Dairy Cattle Mammary Health—Reducing Mammary Disease

Mastitis is the most common production disease in the dairy sector worldwide, its incidence being associated with both cows’ exposure to bacteria and the cows’ genetic make-up for resistance to pathogens. The objective of our study was to analyse 89 missense SNPs belonging to six genes (CXCR2, CXCL8, TLR4, BRCA1, LTF, BOLA-DRB3), which were found to be associated with genetic resistance or susceptibility to mastitis. A total of 298 cattle (250 Romanian Spotted and 48 Romanian Brown) were genotyped by Kompetitive Allele Specific PCR (KASP) and a chi-squared test was used for genetic association studies with clinical mastitis. A total of 35 SNPs (39.3%) among the selected 89 SNPs were successfully genotyped, of which 31 markers were monomorphic. The polymorphic markers were found in two genes: TLR4 (rs460053411) and BOLA-DRB3 (rs42309897, rs208816121, rs110124025). The polymorphic SNPs with MAF > 5% and call rates > 95% were used for the association study. The results showed that rs110124025 in the BOLA-DRB3 gene was significantly associated with mastitis prevalence (p ≤ 0.05) in both investigated breeds. Current results show that the SNP rs110124025 in the BOLA-DRB3 gene can be used as a candidate genetic marker in selection for mastitis resistance in Romanian dairy cattle. Full article

Bovine mastitis is an inflammation of the mammary gland in response to invasion by opportunistic agents. Due to the high economic importance of dairy production and the complexity related to animal health, the objective of this work was to identify and evaluate the antibacterial resistance profile of samples of mastitis milk, milking hand and milking equipment from small rural dairy farms belonging to the northwest region of the state of Paraná, Brazil. Five small, non-technical dairy farms in the municipalities of Boa Esperança, Juranda and Tapejara, all belonging to the northwest region of the state of Paraná, Brazil, were selected. The properties had Holstein and/or crossbred herds, carried out a bucket-by-foot milking system and all had the presence of animals with subclinical mastitis confirmed by the California Mastitis Test. Samples of sterile swabs from the milking insufflators, the milking hand and milk samples were collected—and later, isolation tests and phenotypic characterization of the samples, sensitivity tests to antimicrobials and phenotypic tests for the detection of beta-producing strains were performed with extended-spectrum beta-lactamase (ESBL), molecular identification of Staphylococcus aureus isolates and mecA gene research. Of the 199 samples collected from the 15 selected properties in the municipalities of Boa Esperança, Tapejara and Juranda, 72 (36.20%) were classified as multiresistant. Isolated from milkers’ hands and milking machines, which phenotypically produce extended-spectrum beta-lactamase (ESBL), the presence of the mecA gene was also observed in 11 isolates of Staphylococcus spp. of milk samples, machines and milking hands. Mastitis can be spread to the herd through the milking process by the milkers’ instruments and hands, and adequate management measures can prevent its transmission and the conscious use of antibiotics decreases the prevalence of multidrug-resistant pathogens. In this work, different pathogenic bacteria were detected in mastitic milk, milking equipment and milking hand with a high percentage (36.20%) of isolates classified as multidrug resistant. In addition, the presence phenotypically (ESBL) and molecularly (mecA gene) of isolates carrying resistance genes was also verified. These results directly reflect on the health of the animals, the health of the workers and the health of the respective environment, which can enable the continuity of the propagation of the etiological agents involved in the mastitis infection. The awareness of producers and workers on these properties about the disease, transmission, sanitary aspects and adequate management and treatment are essential for improving milk production and production efficiency. Full article

Mastitis is a major cause of antimicrobial treatments either during lactation or at drying off. From a One Health perspective, there should be a balance between the risk of IMI that may impair cow health and welfare and the reduction of antimicrobial usage to decrease antimicrobial resistance, as may happen when applying selective dry-cow therapy. This reduction may be achieved by an early and accurate diagnosis followed by prudent and rationale therapeutical protocols. This study aims to assess the accuracy of PLCC (neutrophils + lymphocyte count/mL) in identifying cows at risk of having IMI due to major pathogens (S.aureus, Str.agalactiae, Str.uberis, and Str.dysgalactiae), and to simulate the impact of this early diagnosis on the potential number of treatments using a decision-tree model. The results of this study showed that PLCC had an overall accuracy of 77.6%. The results of the decision-tree model based on data from the 12 participating herds, with an overall prevalence of major pathogens of 1.5%, showed a potential decrease in the number of treatments of about 30% (from 3.4% to 2.5%) when PLCC in early lactation (days 5–16) was used to identify cows at risk for major pathogens compared with using SCC at the first milk test (days 17–43). The study confirmed that it is possible to improve animal health and reduce the risk of antimicrobial use through early IMI detection based on PLCC and applying a rationale and prudent antimicrobial protocol. Full article

