Mastitis In Cows Research Articles

Succinate exacerbates mastitis in mice via extracellular vesicles derived from the gut microbiota: a potential new mechanism for mastitis.

A high grain diet causes an ecological imbalance in the gut microbiota and serves as an important endogenous trigger of mastitis in dairy cows, but the underlying mechanisms are unclear. Our previous study revealed that subacute rumen acidosis (SARA)-associated mastitis has distinct metabolic profiles in the rumen, especially a significant increase in succinate, but the role of succinate in the pathogenesis of mastitis remains unclear. Succinate treatment exacerbates low-grade endotoxemia-induced mastitis in mice. Specifically, succinate increased the production of gut microbiota-extracellular vehicles (mEVs) containing lipopolysaccharides, which can diffuse across the damaged intestinal barrier into the mammary glands. Administration of mEVs promotes mammary inflammation via activation of the TLR4/NF-κB pathway. Our findings suggest that succinate promotes mastitis through the proliferation of enteric pathogens and mEVs production, suggesting a potential strategy for mastitis intervention on the basis of intestinal metabolic regulation and pathogen inhibition. The role of mEVs in interspecific communication has also been elucidated.

MastitisApp: a software for preventive diagnosis of mastitis in dairy cows

Dairy farming has great economic value in Brazil, however, during production, diseases such as mastitis can occur in animals, which can reduce productivity and, consequently, economic profitability. When mastitis is present in animals, it can cause physical and chemical changes in the milk, affecting its quality, market value and also compromising the health of the animal. MastiteApp is a tool to help producers prevent mastitis in their herds by checking the temperature taken from the four teats of the animal. To perform the analysis, the temperature of all the animals' teats must be measured and, if there is a change in temperature, the system will display a message informing the producer of the possible presence of subclinical mastitis in their animal. The application has proven to be efficient in alerting producers to the possible presence of subclinical mastitis in the first few days of manifestation, thus initiating treatment and preventing the disease from worsening.

Blood-feeding stable flies, Stomoxys calcitrans (Diptera: Muscidae), are attracted to, and transmit Staphylococcus aureus (Bacillales: Staphylococcaceae), a causal agent of bovine mastitis: a laboratory pilot study.

Stable flies, Stomoxys calcitrans (Linnaeus) (Diptera: Muscidae), are common blood-feeding ectoparasites of cows and thus potential vectors of the skin-dwelling bacterium Staphylococcus aureus, a causal agent of bovine mastitis which inflicts udder inflammation in cows. Our objectives were to determine whether stable flies (i) are attracted to disease-causing strains of S. aureus, and (ii) transmit S. aureus from infected blood to sterile blood. In 3-chamber olfactometers, five of eight S. aureus strains grown on agar and tested versus sterile agar attracted female stable flies. When flies ingested droplets of blood inoculated with S. aureus at doses of 0 (control), 105 (low), 107 (medium), and > 109 (high) colony-forming units per milliliter and subsequently ingested sterile blood, they transmitted S. aureus to the sterile blood. The dose of S. aureus in blood droplets fed upon by flies during their first feeding bout dose-dependently affected the amount of bacteria that flies transmitted to sterile blood during their second feeding bout, but the time elapsed between feeding bouts (0h, 1h, 8h, and 24h) had no effect on the amount of microbes transmitted to sterile blood. Our data infer the existence of a positive feedback loop. First, stable flies carrying S. aureus and feeding on cows transmit S. aureus, thereby causing mastitis. As S. aureus bacteria of afflicted cows proliferate, they attract even more flies which, in turn, worsen the infection. This type of feedback loop underscores the need for effective stable fly control tactics that curtail the incidence of bovine mastitis in cows.

