Blood Samples Of Cows Research Articles

Влияние малых доз ионизирующего излучения при облучении in vitro на показатели крови

Abstract. The purpose is the determination of the degree of influence of small absorbed doses of ionizing radiation on biochemical and hematological parameters during irradiation of peripheral blood samples of cattle in vitro. For the work, blood samples were taken from lactating cows kept under the influence of the background absorbed dose in the Krasnoyarsk Territory, followed by in vitro irradiation at doses of 5, 50, 500 mGy. Methods. Peripheral blood was taken from blood vessels from the tail vein into vacuum tubes with a coagulation activator, in vitro irradiation of samples was carried out on an installation equipped with a Cs-137 source, biochemical studies of blood serum were carried out using a PE-5400UF spectrophotometer. Hematological blood parameters were determined according to generally accepted methods. Results. Ionizing radiation in vitro in doses of 5 mGy, 50 mGy and 500 mGy of peripheral blood samples of cattle affects hematological parameters in different directions, which is characterized by wave-like fluctuations in hemoglobin values, a decrease in the content of erythrocytes and a decrease in the erythrocyte sedimentation rate. It was found that a single irradiation of peripheral blood samples of cows at a dose of 5 mGy reduced the concentration of beta-globulins and creatinine and did not affect the content of total protein and protein fractions. When exposed to ionizing radiation at a dose of 50 mGy, the content of total protein, the level of beta-globulins, creatinine decreased in blood samples, and the content of albumins and alpha-, gamma-globulins did not change. When irradiated at a dose of 500 mGy, the relative content of alpha-globulins, the concentration of ALT and AST decreased, the content of creatinine, total protein, albumins and beta-, gamma-globulins did not change. The stability of alkaline phosphatase, albumins and gamma globulins to ionizing effects at doses of 5 mGy, 50 mGy and 500 mGy has been established. Scientific novelty. The linear dependence of the concentration of alpha-globulins, AST and ALT on the absorbed dose was revealed, the numerical values of the approximation coefficients describing the decrease in concentration and biochemical parameters were determined. The inverse direct linear relationship between creatinine stability and absorbed dose has been established. Practical significance is the established digital values of the approximation coefficients of alpha globulins, AST and ALT can be used in the reconstruction of radiation dose values in the range of 5–50 mGy.

Structure of subopopulations of Aireshire, Holstin and Jersey cattle in the Southern Regions of Russia by CSN2 (rs43703011), CSN3 (rs43703016) and microsatellite locuses (BM1818, BM1824, BM2113, ETH10, ETH225, ETH3, INRA023, SPS115, TGLA126, TGLA122, TGLA227, TGLA53)

During studies using molecular biological methods, the following loci were genotyped: BM1818, BM1824, BM2113, ETH10, ETH225, ETH3, INRA023, SPS115, TGLA126, TGLA122, TGLA227, TGLA53, CSN2 (rs43703011) and CSN3 (rs43703 016) 3000 cow blood samples Ayrshire, Holstein and Jersey breeds from the herds of the Krasnodar and Stavropol territories. On average, the frequency of occurrence of homozygous genotypes for 12 microsatellite loci in Jersey cattle was 0.38, in Holstein and Ayrshire cattle – 0.30 and 0.27, respectively; the highest allelic diversity was found in Holstein cattle (9.0 ± 1.7 alleles per locus). It should be noted that there are significant differences in the genetic profile of Ayrshire cattle. For Ayrshire animals, the following alleles turned out to be typical (but rare in Jersey and Holstein cattle): 270 locus BM1818, 137 locus BM2113, 213 locus ETH10, 119 locus ETH3, 142 locus ETH225, 87 locus TGLA227, 166 locus TGLA53. In the studied subpopulation of Jersey cattle, the frequency of occurrence of the A2A2 genotype of the CSN2 locus (rs43703011) (beta casein) was 0.68, and in the Ayrshire and Holstein groups it was only 0.25 and 0.31, respectively. The BB genotype of the CSN3 locus (rs43703016) (casein kappa) was found in the Jersey group of animals with a frequency of 0.76, and in Ayrshires and Holsteins – 0.04 and 0.15, respectively. The genotyped groups of Jersey, Ayrshire and Holstein animals are significant in volume; we assume that the identified patterns are typical for cattle of these breeds in the south of Russia. We believe that the data obtained can be used in the future in practical work on classifying certain groups of livestock as typical Holstein, Ayrshire or Jersey and when creating herds that produce milk of a certain type (for example, type A2 milk).

