Epigenetic states of genes controlling immune responsiveness in bovine chronic mastitis

Abstract Mastitis is a common disease in dairy cows, causing substantial economic losses. The leading cause of elevated milk somatic cell count (SCC), which is the best indicator for detecting mastitis, is the invasion of pathogens. A major pathogen responsible for bovine mastitis is Staphylococcus aureus, a member of the coagulase-positive staphylococci. Some strains of coagulase-negative staphylococci can also be a cause of clinical or subclinical mastitis. Our study used bisulfite sequencing PCR (BSP) to detect the methylation status of nine candidate genes (CCL2, HCK, F11R, CD8A, PDIA3, LGMN, HSPA1A, IL18 and NFKBIA). We investigated the mechanisms associated with overexpression of these genes, in the mammary gland secretory tissue of cows diagnosed with mastitis and infected with coagulase-positive or coagulase-negative staphylococci. The results showed no changes at the DNA methylation level between the mastitis (CoPS and CoNS) and control groups (H), except for in the HCK region, where the observed differences between the CoPS and H groups were statistically significant. The low methylation level of the CpG sequence seems not to correspond to the previously observed increased activity of these genes, suggesting that mechanisms other than DNA methylation may control mRNA expression at the analyzed loci.

Download Full-text

Therapeutic Effect of Nisin Z on Subclinical Mastitis in Lactating Cows

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00629-07 ◽

2007 ◽

Vol 51 (9) ◽

pp. 3131-3135 ◽

Cited By ~ 73

Author(s):

Junqiang Wu ◽

Songhua Hu ◽

Liting Cao

Keyword(s):

Subclinical Mastitis ◽

Treated Group ◽

Economic Losses ◽

Control Group ◽

Therapeutic Effects ◽

Coagulase Negative Staphylococci ◽

Milk Samples ◽

Lactating Cows ◽

Nisin Z ◽

And Control

ABSTRACT Bovine subclinical mastitis is an inflammation of the mammary gland caused by bacterial intramammary infection, accounting for large economic losses. Treatment of subclinical mastitis is not suggested for lactating cows due to the risk of milk contamination. The objectives of this study were to evaluate an antimicrobial peptide, nisin, in the treatment of subclinical mastitis in lactating cows. A total of 90 lactating Holstein cows with subclinical mastitis were randomly divided into nisin-treated (n = 46) and control (n = 44) groups. In the nisin-treated group, cows received an intramammary infusion of nisin at a dose of 2,500,000 IU once daily for 3 days while the control cows received no treatment. Milk samples were collected from the affected mammary quarters before treatment and 1 and 2 weeks after treatment for analyses of bacteria, somatic cells, and N-acetyl-β-d-glucosaminidase (NAGase). Results indicated that nisin therapy had bacteriological cure rates of 90.1% for Streptococcus agalactiae (10 of 11), 50% for Staphylococcus aureus (7 of 14), 58.8% for coagulase-negative staphylococci (7 of 17), and 65.2% for all cases (30 of 46). Meanwhile, only 15.9% (7 of 44) of untreated cows spontaneously recovered. NAGase activity in milk samples and the number of mammary quarters with a milk somatic cell count of ≥500,000/ml were significantly decreased after nisin treatment while no significant changes took place in the control group. Because of its therapeutic effects on bovine subclinical mastitis, as well as its safety in humans, nisin deserves further study to clarify its effects on mastitis caused by different pathogens.

Download Full-text

Microbiota of Cow’s Milk with Udder Pathologies

Microorganisms ◽

10.3390/microorganisms9091974 ◽

2021 ◽

Vol 9 (9) ◽

pp. 1974

Author(s):

Mariya V. Gryaznova ◽

Mikhail Y. Syromyatnikov ◽

Yulia D. Dvoretskaya ◽

Sergey A. Solodskikh ◽

Nikolay T. Klimov ◽

...