Milk metabolite profiling of dairy cows as influenced by mastitis

Mastitis is a disease with frequent incidence in dairy cows, causing huge financial losses to the dairy industry globally. The identification of certain biomarkers is crucial for the early diagnosis and management of mastitis. Metabolomics technology is a useful tool to accurately and efficiently analyze the changes of metabolites in biofluids in response to internal and external stimulations. Milk is the secreted by udder, and milk metabolites can directly reflect whether the udder are in the healthy or diseased state. The milk metabolomics analysis of mastitis can reveal the physiological and pathological changes of mammary gland and screen the related biomarkers, so as to offer useful reference for the prediction, diagnosis, and management of mastitis. Therefore, the aim of the present study was to comprehensively summarize milk metabolic change caused by naturally occurring or experimentally induced mastitis in dairy cows. In addition, comparative analysis and enrichment analysis were conducted to further discover potential biomarkers of mastitis and to identify the relevant pathways differentiating the healthy and mastitic cows. Multiple milk metabolites were identified to be altered during mastitis based on different metabolomics platforms. It was noteworthy that there were 28 metabolites not only identified by at least two different studies, but also showed consistent change tendency among the different studies. By comparison with literature, we further identified 12 milk metabolites, including acetate, arginine, β-hydroxybutyrate, carnitine, citrate, isoleucine, lactate, leucine, phenylalanine, proline, riboflavin, and valine that were linked with the occurrence of mastitis, which suggested that these 12 milk metabolites could be potential biomarkers of mastitis in dairy cows. Several pathways were revealed to explain the mechanisms of the variation of milk metabolites caused by mastitis, such as phenylalanine, tyrosine and tryptophan biosynthesis, arginine and proline metabolism, riboflavin metabolism, and tricarboxylic acid (TCA) cycle. These results offer a further understanding for the alteration of milk metabolites caused by mastitis, which have a potential significance in the development of more reliable biomarkers for mastitic diagnosis in dairy cows.

Mycoplasma bovis 5'-nucleotidase is a virulence factor conferring mammary fitness in bovine mastitis.

Nucleases and 5' nucleotidase (5'-NT) play essential roles in cell biology and are often associated with bacterial virulence. In Mycoplasma spp., which have limited metabolic capacities and rely on nutrient availability, these enzymes are of significant importance for nucleotide salvage. This study explores the potential role of 2 membrane-associated lipoproteins, the major nuclease MnuA and 5'-NT, in Mycoplasma bovis mastitis. Mutants deficient in MnuA (mnuA::Tn) and in 5'-NT (0690::Tn) were identified through genome-wide transposon mutagenesis of M. bovis PG45 type strain and their fitness and virulence were assessed both in vitro, in axenic medium, and in vivo, using murine and cow mastitis models. The mnuA::Tn mutant demonstrated reduced nuclease activity, while 0690::Tn exhibited slow log-phase growth and impaired hydrolase activity towards nucleotides as well as deoxynucleotides (dAMP and dGMP). In comparison to the parent strain, the 0690::Tn mutant displayed markedly reduced fitness, as evidenced by a significant decrease or even absence in post-challenge mycoplasma counts in murine and cow mammary tissues, respectively. Moreover, the 0690::Tn mutant failed to induce mastitis in both experimental models. Conversely, the mnuA::Tn mutant induced inflammation in murine mammary glands, characterized by neutrophil infiltration and increased expression of major inflammatory genes. In cows, the mnuA::Tn was able to cause an increase in somatic cell counts in a manner comparable to the wild type, recruit neutrophils, and induce mastitis. Collectively, these findings provide complementary insights, revealing that disruption of 5'-NT significantly attenuated M. bovis pathogenicity, whereas a MnuA-deficient mutant retained the ability to cause mastitis.

Synergistic Antibacterial Effect of Lactic Acid Bacteria and Baicalin Against Staphylococcus aureus In Vitro and In Vivo.

Pathogenic bacteria such as Staphylococcus aureus (S. aureus) are the principal cause of cow mastitis, which primarily impacts milk yield and results in significant financial losses for the animal husbandry industry. Lactic acid bacteria-cell-free supernatant (LAB-CFS) and baicalin (BAI) both have a number of biological effects, including decreasing inflammation. The combined use of LAB-CFS and BAI does not appear to have been used toprotective against mastitis, however, and the underlying mechanisms are yet unknown. In this study, in vitro activity of LAB-CFS and BAI alone and in combination was determined (checkerboard experiments, time-kill curves, and flow cytometry to investigate membrane permeability) and examined the protective effects of LAB-CFS and BAI on S. aureus-induced mastitis in mice and the impact of NF-κB signaling pathways on the emergence of mastitis. We discovered that when LAB-CFS and BAI were used together, S. aureus was more effectively treated than when LAB-CFS and BAI were used separately. Flow cytometry demonstrated that LAB-CFS and BAI work together to kill bacteria. In vivo, the usage of LAB-CFS and BAI decreased the activity of myeloperoxidase, as well as IL-6, IL-1β, and TNF-α secretion and the levels of TLR2 and p65 (NF-κB) expression. These findings suggested that LAB-CFS and BAI had a preventive effect against mastitis brought on by S. aureus. Therefore, the NF-κB signaling pathway is thought to be the likely mechanism through which LAB-CFS and BAI reduced S. aureus-induced inflammation in the mammary of cows. For the treatment of cow mastitis, LAB-CFS and BAI are likely to replace antibiotics.