The effects of a nonsteroidal anti-inflammatory drug on the behavioural and physiological parameters of beef cows and calves assisted at calving

Non-steroidal anti-inflammatory drugs (NSAIDs) may be administered to the cow or the calf as a pain mitigation strategy to combat the negative impacts of injury or trauma associated with a difficult calving. However, the effects of NSAIDs on beef cows’ and calves’ behaviour related to pain and cow-calf bonding after an assisted calving have not been studied. Therefore, the objective of this study was to investigate the effects of meloxicam administered to beef cows and calves after an assisted calving by measuring differences in behaviours associated with pain and cow-calf bonding, and physiological parameters associated with pain. Twenty-three cow-calf pairs assisted at calving were randomly allocated to two medication groups, meloxicam (Metacam®, 20 mg/ml, 0.5 mg/kg body weight, Boehringer Ingelheim, Ingelheim, Germany) or placebo (0.025 ml/kg). Cow blood samples for serum haptoglobin concentration and thermal images of the vulva were taken within 10 minutes after calving and 24±1 hours after calving. Calf blood samples for L‐lactate concentration and thermal images of the head were taken at birth and images repeated at 24±1 hours after birth. Blood samples for serum IgG concentration were taken at 24±1 hours after birth. After at-birth sample collection and treatment administration, the cow-calf pairs were placed in separate box stalls, and behaviours were video recorded for 24±1 hours. A human approach test and a separation test were performed after the 24±1 hour period. Descriptive statistics, multivariable linear and logistic regression models, and non-parametric tests were used to analyze the data. There were no significant differences between meloxicam and placebo groups for serum IgG concentration, haptoglobin concentration, or changes in temperature in the cows’ vulva or calves’ head. Additionally, there were no significant differences between meloxicam and placebo groups for the cows’ behaviours (i.e., time spent lying in sternal recumbency, standing, feeding, self-grooming, grooming the calf, and frequency of tail flicking, tail raising, and pressing against the box stall). Calves treated with meloxicam were more active (P=0.04) and played more (P=0.01) than placebo calves. No other behaviours (i.e., time spent lying in sternal recumbency, attempting to stand, standing, walking, being in a nursing position, investigating, and frequency of approaching the cow) were significantly different between calves treated with meloxicam or placebo. No significant difference (P=0.83) in bonding behaviours was identified for cows and calves treated with meloxicam or a placebo. The anti-inflammatory effects of meloxicam in calves after an assisted calving may improve calf welfare.

Abortion outbreak in pregnant goats and cows with coinfection of ‘Candidatus Mycoplasma haemobos’ and HoBi-like pestivirus

New emerging diseases, including ‘Candidatus Mycoplasma haemobos’ (‘Ca. M. haemobos’) and HoBi-like pestivirus in central China has been a huge challenge for ruminant production. From July to September 2022, an abortion outbreak affected 14 goat farms and 11 cattle farms in this area. To investigate whether the two pathogens are associated with the disease, samples were collected: Group 1 included 55 goat blood samples with foetal tissue samples and ticks on the skin; Group 2 included nine healthy goat blood samples; Group 3 included 36 cow blood samples with foetal tissue samples and ticks on the skin; and Group 4 included seven healthy cow blood samples. Then, these samples were analysed by serology, PCR, sequence analysis, and identification. A total of 339 Rhipicephalus (Boophilus) microplus and 61 Haemaphysalis longicornis ticks were identified in Group 1 and Group 3. By molecular detection, 32 ‘Ca. M. haemobos’-positive amplicons, and 27 HoBi-like pestivirus-positive amplicons were amplified from goats in Group 1; meanwhile, 20 ‘Ca. M. haemobos’-positive amplicons, and 18 HoBi-like pestivirus-positive amplicons were amplified from foetuses. Statistical significance (P = 0.002) and association (OR=7.556) between the ‘Ca. M. haemobos’ PCR results of foetus and goat samples were observed, and statistical significance (P = 0.017) and association (OR=4.271) between the HoBi-like pestivirus PCR results of foetus and goat samples were observed. These significances and associations were also observed in Group 3. In addition, coinfections were detected in Group 1 and Group 3. ‘Ca. M. haemobos’ was detected in both tick species. Further serological results revealed that the frequency of HoBi-like pestivirus was 20.0 % (11/55) in Group 1 % and 30.6 % (11/36) in Group 3 in central China. No ticks, pathogens, or neutralizing antibodies were observed in Group 2 or Group 4. This is the first molecular evidence of ‘Ca. M. haemobos’ and HoBi-like pestivirus natural coinfections in goats and cows with abortion in China. HoBi-like pestivirus and ‘Ca. M. haemobos’ can be transferred from goats/cows to their foetuses.