Keyword(s):

Bovine Mastitis ◽

Subclinical Mastitis ◽

Sensu Stricto ◽

Relative Increase ◽

Cow’S Milk ◽

Economic Losses ◽

Common Disease ◽

Rrna Gene ◽

Cow's Milk

Mastitis is the most common disease for cattle, causing great economic losses for the global dairy industry. Recent studies indicate the multi-agent and microbiome diversity of this disease. To understand the nature of mastitis and investigate the role of the microbiome in the development of pathologies in the udder of bovines, we performed NGS sequencing of the 16S rRNA gene of cow’s milk with pathologies of the udder. The obtained data show a significant increase in the Cutibacterium, Blautia, Clostridium sensu stricto 2, Staphylococcus, Streptococcus and Microbacterium genera for groups of cows with udder pathologies. Increasing relative abundance of the Staphylococcus and Streptococcus genera was associated with subclinical mastitis. Our data show that a relative increase in abundance of the Staphylococcus and Microbacterium genera may be an early sign of infection. We have shown, for the first time, an increase in the Colidextribacter, Paeniclostridium and Turicibacter genera in groups of cows with mastitis. These results expand our understanding of the role of the microbiome in the development of bovine mastitis.

Download Full-text

DNA methylation of insulin-like growth factor 2 and H19 cluster in cord blood and prenatal air pollution exposure to fine particulate matter

Environmental Health ◽

10.1186/s12940-020-00677-9 ◽

2020 ◽

Vol 19 (1) ◽

Author(s):

Congrong Wang ◽

Michelle Plusquin ◽

Akram Ghantous ◽

Zdenko Herceg ◽

Rossella Alfano ◽

...

Keyword(s):

Dna Methylation ◽

Particulate Matter ◽

Growth Factor ◽

Cord Blood ◽

Gene Cluster ◽

Methylation Level ◽

Fine Particulate Matter ◽

Methylation Status ◽

Insulin Like Growth Factor ◽

Fine Particulate

Abstract Background The IGF2 (insulin-like growth factor 2) and H19 gene cluster plays an important role during pregnancy as it promotes both foetal and placental growth. We investigated the association between cord blood DNA methylation status of the IGF2/H19 gene cluster and maternal fine particulate matter exposure during fetal life. To the best of our knowledge, this is the first study investigating the association between prenatal PM2.5 exposure and newborn DNA methylation of the IGF2/H19. Methods Cord blood DNA methylation status of IGF2/H19 cluster was measured in 189 mother-newborn pairs from the ENVIRONAGE birth cohort (Flanders, Belgium). We assessed the sex-specific association between residential PM2.5 exposure during pregnancy and the methylation level of CpG loci mapping to the IGF2/H19 cluster, and identified prenatal vulnerability by investigating susceptible time windows of exposure. We also addressed the biological functionality of DNA methylation level in the gene cluster. Results Prenatal PM2.5 exposure was found to have genetic region-specific significant association with IGF2 and H19 during specific gestational weeks. The association was found to be sex-specific in both gene regions. Functionality of the DNA methylation was annotated by the association to fetal growth and cellular pathways. Conclusions The results of our study provided evidence that prenatal PM2.5 exposure is associated with DNA methylation in newborns’ IGF2/H19. The consequences within the context of fetal development of future phenotyping should be addressed.

Download Full-text

Repeats as global DNA methylation marker in bovine preimplantation embryos

Czech Journal of Animal Science ◽

10.17221/29/2016-cjas ◽

2017 ◽

Vol 62 (No. 2) ◽

pp. 43-50 ◽

Cited By ~ 1

Author(s):

W. Li ◽

A. Van Soom ◽

L. Peelman

Keyword(s):

Dna Methylation ◽

Mutation Rate ◽

Methylation Level ◽

Methylation Status ◽

Repetitive Sequences ◽

Cell Types ◽

Blastocyst Stage ◽

Repeat Sequence ◽

Preimplantation Embryos ◽

Global Dna Methylation

DNA methylation undergoes dynamic changes and is a crucial part of the epigenetic regulation during mammalian early development. To determine the DNA methylation levels in bovine embryos, we applied a bisulfite sequencing based method aimed at repetitive sequences including three retrotransposons (L1_BT, BovB, and ERV1-1-I_BT) and Satellite I. A more accurate estimate of the global DNA methylation level compared to previous methods using only one repeat sequence, like Alu, could be made by calculation of the weighted arithmetic mean of multiple repetitive sequences, considering the copy number of each repetitive sequence. Satellite I and L1_BT showed significant methylation reduction at the blastocyst stage, while BovB and ERV1-1-I_BT showed no difference. The mean methylation level of the repetitive sequences during preimplantation development was the lowest at the blastocyst stage. No methylation difference was found between embryos cultured in 5% and 20% O<sub>2</sub>. Because mutations of CpGs negatively influence the calculation accuracy, we checked the mutation rate of the sequenced CpG sites. Satellite I and L1_BT showed a relatively low mutation rate (1.92 and 3.72% respectively) while that of ERV1-1-I_BT and BovB was higher (11.95 and 24% respectively). Therefore we suggest using a combination of repeats with low mutation rate, taking into account the proportion of each sequence, as a relatively quick marker for the global DNA methylation status of preimplantation stages and possibly also for other cell types.