A New Way to Identify Mastitis in Cows Using Artificial Intelligence

Mastitis is a disease that is considered an obstacle in dairy farming. Some methods of diagnosing mastitis have been used effectively over the years, but with an associated relative cost that reduces the producer’s profit. In this context, this sector needs tools that offer an early, safe, and non-invasive diagnosis and that direct the producer to apply resources to confirm the clinical picture, minimizing the cost of monitoring the herd. The objective of this study was to develop a predictive methodology based on sequential knowledge transfer for the automatic detection of bovine subclinical mastitis using computer vision. The image bank used in this research consisted of 165 images, each with a resolution of 360 × 360 pixels, sourced from a database of 55 animals diagnosed with subclinical mastitis, all of which were not exhibiting clinical symptoms at the time of imaging. The images utilized in the sequential learning transfer were those of MammoTherm, which is used for the detection of breast cancer in women. The optimized model demonstrated the most optimal network performance, achieving 92.1% accuracy, in comparison to the model with manual search (86.1%). The proposed predictive methodologies, based on knowledge transfer, were effective in accurately classifying the images. This significantly enhanced the automatic detection of both healthy animals and those diagnosed with subclinical mastitis using thermal images of the udders of dairy cows.

Transcriptome analysis identifies the NR4A subfamily involved in the alleviating effect of folic acid on mastitis induced by high concentration of Staphylococcus aureus lipoteichoic acid

BackgroundStaphylococcus aureus (S. aureus) mastitis results in economic losses during dairy production. Understanding the biological progression of bovine S. aureus mastitis is vital for its prevention. Lipoteichoic acid is a key virulence factor of S. aureus (aLTA), but the main biological pathways involved in its effect on bovine mammary epithetionallial cells (Mac-T) apoptosis and necrosis have not been fully explored. Folic acid (FA) has anti-inflammatory and anti-apoptotic effects. However, the role of FA in mediating the effects of aLTA on apoptosis and necrosis remains unknown.ResultsWe found that low concentration of aLTA inhibited apoptosis and necrosis and that high concentration promoted the apoptosis and necrosis of Mac-T. FA pretreatment alleviated high concentration of aLTA induced apoptosis. Through transcriptomic analysis, we found that nuclear receptor subfamily 4 group A (NR4A), which alters the expression of downstream genes involved in apoptosis, proliferation, and inflammation, decreased under stimulation with a low concentration of aLTA and increased under stimulation with a high concentration of aLTA. Under stimulation with a high concentration of aLTA, the expression of the NR4A subfamily could be inhibited by FA. The results showed that aLTA may affect apoptosis and necrosis through the NR4A subfamily by targeting genes involved in bacterial invasion of epithelial cells, the IL-17 signaling pathway, DNA replication, longevity regulation, the cell cycle, and tight junction pathways. We further found that the expression trends of NR4A1 and the target genes of the NR4A subfamily (PTGS2, ESPL1, MCM5, and BUB1B) in the blood of healthy cows (Healthy), subclinical mastitis cows (SCM), and SCM supplemented with FA (SCM_FA) were consistent with those observed at the cellular level in this study.ConclusionsOur study revealed that low and high concentrations of aLTA have opposite effects on apoptosis and necrosis of Mac-T and that FA can alleviate the apoptosis induced by high concentration of aLTA. Transcriptome analysis revealed that the NR4A subfamily play a role in the ability of FA to alleviate the apoptosis and necrosis induced by high concentration of aLTA.

Diversity of Staphylococcus aureus enterotoxin genes and its potential impact on severity of mastitis in dairy cows.