Study of the allelic frequency of HSPA6 gene and its genotypes and their effect on some characteristics of milk production in Holstein cows

The study was conducted at the Taj Al-Nahrain cattle station in Al-Diwaniyah Governorate, which is 200 km from the center of Dhi Qar Governorate. For the period from 10/15/2021 to 10/3/2022, 49 Holstein cows imported from Germany were used in this experiment, all in the third production cycle. And the laboratory part of the study was conducted in the laboratory of the Marsh Research Center \ Dhi Qar University for the purpose of separating the genetic material from cow blood samples and conducting genetic analysis on it and extracting the possible genotypes HSPA6 heat shock protein gene. The animals in the station were fed according to the season and the year in terms of the availability of fodder and the percentage of costs involved in raising the cows, as well as on the general state of the productivity of the cows, but in general they are provided with green fodder, which includes jet, barley, corn stalks and a percentage of each of the dry fodder (hay and hay). ) and concentrate (barley, bran and cake) according to what is available from them, as well as limestone and table salt. Data on milk production were collected from the station records, where the milk production was recorded weekly for the morning and evening fenugreek by the station, and the milk components were analyzed in the Abu Dairy Company Gharib / Research and Development Division by Milk Analyzer. 3 ml of blood was collected from the udder vein of each cow by 5 ml medical syringes, suitable for the size of the animal’s vein, and placed in collection tubes containing an anticoagulant substance (K2 EDTA) and transported in a cooler box for safekeeping Freezing at -18°C until extraction time.
 From the results obtained from this study, we can conclude that the mutation detected in the studied segment of the 6HSPA gene was a type of missense mutation and worked to change the code of the amino acid valine to the code of the amino acid leucine. Three genotypes were shown: GG, GT, and TT. It was found that there was a clear commonality of cows carrying the GG genotype with a rate of 88%, followed by the GT genotype with 8%, while the TT genotype got the lowest percentage and amounted to 4%, and with allelic recurrence Its value is 0.92 for the G allele and 0.08 for the T . allele
 Groups of cows carrying the genotypes (GG, GT and TT) did not differ significantly among themselves in the characteristics of milk production and its components, but an arithmetic superiority between the three genotypes was observed and referred to in all the studied traits.

Serological, Molecular Prevalence and Genotyping of Coxiella burnetii in Dairy Cattle Herds in Northeastern Algeria.

In Algeria, data on the epidemiology of coxiellosis in cattle are still lacking. In this study, bulk tank milk (BTM) samples from 200 randomly selected dairy cattle herds from Setif province of Algeria were analyzed by an indirect enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Results highlighted that 37% (95% CI: 30.31–43.69%) and 9% (95% CI: 5.03–12.96%) of BTM samples contained Coxiella burnetii antibodies and DNA, respectively. Based on Cohen’s kappa coefficient, a very low agreement between the ELISA and PCR results was found (k = 0.0849) (95% CI: 0.00–0.189). For a second experiment, 186 whole blood samples of cows from farms with reproduction disorders were analyzed by molecular tools to detect C. burnetii. This study revealed an overall prevalence of 6.98% (95% CI: 3.32–10.65%). All positive samples determined by conventional PCR were analyzed by real-time quantitative PCR (qPCR). Eleven samples with cycle threshold (Ct) values lower than 35 were selected for genotyping by the multispacer sequence typing (MST) method. The MST12 genotype in BTM samples, the MST32 genotype and a new MST genotype (partial profile) in whole blood samples were identified. Obtained results have allowed us to better understand the epidemiology of bovine coxiellosis in the region of Setif.