Download Full-text

Investigating skewness to understand gene expression heterogeneity in large patient cohorts

BMC Bioinformatics ◽

10.1186/s12859-019-3252-0 ◽

2019 ◽

Vol 20 (S24) ◽

Cited By ~ 2

Author(s):

Benjamin V. Church ◽

Henry T. Williams ◽

Jessica C. Mar

Keyword(s):

Dna Methylation ◽

Methylation Status ◽

Microarray Technology ◽

Statistical Measure ◽

Large Patient ◽

Technology Platforms ◽

Gene Expression Heterogeneity ◽

Cancer Genome Atlas ◽

Or Genes ◽

And Control

Abstract Background Skewness is an under-utilized statistical measure that captures the degree of asymmetry in the distribution of any dataset. This study applied a new metric based on skewness to identify regulators or genes that have outlier expression in large patient cohorts. Results We investigated whether specific patterns of skewed expression were related to the enrichment of biological pathways or genomic properties like DNA methylation status. Our study used publicly available datasets that were generated using both RNA-sequencing and microarray technology platforms. For comparison, the datasets selected for this study also included different samples derived from control donors and cancer patients. When comparing the shift in expression skewness between cancer and control datasets, we observed an enrichment of pathways related to the immune function that reflects an increase towards positive skewness in the cancer relative to control datasets. A significant correlation was also detected between expression skewness and the top 500 genes corresponding to the most significant differential DNA methylation occurring in the promotor regions for four Cancer Genome Atlas cancer cohorts. Conclusions Our results indicate that expression skewness can reveal new insights into transcription based on outlier and asymmetrical behaviour in large patient cohorts.

Download Full-text

Prevalence, Etiology, and Risk Factors of Mastitis in Dairy Cattle in Embu and Kajiado Counties, Kenya

Veterinary Medicine International ◽

10.1155/2020/8831172 ◽

2020 ◽

Vol 2020 ◽

pp. 1-12

Author(s):

Christine M. Mbindyo ◽

George C. Gitao ◽

Charles M. Mulei

Keyword(s):

Management Practices ◽

Health Hazard ◽

Previous History ◽

Bovine Mastitis ◽

Subclinical Mastitis ◽

Control Measures ◽

Economic Losses ◽

Isolation And Identification ◽

Cross Sectional ◽

Milk Samples

Bovine mastitis continues to be a leading cause of heavy economic losses in the dairy industry and a public health hazard globally. This cross-sectional study investigated the prevalence, etiologies of clinical and subclinical mastitis, and associated predisposing factors in Embu and Kajiado counties in Kenya. A semistructured questionnaire was administered to 154 smallholder dairy farmers to collect data on management practices, animal factors, and disease history. A total of 395 dairy cows were initially screened for subclinical mastitis using the California mastitis test (CMT), and milk samples were aseptically collected. Both CMT positive and CMT negative samples were analyzed using conventional bacteriological isolation and identification procedures. In the present study, the overall prevalence of mastitis based on CMT and clinical examination was 80% (316/395), out of which 6.8% (27/395) was clinical mastitis, while 73.1% (289/395) was subclinical mastitis. Based on culture, the overall prevalence of clinical and subclinical mastitis was 51.6% (815/1580), 74.4% (294/395), and 76.6% (118/154) at the quarter, cow, and farm level, respectively. From the 1574 milk samples analyzed by cultured, 1016 bacteria were yielded. The predominant bacteria were coagulase-negative Staphylococcus (CNS), 42.8% (435/1016), and in decreasing order, Streptococcus species, 22.2% (226/1016), Staphylococcus aureus, 15.7% (160/1016), and Pseudomonas aeruginosa, 5.1% (52/1016), and the least was Enterobacter species, 0.7% (7/1016), while 23.7% of the sample yielded no bacterial growth. Risk factor analysis revealed that milking mastitic cows last (p=0.002), using a clean udder drying towel for each cow (p=0.033) and previous history of mastitis (p=0.046) were significantly associated with presence of mastitis. The current study has shown a relatively high prevalence of subclinical mastitis with CNS as predominant bacteria. Therefore, control measures are urgently warranted. Management factors such as milking mastitic cows last, using a clean towel for udder drying for each cow, and culling mastitic cows should be considered and included in the Kenyan mastitis control programs.