Extracellular vesicles (EVs) are membranous vesicles found in biological fluids with essential functions. However, milk-derived EV proteins from clinical mastitis (CM) and subclinical mastitis (SM) cows have yet to be studied in detail. In this study, milk-derived EVs of CM, SM, and Healthy cows were extracted using a combination of acetic acid/ultracentrifugation and density gradient ultracentrifugation and analyzed using a shotgun proteomic by data-independent acquisition mode. A total of 1253 milk exosome proteins were identified and quantified. Differently enriched (DE) proteins were identified as given a Benjamini–Hochberg adjusted p < 0.05 and a fold change of at least 2. There were 53 and 1 DE proteins in milk-derived EVs from CM and SM cows compared with healthy cows. Protein S100-A9, Protein S100-A8, Chitinase-3-like protein 1, Haptoglobin, Integrin beta-2, and Chloride intracellular channel protein 1 were more abundant in the CM group (adjusted p < 0.05). Still, their enrichment in the SM group was not significant as in the Healthy group. The enrichment of DE proteins between CM and Healthy group was consistent with elevated GO (Gene Ontology) processes—defense response, defense response to Gram-positive bacterium, granulocyte chemotaxis also contributed to Reactome pathways—neutrophil degranulation, innate immune system, and antimicrobial peptides in the CM group. These results provide essential information on mastitis-associated proteins in milk-derived EVs and indicate the biological functions of milk-derived EVs proteins require further elucidation. Full article

Milk production loss due to mastitis in dairy herds is economically important. Before estimating the economic impacts of mastitis, it is crucial to quantify the association between mastitis and milk production. The objective of this study was to estimate the association between somatic cell count (SCC, as an indicator of intramammary infection due to mastitis) and milk production for dairy cows in Lombardy, Italy. The test-day (TD) records data of 3816 dairy herds located in three different geographical areas of Lombardy from January 2016 to December 2018 were used. After data editing, the final dataset comprised 10,445,464 TD records from 2970 farms and 826,831 cows. The analysis was carried out by using a mixed-effects model with six fixed effects (geographical Area, Breed, Days in Milk, Parity, Season and Year) and nested random effects for each cow and herd. The results confirmed that the SCC had a negative association with milk production. On average, this study found that any two-fold increase of SCC resulted in a milk production loss of 0.830 (95% CI: −0.832, −0.828) kg/cow/day in the whole of Lombardy. These results can be used for economic calculations on the costs of mastitis. Full article

Dairy cattle with a high milk yield are susceptible to many infectious diseases, such as mastitis. Subclinical mastitis (SCM) is the most prevalent form of mastitis that predominantly affects animal health, and causes adverse effects on the quality and quantity of milk. In dairy animals, subclinical mastitis often remains undetected, as no gross changes in udder characteristics are visible. In the present study, 135 Holstein Friesian dairy animals were selected and screened as healthy (n = 25) and mastitic (n = 110) based on diagnostic tests such as the California mastitis test, pH, electrical conductivity, and somatic cell count. In this study, the somatic cell count was used as a gold-standard test in differentiating subclinical mastitis animals from healthy ones. The present study was carried out to study polymorphisms in the bovine transferrin gene in cows (with subclinical mastitis and healthy). For the early detection of resistant/or susceptible animals, a useful marker could be provided by the detection of single-nucleotide polymorphisms (SNPs) in the transferrin gene, which are often associated with mammary innate immune response. The sequencing results revealed three nucleotide substitutions: two transversions (230 A > C, 231 C > A) and one transition (294 A > G) in susceptible cows as compared to disease-free subjects. The nucleotide variations at position 230 (GAC > GCA) and 231 (GAC > GCA) were nonsynonymous, and corresponded to an amino acid change from aspartic acid to alanine; whereas at position 294 (GAA > GAG), the mutation was synonymous. In the present study, many in silico tools were taken into consideration to determine the effect of SNPs on protein structure and function. The PROVEAN tool found the amino acid substitution to be neutral and deleterious. PolyPhen-2 revealed the amino acid variations at positions 320 and 321 to most likely be damaging; and at the 341 position, the variations were benign. The I-Mutant and MUpro tools found that the protein stability decreased for nonsynonymous variations. The SIFT tool revealed the protein function was likely to be affected in nonsynonymous variations, with no change in the case of synonymous ones. Phylogenetic analysis of the bovine transferrin gene revealed a close relation of the CA allele with the Bos taurus transferrin, while the G allele was closely related to a cross of Bos indicus × Bos taurus serotransferrins, followed by the Bison bison transferrin. The least relation was shown by both alleles to Capra hircus, Ovis aries, and Bubalus bubalis. Full article