PSIII-7 Relationship of Age and Genetics with the Methylation Profile of Beef Cattle

Abstract This study aimed to compare models for the prediction of cow age from DNA methylation profiles and estimate the heritability of the proportion of methylated sites (PM) and methylation status at each site (MS). Methylation data from blood samples of cows (n=136) were generated from the HorvathMammalMethylChip40 array that consists of 34,324 CpG sites that mapped to the bovine genome. Methylation status was determined by the distribution of the methylation values, with values above, within and below 2 standard deviations classified as methylated (2), intermediately methylated (1) and unmethylated (0), respectively. Principal component analysis (PCA) was applied to a (co)variance methylation status matrix. The first and second PC accounted for 25.65% and 9% of the total variance, respectively. Five Bayesian models (Bayesian ridge regression, BayesA, BayesB, BayesCπ and Bayesian LASSO) were implemented with the BGLR package in R. Bootstrapping validation (n=400) was used to evaluate the tested models, with 102 and 34 individuals in the training and validation sets, respectively. The correlation between the predicted and true age was high (r = 0.97 to 0.99). A BayesA model performed the best (r = 0.99, MSE = 0.11 and slope = 0.93), while Bayesian LASSO was the least accurate (r = 0.97, MSE = 0.26 and slope = 0.88). Heritability was estimated using GBLUP implemented in the BGLR package. The mean (SD) heritability estimate for PM was 0.46 ± 0.10 and the heritability of MS ranged from 0.18 to 0.73 (mean = 0.33). The 10% of sites with the highest heritability (343 sites; mean = 0.62) were located in exon (91), intron (84), intergenic (152), and promoter (16) regions. The largest number of these top sites (31) were located on chromosome 3 in genetic or intergenic regions close to transcription factor binding sites (i.e., FOXO6, ELAV4 and LMO4).

Monitoring of copper accumulation in water, soil, forage, and cows impacted by heavy automobiles in Sargodha, Pakistan.

The instant endeavor was undertaken to monitor copper (Cu) contents in water, soil, forage, and cow's blood impacted by heavy automobiles in Sahiwal town of district Sargodha, Pakistan. The samples were collected in triplicates with a total of 120 soil and water samples with corresponding forage samples. For the analysis of metal concentration in cows, 60 blood samples were collected from the cows feeding on these forages on selected sites. Metal contents were analyzed by atomic absorption spectrophotometry. The results showed that water samples contained mean values of Cu concentration ranged from 1.01 to 0.444 mg/kg at all sites. It was maximum at site 3 and minimum at site 6. The soil samples of all the forage fields showed Cu mean values concentration ranged from 1.94 to 0.286 mg/kg at all sites. It was maximum in Trifolium alexandrinum grown field at site 2, and minimum in Avena sativa at site 2. All the forage samples showed the mean value of Cu concentration ranged from 0.151 to 1.86 mg/kg at all sites. The concentration of Cu was maximum in Zea mays grown at site 5 and minimum in Trifolium alexandrinum at site 4. The cow blood samples showed the mean concentration of Cu ranged from 1.368 to 0.53 mg/kg at all sites. It was maximum at site 2 and minimum at site 6. Owing to the results of pollution index and transfer factors, metal content was found to be in permissible range in forages as well as animal samples.

Beta-Carotene concentration in blood serum of cows from herds with impaired fertility

Beta-Carotene concentration in blood serum of cows from herds with impaired fertility

PSIV-22 Validation of 26a and let7 microRNA as early pregnancy markers in single cow blood samples, at day 28 after embryo transfer

Abstract In this work we validated the previous scientific reports done in pooled samples by two different groups, over the miRNA expression in single blood from pregnant cows. Particularly we indagate the expression at day 28 after embryo transfer of let 7a, and 26a as well as 16 microRNAs (miRNA) family. By qPCR technique, we demonstrated that let 7a, and 26a increase maximum 2.1 and 2.6 fold change respectively in pregnant vs non-pregnant nulliparous heifers. This represents an exciting opportunity to use the differential expression of these miRNAs as early pregnancy markers and better understand the relationship with the cellular and molecular interaction between mother and embryo.