Download Full-text

Standardization of DNA amount for bisulfite conversion for analyzing the methylation status of LINE-1 in lung cancer

PLoS ONE ◽

10.1371/journal.pone.0256254 ◽

2021 ◽

Vol 16 (8) ◽

pp. e0256254

Author(s):

Duong Anh Thuy Pham ◽

Son Duc Le ◽

Trang Mai Doan ◽

Phuong Thu Luu ◽

Uyen Quynh Nguyen ◽

...

Keyword(s):

Lung Cancer ◽

Dna Methylation ◽

Methylation Level ◽

Genomic Dna ◽

Methylation Status ◽

Cancer Tissue ◽

Nucleotide Position ◽

Bisulfite Conversion ◽

Precise Location ◽

Significant Variance

Highly methylated Long Interspersed Nucleotide Elements 1 (LINE-1) constitute approximately 20% of the human genome, thus serving as a surrogate marker of global genomic DNA methylation. To date, there is still lacking a consensus about the precise location in LINE-1 promoter and its methylation threshold value, making challenging the use of LINE-1 methylation as a diagnostic, prognostic markers in cancer. This study reports on a technical standardization of bisulfite-based DNA methylation analysis, which ensures the complete bisulfite conversion of repeated LINE-1 sequences, thus allowing accurate LINE-1 methylation value. In addition, the study also indicated the precise location in LINE-1 promoter of which significant variance in methylation level makes LINE-1 methylation as a potential diagnostic biomarker for lung cancer. A serial concentration of 5-50-500 ng of DNA from 275 formalin-fixed paraffin-embedded lung tissues were converted by bisulfite; methylation level of two local regions (at nucleotide position 300–368 as LINE-1.1 and 368–460 as LINE-1.2) in LINE-1 promoter was measured by real time PCR. The use of 5 ng of genomic DNA but no more allowed to detect LINE-1 hypomethylation in lung cancer tissue (14.34% versus 16.69% in non-cancerous lung diseases for LINE-1.1, p < 0.0001, and 30.28% versus 32.35% for LINE-1.2, p < 0.05). Our study thus highlighted the optimal and primordial concentration less than 5 ng of genomic DNA guarantees the complete LINE-1 bisulfite conversion, and significant variance in methylation level of the LINE-1 sequence position from 300 to 368 allowed to discriminate lung cancer from non-cancer samples.

Download Full-text

DNA Methylation Changes and Its Associated Genes in Mulberry (Morus alba L.) Yu-711 Response to Drought Stress Using MethylRAD Sequencing

Plants ◽

10.3390/plants11020190 ◽

2022 ◽

Vol 11 (2) ◽

pp. 190

Author(s):

Michael Ackah ◽

Liangliang Guo ◽

Shaocong Li ◽

Xin Jin ◽

Charles Asakiya ◽

...

Keyword(s):

Dna Methylation ◽

Gene Regulation ◽

Drought Stress ◽

Methylation Status ◽

Enrichment Analysis ◽

Plant Genome ◽

Pcr Analysis ◽

Kegg Pathways ◽

Wide Range ◽

And Control

Drought stress remains one of the most detrimental environmental cues affecting plant growth and survival. In this work, the DNA methylome changes in mulberry leaves under drought stress (EG) and control (CK) and their impact on gene regulation were investigated by MethylRAD sequencing. The results show 138,464 (37.37%) and 56,241 (28.81%) methylation at the CG and CWG sites (W = A or T), respectively, in the mulberry genome between drought stress and control. The distribution of the methylome was prevalent in the intergenic, exonic, intronic and downstream regions of the mulberry plant genome. In addition, we discovered 170 DMGs (129 in CG sites and 41 in CWG sites) and 581 DMS (413 in CG sites and 168 in CWG sites). Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis indicates that phenylpropanoid biosynthesis, spliceosome, amino acid biosynthesis, carbon metabolism, RNA transport, plant hormone, signal transduction pathways, and quorum sensing play a crucial role in mulberry response to drought stress. Furthermore, the qRT-PCR analysis indicates that the selected 23 genes enriched in the KEGG pathways are differentially expressed, and 86.96% of the genes share downregulated methylation and 13.04% share upregulation methylation status, indicating the complex link between DNA methylation and gene regulation. This study serves as fundamentals in discovering the epigenomic status and the pathways that will significantly enhance mulberry breeding for adaptation to a wide range of environments.