Staphylococcus aureus enterotoxins (SEs) can lead to food poisoning and appear to play a pivotal role in the development and severity of mastitis. This study aimed to evaluate whether the presence and diversity of S. aureus Enterotoxin genes mastitis-causing are associated with the development and severity of mastitis in dairy cows. Comparative analysis of S. aureus isolates from cows with clinical and subclinical mastitis (103 isolates each, CM and SM respectively) was conducted, examining the occurrence of enterotoxins (sea-see, seg-sez, sel26, sel27, sel01 and se02). Clinical isolates exhibited greater diversity, with 14 genes compared to four in subclinical isolates, and higher gene frequency (100% vs. 86.4%). Among the classical SEs, only sec (1%) and sed (4.9%) were detected in clinical isolates, suggesting a potential insignificance in the development of mastitis. Conversely, the selw and selx genes were among the most prevalent in both clinical and subclinical mastitis isolates. While their role in human food poisoning has not been definitively established, they appear to influence mastitis development. Statistical analysis revealed significant differences in clinical isolates carrying seg, seh, sei, seo, seul1, seul2, selw, and selx genes compared to subclinical isolates, suggesting their potential involvement in the severity of mastitis. Determining the frequency of SE gene and its role in mastitis establishment and in clinical mastitis severity, as well as understanding how enterotoxins contribute to pathogenicity in clinical and subclinical mastitis, is crucial for guiding optimal medical therapy for animals and establishing effective veterinary treatments to mitigate economic losses in the dairy industry.

AMP-activated protein kinase mediates (-)-epigallocatechin-3-gallate (EGCG) to promote lipid synthesis in mastitis cows

Mastitis, a serious threat to the health and milk production function of dairy cows decreases milk quality. Blood from three healthy cows and three mastitis cows were collected in this study and their transcriptome was sequenced using the Illumina HiSeq platform. Differentially expressed genes (DEGs) were screened according to the |log2FoldChange| > 1 and P-value < 0.05 criteria. Pathway enrichment and functional annotation were performed through KEGG and GO analyses. Finally, the mechanism of the AMP-activated protein kinase (AMPK) mediation of (-)-epigallocatechin-3-gallate (EGCG) to promote lipid metabolism in mastitis cows was analyzed in bovine mammary epithelial cells (BMECs). Transcriptome analysis revealed a total of 825 DEGs, with 474 genes showing increased expression and 351 genes showing decreased expression. The KEGG analysis of DEGs revealed that they were mainly linked to tumour necrosis factor, nuclear factor-κB signalling pathway, and lipid metabolism-related signalling pathway, whereas GO functional annotation found that DEGs were enriched in threonine and methionine kinase activity, cellular metabolic processes, and cytoplasm. AMPK expression, which is involved in several lipid metabolism pathways, was downregulated in mastitis cows. The results of in vitro experiments showed that the inhibition of AMPK promoted the expression of lipid synthesis genes in lipopolysaccharide-induced BMECs and that EGCG could promote lipid synthesis by decreasing the expression of AMPK and downregulating the expression of inflammatory factors in inflammatory BMECs. In conclusion, our study demonstrated that AMPK mediated EGCG to inhabit of inflammatory responses and promote of lipid synthesis in inflammatory BMECs.

Microbial dysbiosis in the gut–mammary axis as a mechanism for mastitis in dairy cows

Mastitis is a significant and costly disease in dairy cows, reducing milk production and affecting herd health. Recent research highlights the role of gastrointestinal microbial dysbiosis in the development of mastitis. This review focuses on how microbial imbalances in the rumen and intestines can compromise the integrity of the gastrointestinal barriers, allowing harmful bacteria and endotoxins, such as lipopolysaccharide, to enter the bloodstream and reach the mammary gland, triggering inflammation. This process links gastrointestinal health to mammary gland inflammation through the gut–mammary axis. Furthermore, disruptions in glucose metabolism and immune responses are implicated in the progression of mastitis. This review underscores the potential for non‐antibiotic interventions aimed at restoring microbial balance to reduce mastitis incidence, providing new insights into improving dairy cow health and farm productivity. Our findings emphasise the critical need to explore preventive measures targeting the rumen and intestinal microbiota for effective mastitis control.