Identification of Polymorphisms of Gene CSN2 of B Casein in Greek Cow Breeds (Holstein) by Restriction Fragment Length Polymorphism

The research focused to detect and identify genetic polymorphisms in exon 7 of the β-casein CSN2 gene in blood samples from Greek Holstein cows. For this purpose, DNA was extracted from 120 blood samples of cows. The desired region of exon 7 was amplified by PCR, resulting in a 121 bp product. The PCR product was digested by restriction fragment length polymorphism (RFLP) method. The results suggest that the A1A2 genotype prevails over the others. Specifically, of the 120 cattle, 72 showed triple bands of 121 bp, 86 bp, and 35 bp indicating the A1A2 genotype. The 42 cattle showed a single band at 121 bp, indicating that they carried the A1A1 genotype. The remaining 6 showed only two bands of 86 and 35 bps, indicating that they carried the A2A2 genotype. In the total population of heterozygotes A1A2-0.60 were the most frequent, while homozygotes A2A2-0.06 were the least frequent ones. This suggests a slight superiority of allele A-0.65.

Transmission of Coxiella burnetii to Calves from Infected Cows

Background:Coxiella burnetii is the causative agent of a very important disease with zoonotic potential. Infected cows represent risk for spreading of infection to humans and to other animals on farm and also to their offspring. There is possibility for calves from infected cows to be infected nearly after parturition or during intrauterine life. Studies have shown that Coxiella burnetii initially infects the placenta and subsequent spread to the fetus may occur either by haematogenous or by the amniotic-oral route providing congenital infection. The main objective of the present study is to determine the presence of Coxiella burnetii genome in milk serum of infected cows and blood serum of calves. Materials, Methods & Results: A total of 200 blood serums from dairy cows were tested for presence of antibodies to Coxiella burnetii and nine of those were found positive. These animals compiled experimental group. From animals in experimental group milk samples during lactation, pregnancy and the postpartum period were collected. Samples were used for performing PCR test for determination of Coxiella burnetii presence in milk serum. On calving of each cow blood samples were taken from calves during first 24 hours after calving, from jugular vein. These blood samples were also used for PCR test to determine the presence of Coxiella burnetii. Milk serum analysis showed presence of Coxiella burnetii genome in serum, indicating on intermittent excretion. During lactation, the excretion of bacteria was greatest in the second stage when 80% of milk serum samples were positive for Coxiella burnetii. In the colostrums stage, there was a high percentage of Coxiella burnetii excretion through milk (50% of positive milk serum samples). The lowest percentage of excretion through milk was in the first stage of lactation. Analyzing blood serum samples from calves, taken on first day at calving using PCR method, all serums were positive for presence of Coxiella burnetii genome. Discussion:In animals, Coxiella burnetii is found in the reproductive system, both uterus and mammary glands, and may cause abortion or infertility. The high prevalence of Coxiella burnetii infection in dairy cattle with reproductive problems showed that these infected cattle play an important role in maintaining the infection and in disseminating of pathogenic agent to environment. The lowest percentage of excretion of Coxiella burnetii was in the first stage of lactation, amounting to 16.6%. With the transition to the next stage of lactation, a striking increase in the percentage of excretion was noticed. In the second stage it was 80%. In the third stage there was decrease to 40.6%. In colostrums stage percentage of excretion through milk was 50%. Similar results claims that excretion of Coxiella burnetii through milk starts after eight to twelve weeks of lactation in most cows. This period coincides with second stage of lactation as we divide it. Blood serums taken from calves were proven positive on Coxiella burnetii which indicates on intrauterine infection as described in the literature. Intrauterine infection takes place after placenta infection when bacteria penetrate the placenta and contaminate the amniotic fluid and gets aspirated or swallowed by fetus. Besides this,haematogenous spread can also occur.