Download Full-text

Molecular identification of coagulase-negative staphylococci isolated from bovine mastitis and detection of β-lactam resistance

The Journal of Infection in Developing Countries ◽

10.3855/jidc.5871 ◽

2015 ◽

Vol 9 (09) ◽

pp. 1022-1027 ◽

Cited By ~ 5

Author(s):

Mariela Elizabeth Srednik ◽

Mario Andres Grieben ◽

Adriana Bentancor ◽

Elida Raquel Gentilini

Keyword(s):

Fragment Length Polymorphism ◽

Meca Gene ◽

Bovine Mastitis ◽

Economic Loss ◽

Economic Losses ◽

Dairy Herds ◽

Coagulase Negative Staphylococci ◽

Milk Samples ◽

Gap Gene ◽

Mastitis Pathogens

Introduction: Bovine mastitis is a frequent cause of economic loss in dairy herds. Coagulase-negative staphylococci (CoNS) are increasing in importance as causeof bovine intramammary infection (IMI) throughout the world in recent years. CoNShave been isolated from milk samples collected from cows with clinical andsubclinical mastitis in several countries. Identification of mastitis pathogensis important when selection appropriate antimicrobial therapy.Methodology: A total of 93 strains of Staphylococcusspp isolated from bovine clinical andsubclinical mastitis in Argentina during 2010-2013 were identified by Restriction Fragment Length Polymorphism (RFLP)-PCR using the gap gene. The isolates were tested by PCR for the presence of blaZ gene and mecA gene and were tested by disk diffusion for the susceptibilityto penicillin and cefoxitin. Results: The most common CoNS species was S.chromogenes 46.2% (43/93), followed by S. devriesei 11.8% (11/93) and S. haemolyticus 9.7% (9/93). The blaZ gene was detected in 19 (20.4%) but only 16 (17.2%) isolates were resistant to penicillin; the mecA was detected in6 (6.5%) isolates but only 4 (4.3) were resistant to cefoxitin. The 6 mecA-positive isolates showed oxaxillinMICs ≥ 0.5 μg/ml. Discussion: CoNSare important minor mastitis pathogens and can be the cause of substantial economic losses. Despite the low resistance to PEN in Argentina, the presenceof MR isolates found in this study emphasize the importance of identificationof CoNS when an IMI is present because of the potentially risk of lateraltransfer of resistance genes between staphylococcal species.

Download Full-text

Clinical and subclinical bovine mastitis in area of Kilkis

Journal of the Hellenic Veterinary Medical Society ◽

10.12681/jhvms.15791 ◽

2018 ◽

Vol 48 (1) ◽

pp. 32 ◽

Cited By ~ 1

Author(s):

N. ILIADIS (Ν. ΗΛΙΑΔΗΣ) ◽

E. N. PETRIDOU (Ε.N.ΠΕΤΡΙΔΟΥ) ◽

A. FOUKOS (Α. ΦΟΥΚΟΣ)

Keyword(s):

Bovine Mastitis ◽

Subclinical Mastitis ◽

Sensitivity Test ◽

Coagulase Negative Staphylococci ◽

Milk Samples ◽

Microbiological Examination

In this study 216 milk samples from equal number of quarters were examined. The results from microbiological examination and the performed California test revealed that 82 samples (38%) were collected from quartres with subclinical mastitis. Coagulase-negative staphylococci were the most frequent pathogens isolated from the examined milk samples, representing 47.5% of the total isolated strains. Sensitivity test of the strains to certain antimicrobials showed higher performance of gentamycine (90.9% of the cases). On a descending sequence cephalotin (75%), oxalinique acid (63.6%) and ampicilline (54.2%) were the antimicrobials to which sensitivity of the strains was higher than 50%. On the contrary, tetracycline, tincomycine, erythromycine and penicilline G performance was lower than 50%.

Download Full-text