Genetic framework sequencing analysis of Candida tropicalis in dairy cow mastitis and study of pathogenicity and drug resistance

Candida tropicalis (C. tropicalis) is a zoonotic pathogen that is widespread in the environment and in recent years an increasing number of dairy cows have been infected with the fungus causing mastitis in cows.In this study, 37 milk samples from the udders of cows with clinical mastitis were collected from a dairy farm in Guangxi Province, China, from which C. tropicalis was isolated and identified, and then the isolated fungi were subjected to genome frame map sequencing, genome functional analysis as well as comparative genome analysis of the sequencing results, and combined with the virulence test of the fungi and drug sensitivity test of the fungi determined in infected mice, the resistance genes and pathogenicity of the fungi were Analysis of resistance genes and pathogenicity.Our study results revealed the isolation and characterisation of C. tropicalis from diseased cows, with a genome length of approximately 14.27 Mb. Functional annotation of the genome identified 4068 genes associated with C. tropicalis. The strain exhibited a chemoresistance mutation in the gene cyp51,a virulence-enhancing mutation in the gene VTC4, and mutations in genes linked to drug resistance. Pathogenicity tests demonstrated that C. tropicalis could induce damage to the internal organs of mice, leading to different levels of cyanosis in the abdominal cavity, white necrotic foci on the surface of internal organs, lung hemorrhage, and enlargement of the spleen and thymus.Histological sections also revealed varying degrees of hemorrhage and degenerative changes in the cells of different organs in the mice. Drug sensitivity tests showed that the fungus was highly sensitive to nystatin and ketoconazole, moderately sensitive to amphotericin B, and insensitive to antibiotics such as itraconazole, gentamicin, and penicillin. In conclusion, C. tropicalis isolated from dairy cows in the Guangxi region in this study was pathogenic and resistant to azoles such as itraconazole and fluconazole, and this study provides a theoretical basis for the further screening of novel resistance genes in C. tropicalis, as well as providing a certain reference for the drugs used for the treatment of fungal cow mastitis in this region.

Whole genome sequence analysis of multi-drug resistant and biofilm-forming Staphylococcus haemolyticus isolated from bovine milk

BackgroundMilk is an excellent growth medium for microorganisms due to its nutritive composition. Microorganisms have been implicated in bovine mastitis (BM) in dairy cows as well as causing infections in animals and humans. Despite extensive endeavours to manage BM, this condition continues to persist as the most prevalent and economically burdensome problem affecting dairy cattle on a global scale. Non-aureus staphylococci (NAS) species such as Staphylococcus haemolyticus, S. epidermidis, and S. xylosus are currently the predominant microbiological agents identified as the main cause of subclinical udder infections and are also considered opportunistic pathogens in cases of clinical mastitis in dairy cows. Therefore, it is crucial to elucidate the genetic profile of these species. The primary objective of this study was to characterise three phenotypically determined multidrug-resistant NAS environmental strains (NWU MKU1, NWU MKU2, and NWU MKS3) obtained from dairy cows milk via whole-genome sequencing.ResultsThe results confirmed that the three isolates were S. haemolyticus with genome sizes of 2.44, 2.56, and 2.56 Mb and a G + C content of 32.8%. The genomes contained an array of antibiotic resistance genes that may potentially confer resistance to a range of antibiotic classes, such as macrolides, fluoroquinolones, aminoglycosides, cephalosporins, tetracyclines, peptides, and phenicol. Furthermore, all the genomes carried virulence genes, which are responsible for several functions, such as adhesion, enzyme and toxin production. The genomes of these organisms contained signatures encoding mobile genetic elements such as prophages and insertion sequences.ConclusionThese findings indicate there is a need for diligent monitoring with improved management practices and quality control strategies on farms to safeguard milk production systems and human health.