Amelioration effects of Kaempferol on immune response following chronic intermittent cold-stress

Cold-stress causes disturbance of the homeostatic regulation of animals, and gradually impairs the immune and antioxidant functions of animals. Therefore, increasing the effectiveness of the immune response and antioxidant function are the most attractive strategies against cold-stress. Kaempferol (KPF) exerts both an anti-inflammatory and antioxidant pharmacological effect. However, nor much is known of the effects of KPF on providing protection from cold-induced intestinal oxidative damage and improving immunity. This study investigated the effects of KPF on immune factors and intestinal antioxidation in the blood of cold-stressed mice.KPF was solubilized in diluted saline before administration. The mice were randomly divided into 4 groups: (1) control, (2) cold-stress, (3) KPF 25 mg/kg, and (4) cinnamon (CAM) 30 mg/kg groups. Groups (2)–(4) were exposed to cold stress once a day for 7 days. Cold-stress was achieved by exposing the mice to a temperature of −15 °C and 70 ± 10% humidity for 60 min, once a day. The histopathological changes in the small intestine of the mice were analyzed. The T lymphocyte populations in blood were measured using flow cytometry. The level of SLC6a4, 5-HT3 and 5-HTT in small intestine tissue was assessed using RT-PCR analysis. Cow blood samples were obtained for the hematological analysis.Kaempferol (KPF) (25 mg/kg) regularized the intestinal antioxidant activity in the cold stress animals. KPF was able to significantly (P < .05) return intestinal SLC6a4, 5-HT3 and 5-HTT levels to normal after it had increased due to cold-stress. KPF treatment prevented the cold stress-induced decrease in blood CD4+T cells and decrease CD8+T cells levels in mice. Improved hematological profiles were additionally observed on treatment cows with KPF.KPF compared favorably with cinnamon in cold stress management, suggesting cold stress disturbs the anti-inflammatory effect of KPF. Thus, KPF contributes to suppress the activated pro-inflammatory cytokines, IL-9, IL-13, CD8+T and neurochemicals, and to increase anti-inflammatory cytokines and CD4+T levels.

NILAI GLUKOSA DARAH DAN TOTAL PROTEIN PLASMA PADA SAPI SIMPO YANG MENDERITA TREMATODIASIS DI PETERNAKAN RAKYAT DESA LABUHAN RATU KABUPATEN LAMPUNG TIMUR

The research aims to determine the value of blood glucose and plasma total protein in Simpo cattle suffering from trematodiasis. The research was conducted in December 2018 by taking blood samples of Simpo cattle in East Lampung. The research design used Completely Randomized Design (CRD) with 3 treatments and 4 replications from Simpo cow blood samples. The treatment used is P0: Simpo cattle which are not infested by worms; P1: Simpo cattle infested with 1 type of worm; and P2: Simpo cattle infested with 2 types of worms. Blood glucose value were analyzed in Biolab Indonesia Partner and total plasma were analyzed at Lampung Veterinary Center. Data from the observations were analyze using variance at the real level (P &lt;0,10) and continued with the Small Significant Difference test. The results of this study showed that Simpo cattle suffering from trematodiasis had significant effected (P &lt;0,10) on blood glucose values and had no significant effected (P&gt; 0,10) on total plasma protein. The highest number of blood glucose values at P0(65,00 mg / dL) and the lowest at P1 (56,75 mg / dL). Total plasma protein at P0 (7,20 g / dL), P1 (7,30 g / dL), and P2 (6,95 g / dL). &#x0D; Key word : Blood glucose, Total plasma protein, Simpo cattle, Trematodiasis, Traditional farmer

Identification on Osteopontin Promoter Gene Polymorphism and Post-thawing Quality in Dairy Bull Peranakan Friesian Holstein

This study aims to determine the existence of osteopontin promoter gene polymorphism in dairy bull Peranakan Holstein Friesian (PFH) and its relationship with the quality of PFH bull frozen semen. A total of 10 Holstein Friesian dairy cow blood samples were taken and then DNA extracted and amplified using SPP1F and SPP1R primers. The target band 306bp was detected in all samples and continued by sequencing to analyze the nucleotide bases. The results showed that sample with low quality frozen semen were deleted in the 10098 base and a transition (T-C) in bases 10054. The results indicated that this mutation site could be related to the trait susceptibility to frozen semen quality. Comprehensive studies are highly needed to address other parameters related to any abnormality in sperm during cryopreservation.