Mastitis, which affects milk quality and yield, is one of the most common diseases in dairy cows, causing large economic losses. Cow mastitis is classified into clinical and subclinical types. Subclinical mastitis presents without obvious lesions in the udder or noticeable change in milk samples, indicating persistent chronic infection that is difficult to detect and treat. Therefore, finding specific biomarkers is of great significance for the early diagnosis and treatment of subclinical mastitis. As mediators of intercellular communication, exosomes have been shown to be extensively involved in various physiological and pathological processes in the body. Exosomes in milk, blood, and cell supernatant can carry stable cell source-specific nucleic acids, proteins, and metabolites. Hence, exosomes show great application prospects for early diagnosis, targeted therapy, and disease mechanism analysis. In this review, we summarize the biogenesis, biological functions, and methods of isolating and identifying exosomes and review the current status of exosome research related to mastitis. Finally, in view of the application of exosomes to diagnose, treat, and perform disease mechanism analysis in mastitis, deficiencies in recent research on mastitis exosomes are described, and the direction of future exosome research efforts in mastitis is proposed. Full article

The objectives of this paper were (i) to perform a systematic review of the literature over the last 21 yr and (ii) to evaluate the efficacy of selective dry cow treatment (SDCT) vs. blanket dry cow treatment (BDCT) in dairy cows regarding the risk of intramammary infection (IMI) after calving, new IMI risk after calving, cure risk during the dry period, and a reduction in antibiotic use at drying-off by meta-analysis. The systematic search was carried out using the databases PubMed, CAB Direct, and ScienceDirect. A meta-analytical assessment was performed for each outcome of interest using random-effects models, and the relative risk (RR) for IMI and cure or the pooled proportion for antibiotic use was calculated. The final number of included studies was n = 3 for IMI risk after calving and n = 5 for new IMI risk after calving, cure risk during the dry period, and antibiotic use. The RR levels for IMI (RR, 95% confidence interval [CI]: 1.02, 0.94–1.11; p = 0.592), new IMI (RR, 95% CI: 1.06, 0.94–1.20; p = 0.994), and cure (RR, 95% CI: 1.00, 0.97–1.02; p = 0.661) did not differ significantly between SDCT and BDCT. Substantial heterogeneity was observed between the trials regarding the pooled proportion of antibiotic use within the SDCT groups (I² = 97.7%; p < 0.001). This meta-analysis provides evidence that SDCT seems to be an adequate alternative to BDCT regarding udder health with a simultaneous reduction in antibiotic use. Limitations might arise because of the small number of studies included. Full article

The aim of this paper is to describe the association between selected dietary minerals and mastitis in dairy cows. Minerals are a group of nutrients with a proven effect on production and reproductive performance. They also strongly affect immune system function. In particular their deficiencies may result in immunosuppression, which is a predisposing factor for udder inflammation occurrence. The role of selected dietary minerals (including calcium, phosphorus, magnesium, selenium, copper and zinc) has been reviewed. Generally, minerals form structural parts of the body; as cofactors of various enzymes they are involved in nerve signaling, muscle contraction and proper keratosis. Their deficiencies lead to reduced activity of immune cells or malfunction of teat innate defense mechanisms, which in turn promote the development of mastitis. Special attention was also paid to minerals applied as nanoparticles, which in the future may turn out to be an effective tool against animal diseases, including mastitis. To conclude, minerals are an important group of nutrients, which should be taken into account on dairy farms when aiming to achieve high udder health status. Full article

Nanomaterials have been used for diagnosis and therapy in the human medical field, while their application in veterinary medicine and animal production is still relatively new. Nanotechnology, however, is a rapidly growing field, offering the possibility of manufacturing new materials at the nanoscale level, with the formidable potential to revolutionize the agri-food sector by offering novel treatment options for prevalent and expensive illnesses such as bovine mastitis. Since current treatments are becoming progressively more ineffective in resistant bacteria, the development of innovative products based on both nanotechnology and phytotherapy may directly address a major global problem, antimicrobial resistance, while providing a sustainable animal health solution that supports the production of safe and high-quality food products. This review summarizes the challenges encountered presently in the treatment of bovine mastitis, emphasizing the possibility of using new-generation nanomaterials (e.g., biological synthesized nanoparticles and graphene) and essential oils, as candidates for developing novel treatment options for bovine mastitis. Full article