Effect of Subclinical Mastitis Detected in the First Month of Lactation on the Reproductive Performance of Dairy Cows in Western Algeria

Background: The current study aimed to detect in the first month of lactation the prevalence and risk factors of subclinical mastitis (SCM) and its effects on reproductive performance in dairy cows in western Algeria. Methods: 530 cows from 18 herds were examined in the study. California Mastitis Test (CMT) was used to screen subclinical mastitis for individual cows and a score of two or more for any quarter without any clinical symptoms and abnormalities in milk was considered as positive for SCM. Milk samples positive for subclinical mastitis were subjected to bacteriological analysis. Result: The present study revealed a prevalence of subclinical mastitis of 34.90%, (185 of 530). The most common pathogen identified from CMT-positive milk samples was Staphylococcus aureus with 35.62%. Logistic regression analysis showed that parity, farming system and milking method affect significantly the prevalence of SCM. Subclinical mastitis during the first month of lactation affected significantly the reproductive performance by increasing calving to first service interval (CFSI), calving-to-conception interval (CCI)and number of services per conception (NSPC). The results of this study highlight the importance of mastitis control programs in dairy farms by showing that a high frequency of subclinical mastitis caused by Staphylococcus aureus may have a negative impact on the reproductive performance of dairy cows.

Prevalence and Bacterial Isolation Causing Clinical and Subclinical Mastitis in Egyptian Dairy Cow in Kafer El-Sheikh, Egypt

Prevalence and Bacterial Isolation Causing Clinical and Subclinical Mastitis in Egyptian Dairy Cow in Kafer El-Sheikh, Egypt

TMT-Based Quantitative Proteomics Revealed the Antibacterial Mechanism of Cinnamaldehyde against MRSA.

Natural plant extracts have demonstrated significant potential in alternative antibiotic therapies. Cinnamaldehyde (CA) has garnered considerable attention as a natural antibacterial agent. In this study, Tandem mass tag (TMT) quantitative proteomics combined with Western blot and RT-qPCR methods were employed to explore the antibacterial mechanism of CA against Methicillin-Resistant Staphylococcus aureus (MRSA) at the protein level. The results showed that a total of 254 differentially expressed proteins (DEPs) were identified in the control group and CA treatment group, of which 161 were significantly upregulated and 93 were significantly downregulated. DEPs related to nucleotide synthesis, homeostasis of the internal environment, and protein biosynthesis were significantly upregulated, while DEPs involved in the cell wall, cell membrane, and virulence factors were significantly downregulated. The results of GO and KEGG enrichment analyses demonstrated that CA could exert its antibacterial effects by influencing pyruvate metabolism, the tricarboxylic acid (TCA) cycle, teichoic acid biosynthesis, and the Staphylococcus aureus (S. aureus) infection pathway in MRSA. CA significantly inhibited the expression of recombinant protein MgrA (p < 0.05), significantly reduced the mRNA transcription levels of mgrA, hla, and sdrD genes (p < 0.05), and thermostability migration assays demonstrated that CA can directly interact with MgrA protein, thereby inhibiting its activity. These findings suggest that CA exerts its antibacterial mechanism by regulating the expression of related proteins, providing a theoretical basis for further development of clinical applications of antimicrobial agents derived from natural plant essential oils in the treatment of dairy cow mastitis.

Impact of a teat disinfectant based on Lactococcus cremoris on the cow milk proteome

BackgroundDairy cow milking practices require cleaning and disinfection of the teat skin before and after milking to ensure the safety and quality of milk and prevent intramammary infections. Antimicrobial proteins of natural origin can be valuable alternatives to traditional disinfectants. In a recent field trial, we demonstrated that a teat dip based on a nisin A-producing Lactococcus cremoris (L) had comparable efficacy to conventional iodophor dip (C) in preventing dairy cow mastitis. Here, we present the differential shotgun proteomics investigation of the milk collected during the trial.MethodsFour groups of quarter milk samples with low (LSCC) and high somatic cell count (HSCC) collected at the beginning (T0) and end (TF) of the trial were analyzed for a total of 28 LSCC (14 LSCC T0 and 14 LSCC TF) and 12 HSCC (6 HSCC T0 and 6 HSCC TF) samples. Milk proteins were digested into peptides, separated by nanoHPLC, and analyzed by tandem mass spectrometry (LC-MS/MS) on an Orbitrap Fusion Tribrid mass spectrometer. The proteins were identified with MaxQuant and interaction networks of the differential proteins were investigated with STRING. The proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD045030.ResultsIn healthy milk (LSCC), we detected 90 and 80 differential proteins at T0 and TF, respectively. At TF, the Lactococcus group showed higher levels of antimicrobial proteins. In mastitis milk (HSCC), we detected 88 and 106 differential proteins at T0 and TF, respectively. In the Lactococcus group, 14 proteins with antimicrobial and immune defense functions were enriched at TF vs. 4 proteins at T0. Cathelicidins were among the most relevant enriched proteins. Western immunoblotting validation confirmed the differential abundance.ConclusionsAt T0, the proteomic differences observed in healthy milk of the two groups were most likely dependent on physiological variation. On the other hand, antimicrobial and immune defense functions were higher in the milk of cows with mammary gland inflammation of the Lactococcus-treated group. Among other factors, the immunostimulatory action of nisin A might be considered as a contributor.