Some physiological and chemical properties of horse and cow blood samples under effect of some different types of anticoagulants

The purpose of the present study is to determine and compare the anticoagulants, Ethylene diamine tetra acetic acid, Sodium oxalate, Sodium citrate and heparin in blood samples of horse and bull, through estimation of (Hemoglobin, Packed Cell Volume and Plasma Platelets Count) and plasma physical properties (clarity and volume) and some chemical properties (pH and calcium ion concentration). Five Blood samples were obtained from five stallions and five bulls (20 ml/sample) and were divided as 5 ml in four test tubes containing Ethylene diamine tetra acetic acid, sodium oxalate, sodium citrate and heparin at the College of Veterinary Medicine, University of Diyala. Hemoglobin and packed cell volume were estimated immediately and then the blood samples were centrifuged for 15 min. at 3000 rpm to obtain plasma for evaluating the physical and chemical properties specified above. The horse blood samples pertaining data had no significant elevation in hemoglobin concentration and packed cell volume in Ethylene diamine tetra acetic acid group as compared to other groups, beside having no significant changes in pH between the four treated groups while there was a significant increase in plasma volume of oxalate and citrate as compared to Ethylene diamine tetra acetic acid and heparin, and there was a significant increase in plasma platelet count of Ethylene diamine tetra acetic acid group as compared to heparin. As well there was a significant decrease of calcium ion concentration in Ethylene diamine tetra acetic acid, oxalate and citrate as compared to heparin, while the results of bull blood samples revealed that the hemoglobin and Packed Cell Volume were significantly higher in heparin group than in the oxalate and citrate groups with non significant differences with Ethylene diamine tetra acetic acid group. The plasma obtained from different types of anticoagulants appeared with a high degree of clarity, and the volume of plasma had no significant increase in oxalate and citrate group as compared to other groups. The Plasma Platelet Count and calcium concentration significantly increased in heparin group as compared to other groups. In conclusion, the Ethylene diamine tetra acetic acid was more reliable for horse blood samples while the anticoagulant heparin was more reliable anticoagulant for bull blood samples.

Vaccinia Virus in Blood Samples of Humans, Domestic and Wild Mammals in Brazil.

Outbreaks of Vaccinia virus (VACV) affecting cattle and humans have been reported in Brazil in the last 15 years, but the origin of outbreaks remains unknown. Although VACV DNA have been already detected in mice (Mus musculus), opossums (Didelphis albiventris) and dogs during VACV zoonotic outbreaks, no transmission to cattle or humans from any of these were reported during Brazilian outbreaks. In this work, we assessed the PCR positivity to VACV in blood samples of cows and other domestic mammals, wild rodents and other wild mammals, and humans from areas with or without VACV infection reports. Our results show the detection of VACV DNA in blood samples of cows, horse and opossums, raising important questions about VACV spread.

Graphene-based multiplexed disposable electrochemical biosensor for rapid on-farm monitoring of NEFA and βHBA dairy biomarkers.

Elevated circulating concentrations of non-esterified fatty acids (NEFA) and beta hydroxy butyrate (βHBA) in biological fluids are recognized as critical biomarkers for early diagnosis of negative energy balance (NEB) in dairy cows. Herein, we report the development of a cost-effective, bio-friendly and electrochemically active dual screen printed electrode (SPE) sensor platform composed of electro-reduced graphene oxide nanosheets (E-rGO) modified with specific antibodies against NEFA and βHBA. The chemically synthesized graphene oxide (GO) was reduced directly on the screen printed electrode (SPE) surface via a green electrochemical approach without using toxic chemicals. The E-rGO was characterized using various analytical techniques, like XPS, SEM, TEM, AFM, UV-Vis, Raman spectroscopy, FTIR, and XRD, to get insight into its properties. Electrochemical analysis demonstrates that the E-rGO-modified SPE electrodes exhibit enhanced and durable redox properties as compared to the pristine graphite and GO electrodes. Target specificity is accomplished through immobilization of specific antibodies against NEFA and βHBA over the nanostructure-modified surface of the SPE, which only interacts with its counterpart NEFA and βHBA only. The antibodies retain their characteristic immuno-complex formation property upon immobilization and exhibit changes to amperometric signals upon interaction with various concentrations of NEFA and βHBA in standard, spiked blood and real clinical samples. The DPV signals resulted from the developed immunosensor platform exhibited a good correlation (R2∼ 0.99 for both NEFA and βHBA) for a wide range of target concentrations from 0.1 mM to 10 mM. The proposed immunosensor design not only provides a rapid analytical response time (≥1 min), but also simplicity in fabrication and instrumentation, which may provide a promising approach for on-farm diagnostics of ketosis and metabolic disorders associated with NEB.

