1, ±2, ±5, ±10, ±50 or ±100 that flanked the QTL.ResultsPrediction accuracies based on local SNPs using whole-genome training for single QTL with the 50 K SNP panel and BayesC0 ranged from 0.49 (±1 SNP) to 0.75 (±100 SNPs). The minimum number of local SNPs for an accurate prediction is ±10 SNPs. Prediction accuracies that were based on local SNPs only were higher than those based on whole-genome SNPs for both 50 K and 770 K SNP panels. For the 770 K SNP panel, prediction accuracies were higher than 0.70 and varied little i.e. between 0.73 (±1 SNP) and 0.77 (±5 SNPs). For the summed 42 QTL, prediction accuracies were generally higher than for single QTL regardless of the number of local SNPs. For QTL with low minor allele frequency (MAF) compared to QTL with high MAF, prediction accuracies increased as the number of SNPs around the QTL increased.ConclusionsThese results suggest that with both 50 K and imputed 770 K SNP genotypes the level of linkage disequilibrium is sufficient to predict single and multiple QTL. However, prediction accuracies are eroded through spuriously estimated effects of SNPs that are distant from the QTL. Prediction accuracies were higher with the 770 K than with the 50 K SNP panel.

Identification of functional consequence of a novel selection signature in CYP11b1 gene for milk fat content in Bubalus bubalis

Genomic selection for traits of economic importance is an emerging approach carrying tremendous potentials. Many of polygenic traits as milk fat, protein and yield have been characterize at genomic level and important selection signatures have been identified. Cytochrome P450 enzymes are potential loci for affecting many of dairy capabilities. Present study was conducted for genomic dissection of CYP11b1 gene in riverine buffaloes and seven genetic variations were identified. Out of these, one novel polymorphism (p.A313T) was found well associated with milk fat %age. AB genotyped buffaloes were found to have higher milk fat %age (8.9%) for this loci. p.A313T was further validated at larger data set by restriction digestion using CviAII enzyme. Functional consequences of this locus were also predicted by studying three dimensional structure of CYP11b1 protein. For this purpose, 3D protein model was predicted by homology modeling, secondary structural attributes were determined, signal peptide was predicted and a transmembrane helix was also identified. One of polymorphism (p.Y205L) was found in the vicinity of functionally significant F-G loop region, which is the part of protein gets attached to the inner mitochondrial membrane. But this variation could not be associated and needs further investigation. p.A30V, a popular selection marker in cattle, was found in buffaloes as well but could not be associated and might need further confirmation on larger data set. Results of this study illustrate the impending potential of this gene in determining dairy capabilities of buffaloes and might have a role in selection of superior dairy buffaloes.

Variation in clinical markers in cattle naturally infected with bovine tropical theileriosis.

124 cattle naturally infected with Theileria annulata were inspected for clinical markers. Clinical manifestations of general weakness, reduced appetite, pyrexia reaching 40-42°C and lachrymal discharge were seen in more than 75% of the infected animals. Presence of ticks on body, general dullness and depression, recumbency/prostration, oedema of dependent body parts, diarrhea, hypersalivation, pale mucous membranes and yellow colored urine were revealed by more than 50% of the infected animals. Clinical manifestations of dehydration, abdominal distension/ascites, jaundice and haemoglobinuria were revealed by more than 25% of the infected animals less than 25% of the infected animals showed clinical manifestations of exophthalmia, constipation, melena, buccal cavity erosions, congested mucous membranes, nasal discharge and tachypnoea.

Sodium fluorescein as an internal tracer on the location of bovine urine patches in pastures

Sodium fluorescein (SF) was evaluated and validated as an internal marker in cattle for the location of urine patches in pastures. Three trials were carried out aiming at the following: evaluating the effect of dilution and volume of application on SF fluorescence on the environment; testing dosages and duration of excretion of SF administered orally to cattle; and validating the methodology for SF use on the location of cattle urine patches in pastures. The marker was tested in beef cattle kept on pastures under three grazing systems: degraded pasture under extensive management (De), intensively managed dryland pasture with high stocking rate (Id) and intensively managed irrigated pasture with high stocking rate (Ii). Besides the localization of urination sites, the number of urinations/animal/d and the area covered by urine were also determined. The residue of SF remained fluorescent in the pasture foliage up to 34 h after urination events, allowing the location of urine patches by a black-light flashlight. There was no difference between grazing systems in the number of urine patches/animal/d during the rainy and dry seasons. The average number of urine patches was approximately 11/animal/d. As expected, the estimated volume and area covered by urine varied according to the stocking density. The chosen SF dose (50 mg kg−1 LW) did not adversely affect the animals when administered once daily during 2 d. However, the same SF dose administered during four consecutive days caused urinary disorder in the animal. The distribution of urine patches was spatially dependent on specific characteristics of the paddock.