In vitro capsule or biofilm formation of Streptococcus uberis and bacteriological cure of bovine mastitis.

The relationship between the in vitro detected virulence factors biofilm and capsule formation of Streptococcus (S.) uberis isolates of clinical mastitis in dairy cows and the bacteriological cure rate after antibiotic therapy was investigated in order to better understand the importance of these virulence factors for the bacteriological cure rate. A total of 111 clinical mastitis (CM) cases were collected, in which S. uberis was bacteriologically detected. All mastitis cases were treated in accordance with the approval conditions of the antibiotic udder tubes used. Individual cow information including age, number of lactations, current lactation mastitis and antimicrobial treatment received was recorded. The microtiter plate test was used to detect biofilm formation and Anthony capsule staining was used to detect capsular capacity. Statistical analyses were performed to characterize the correlation between in vitro virulence factors and bacteriological cure (BC) rate. 30.5% (n=29) of the S. uberis isolates of bacteriologically cured cases and 34.5% (n=10) of the isolates of bacteriologically non-cured mastitis cases were found to have the ability to produce capsules in vitro. 70.7% (n=58) of the S. uberis isolates from bacteriologically cured mastitis cases had the ability to produce biofilm in vitro, whereas 58.6% (n=17) of the isolates of non-cured mastitis cases showed ability in producing biofilm. No correlation was found between the in vitro ability of S. uberis to form capsules and biofilms and the BC rate after antibiotic treatment of bovine mastitis. The present work has shown that the investigated in vitro virulence factors are not associated with the BC after antibiotic therapy. Further studies on the role of S. uberis virulence factors are needed to complete the missing knowledge on the difficulties in curing S. uberis mastitis. This study is of great clinical relevance since it enhances the understanding of the occurrence of BC in S. uberis mastitis cases. The investigated virulence factors are often addressed as possible reasons for therapy failure, although respective scientific studies are missing.

Proposal for the design of studies of intramammary antimicrobials to be conducted in Brazil for registration with the Ministry of Agriculture and Livestock

Before being made available to the Brazilian market, all veterinary medicines must be registered with the Ministry of Agriculture and Livestock after their efficacy, safety, and quality have been proven through clinical studies and scientific literature. Depending on the product class, the studies required by the regulatory authority must be carried out per Brazilian regulations and international reference guides. Some international organizations pursue the harmonization of these requirements, aiming at the mutual acceptance of studies conducted in different regions to facilitate international trade and reduce the use of animals in clinical research. Mastitis is one of the most prevalent and costly diseases in dairy cows, and it is associated with negative impacts on milk production and quality, cow welfare, and the profitability of the dairy industry. Normative Instruction No. 26/2009 determines a few rules for conducting studies for registering intramammary antimicrobials for cows with mastitis. However, the regulated sector and researchers report difficulties in following the recommendations of specific guidelines, which complement this regulation due to the peculiarities of Brazil’s production systems. This review article aims to provide subsidies and orientations that scientifically support the conduction and critical analysis of clinical studies proving the efficacy of intramammary products for treating clinical and subclinical mastitis in cows. Considering the need for scientific rigor in the studies, the recommendations available in international guidelines, and the need to adapt the protocols to the current situation of veterinary clinical research in Brazil, this document is intended to contribute to the internal harmonization of experimental protocols that support both the regulated sector and the regulatory authority.

Effect of Intramammary Hypochlorous Acid Administration on Subclinical Mastitis in Dairy Cows

Effect of Intramammary Hypochlorous Acid Administration on Subclinical Mastitis in Dairy Cows