Influence of Blood Sample Hemolysis on Hematological and Biochemical Parameters in Cows During Early Lactation

Summary The aim of this study is to investigate the influence of hemolysis degrees on the values of hematological and biochemical parameters in the blood samples of cows in early lactation. The study included a total of 45 blood samples of cows in early lactation. Out of the total number of samples analyzed, 15 indicated no signs of hemolysis, whereas 15 indicated moderate hemolysis and 15 severe hemolysis. The influence of hemolysis was examined by determining the bias (%) and by means of the Wilcoxon signed-rank test. The results showed that the hemolyzed samples had decreased values of the HCT, RBC, MCV, and increased values of the MCH, MCHC, and PLT. The total number of LEU showed a declining trend with increasing degrees of hemolysis. The changes in the HGB values were not detected despite the visual identification. The hemolysis degrees showed a significant influence on the values of biochemical parameters. The variations in biochemical parameters were less significant in the samples with moderate hemolysis compared to the samples with severe hemolysis. The most hemolysis-sensitive biochemical parameters were the AST, TBIL, TGC, and NEFA as great variations of their values were found even in moderate hemolysis. A slight deviation was noted in the BHB and total protein values. With respect to all the parameters examined, the Wilcoxon test showed significant differences in the samples with moderate and severe hemolysis in comparison with the control group. In conclusion, hemolysis significantly influenced the values of hematological and biochemical parameters in the blood samples analyzed.

Use of serology and real time PCR to control an outbreak of bovine brucellosis at a dairy cattle farm in the Nile Delta region, Egypt.

BackgroundBovine brucellosis remains one of the most prevalent zoonotic infections affecting dairy cattle in developing countries where the applied control programs often fail. We analyzed the epidemiologic pattern of bovine brucellosis in a dairy cattle herd that showed several cases of abortions after regular vaccination with RB51 (B. abortus vaccine). In 2013 thirty dairy cows, from a Holstein-Friesian dairy herd with a population of 600 cattle, aborted five months post vaccination by a regular RB51 vaccine. Blood samples were drawn from milking cows and growing heifers, as well as heifers and cows pregnant up to 6 months. These samples were collected in June 2013 (n = 257) and May 2014 (n = 263) and were tested by real time (rt)-PCR as well as serological tests, in particular Rose Bengal Test (RBT), Enzyme-Linked Immunosorbent Assays (ELISA) and Fluorescence Polarization Assay. Tissue specimens were also collected from an aborted fetus and cultured. Isolates were subjected to bacteriological typing tests at the genus and species levels.ResultsFive months post vaccination with RB51 vaccine, Brucella (B.) DNA was detected in blood samples of cows by rt-PCR. The serological tests also revealed the spread of Brucella field strains within the herd in 2013. Four Brucella isolates were recovered from specimens collected from the aborted fetus. These isolates were typed as follows: one B. abortus RB51 vaccine strain and three isolates of B. abortus field strain. The seropositive cows with positive rt-PCR might indicate an infection by the Brucella field strain; while the positive rt-PCR results from seronegative animals may either be due to circulating RB51 vaccine DNA in vaccinated animals or to circulating field strain in infected animals before seroconversion.ConclusionThe results herein suggest that PCR can be a good supplementary tool in an outbreak situation, if an assay is available that can differentiate vaccine and field strains with a high analytical sensitivity. We recommend using RBT and ELISA in parallel in outbreak situations, to identify as many infected animals as possible during the initial screenings. This test procedure should be repeated for at least three successive negative tests, with one month interval.