Significance of homocysteine and cardiac markers in cattle with hypocalcemia

This study aimed to determine the levels of homocysteine and cardiac markers in cows with hypocalcemia. This research was conducted with a total of 20 cows, including 10 with hypocalcemia and 10 healthy controls. Hypocalcemia was diagnosed by determining low blood calcium (Ca) levels, and a standard hypocalcemia treatment was initiated in those cows with the disease. Blood samples were taken before the treatment and again 12 h after the treatment, and there was no significant difference in the homocysteine levels between the groups. Ca, ionized calcium (Ica), magnesium (Mg), and phosphorus (P) levels were lower, while cardiac troponin I (cTnI), parathormone (PTH), creatine kinase (CK), and creatine kinase-MB (CK-MB) levels were higher before treatment as compared with the control group. Increases in the Ca, Mg, Ica, and P levels, and a decrease in the PTH levels were detected after treatment when compared with those levels before treatment. The CK, CK-MB, and Mg levels also increased, while the Ica level decreased in the after-treatment group as compared with the control group. Therefore, the cTnI, CK, and CK-MB in cows with hypocalcemia were determined to be important diagnostically, but insignificant for prognosis. Homocysteine was not important for either diagnosis or prognosis.

Immunohistochemical Detection of Dendritic Cell Markers in Cattle

Dendritic cells (DCs) are "professional" antigen-presenting cells with a critical role in the regulation of innate and adaptive immune responses and thus have been considered of great interest in the study of a variety of infectious diseases. The objective of this investigation was to characterize the in vivo distribution of DCs in bovine tissues by using potential DC markers to establish a basis for the study of DCs in diseased tissues. Markers evaluated included MHCII, CD208, CD1b, CD205, CNA.42, and S100 protein, the latter 2 being expressed by follicular dendritic cells whose origin and role are different from the rest of hematopoietic DCs. Paraffin wax-embedded tissues from 6 healthy Friesian calves were subjected to the avidin-biotin-peroxidase method, and the most appropriate fixatives, dilutions, and antigen retrieval pretreatments were studied for each of the primary antibodies. The most significant results included the localization of CD208-positive cells not only in the T zone of lymphoid organs but also within lymphoid follicles; CD1b-positive cells were mainly found in thymus and interfollicular areas of some lymph nodes; cells stained with anti-CD205 antibody were scarce, and their location was mainly in nonlymphoid tissues; and CNA.42- and S100 protein-positive cells localized in primary lymphoid follicles and light zones of germinal centers, although showing differences in the staining pattern. Furthermore, MHCII was established as one of the most sensitive markers for any DC of hematopoietic origin. These results increase our understanding of DC immunolabeling and will help in future DC studies of both healthy and diseased tissues.

Oxidant/antioxidant status in cattle with liver cystic echinococcosis

The objective of the present study was to evaluate the changes of antioxidants and oxidative stress markers in cattle with cystic echinococcosis (CE). Thirty cattle with liver CE along with 30 healthy cattle were used for the study. Parasitized cattle presented a significantly higher lipid peroxidation assessed by the malondialdehyde (MDA) compared with healthy animals (P<0.05). A significantly lower erythrocyte superoxide dismutase (SOD) and glucose 6-phosphate dehydrogenase (G6PD), and a significantly higher erythrocyte glutathione peroxidase (GPx) in the parasitized group, were observed when compared with healthy group (P<0.05). No significant differences were observed for serum total antioxidant status (TAS), zinc, copper and iron between parasitized and healthy groups. The results obtained in this study suggest that CE in cattle induces changes in the activity of antioxidant enzymes. These changes render host cells susceptible to oxidants and exaggerate the generation of free radicals with a consequent lipid peroxidation enhancement.