Nanotechnology for effective bovine mastitis treatment: Applications in biosensors and vaccines

Simple SummaryOregano essential oil is a plant essential oil extracted from the plant oregano (Origanum vulgare), which has an antibacterial, anti-inflammatory, and antioxidant effect. Bovine mastitis is a process in which bacterial infection and inflammation co-exist. In this study, network pharmacology technology was used to explore the mechanism of oregano essential oil in the treatment of bovine mastitis, and to predict the core genes and key signaling pathways exerting therapeutic effects. The results showed that the key action targets of oregano essential oil in the treatment of dairy bovine mastitis were TNF, TLR4, ALB, IL-1β, TLR2, IL-6, IFNG, and MyD88, and the main signaling pathways were PI3K-Akt, MAPK, IL-17, NF-κ B. The molecular docking confirmed that thymol and carvacrol in oregano essential oil had a good binding ability to the core targets of TNF, IL-6, MyD88, and ALB. These results indicated that thymol and carvacrol had good anti-inflammatory effects. Therefore, oregano essential oil was considered to have a good therapeutic effect on bovine mastitis.The active components, potential targets, and mechanisms of action of oregano essential oil in the treatment of bovine mastitis disease were investigated using network pharmacology and molecular docking approaches. The TCMSP and literature databases were examined for the main compounds in oregano essential oil. Afterward, the physical, chemical, and bioavailability characteristics of the components were evaluated. The PubChem, BATMAN, PharmMapper, and Uniprot databases were utilized to predict the target genes of the major components of oregano essential oil. Via the databases of DrugBank, OMIM, GeneCards, TTD, and DisGenet, the disease targets of bovine mastitis were discovered. We analyzed common targets and built protein–protein interaction (PPI) networks using the STRING database. Key genes were analyzed, obtained, and compound–target–pathway–disease visualization networks were created using Cytoscape. For the GO function and KEGG pathway enrichment analysis, the DAVID database was utilized. Molecular docking via Autodock Tools was utilized to evaluate the reliability of the interactions between oregano essential oil and hub targets. Thymol, carvacrol, and p-cymene are the three major components found in oregano essential oil. The potential targets (TNF, TLR4, ALB, IL-1β, TLR2, IL-6, IFNG, and MyD88) were screened according to the visual network. The enrichment analysis suggested that the major signaling pathways in network pharmacology may include PI3K-Akt, MAPK, IL-17, and NF-κ B. Molecular docking analysis shows that thymol had good docking activity with TNF, IL-6, and MyD88, carvacrol had good docking activity with TNF, and p-cymene had good docking activity with ALB. This study clarified the mechanism of action of oregano essential oil in the treatment of bovine mastitis, thus providing data supporting the potential for the use of oregano essential oil in the development of new therapeutics for bovine mastitis.

The Development and Impact of Tuberculosis Vaccines

A novel formulation of cationic liposomes was studied by mixing dipalmitoylphosphatidylcholine (DPPC) with tetradecyltrimethylammonium bromide gemini surfactants with different alkane spacer groups lengths attached to their ammonium head-groups. The physicochemical characterization of the cationic liposomes was obtained by combining experimental results from differential scanning microcalorimetry (DSC) with molecular dynamic simulations, in order to understand their structural configuration. An adapted Ising model was used to interpret the results in terms of cooperativity of the phase transitions.The gemini surfactants partition into the lipid bilayer of DPPC liposomes, and the induced changes in colloidal stability and phase transition were analyzed in detail. The DPPC liposomes became positively charged upon gemini surfactant partition, showing increased colloidal stability. Our results show significant differences in structural configuration between gemini surfactants with short and long spacer lengths. While gemini with shorter spacers allocate within the lipid bilayer with both headgroups in the same layer, geminis with longer spacers unexpectedly intercalate in the lipid membrane in a particular zig-zag configuration, with each headgroup located at a different side of the bilayer, altering the coupling degree parameters of the membrane’s phase transition.The extraordinary increase of colloidal stability of DPPC liposomes with gemini surfactants at very low molar ratio and the possibility to tune the physicochemical properties of the membrane by control de spacer length of the geminis opens new possibilities for cationic liposomal formulations with potential applications in vaccines, drug/gene delivery or biosensing.

Bovine mastitis is the most important disease of dairy herds worldwide. Its main etiologic agents are bacteria, including Streptococcus agalactiae. The importance of this agent in bovine mastitis is because it is highly contagious and has a high impact on the occurrence of clinical mastitis cases and in the increase of the bulk milk somatic cell counts. The dry cow therapy and the treatment of the clinical mastitis cases stand out among the measures to control intramammary infections in cows. However, these strategies require knowledge about the antimicrobial susceptibility of the causal microorganisms. Thus, this study aimed to evaluate the antimicrobial susceptibility of 89 S. agalactiae strains isolated from bovine mastitis between the years 2004 and 2008 in dairy herds from Campo das Vertentes region, Minas Gerais State, Brazil. The disc diffusion technique was used and the antimicrobials currently used in mastitis therapy were tested. The isolates tested showed 100% susceptibility to chloramphenicol, ceftiofur, cefotaxime, enrofloxacin, and cefquinome. High frequencies of susceptibility (>95%) were also observed for the beta-lactams (penicillin G, ampicillin, and oxacillin), cephalosporins (cephalotin, ceftiofur, cefotaxime, cefoperazone, and cefquinome), florfenicol, gentamicin, lincomycin, nitrofurantoin, and sulfamethoprim. The strains showed high frequencies of resistance to neomycin (15.74%), and tetracycline (21.35%). Multidrug resistance was detected in 2.25% of the tested isolates. The results pointed to variations in the antimicrobial susceptibility profiles of the studied strains and the importance of the use of the susceptibility tests to determine the correct antimicrobial to be applied in the treatment of bovine mastitis caused by S. agalactiae. The high frequencies of resistance observed to some antimicrobials, such as neomycin and tetracycline, commonly used in the treatment of mastitis and other pathologies, highlighted the need for more judicious use of antimicrobials on dairy farms.

The topological conformation of DNA is a critical determinant of its biological function, but distinguishing between different isoforms, such as circular and linear, remains challenging. This study demonstrates the application of terahertz metamaterial spectroscopy as a powerful, label-free platform for the quantitative identification of DNA topology. Using bacteriophage ΦX174 single-stranded DNA as a model system, we show that a custom metamaterial sensor can generate distinct resonance frequency shifts for circular and linear topologies at concentrations as low as ng/μl. The spectral shifts exhibit a strong linear correlation with DNA concentration, confirming the quantitative capability, whereas, UV absorption spectroscopy fails to differentiate the isoforms. Our findings establish THz metamaterial spectroscopy as a robust tool for the direct analysis of DNA topology, with significant potential for applications in gene therapy, DNA vaccination, and biosensing.

Objective: to describe the importance of nanoparticles in the prevention, diagnosis and treatment of bovine mastitis. Design/ Methodology/ Approach: nanotechnology in the field of veterinary medicine has revolutionized scientific research. As it aims to access a new era in the diagnosis and treatment of disimile diseases such as bovine mastitis. In regard to it, this disease was characterized in its most common origin and diagnosis. Then, we examined state-of-the-art papers on the use of nanoparticles as an alternative treatment to verify results and effectiveness. Results: reviewed reports on the use of nanotechnology as treatment did show favorable, faster and more effective results. Also, it is discussed how nanotechnology has innovated the field of prevention of this disease that can occur in cattle in a clinical or subclinical form, in order to offer innovative solutions to current problems, such as the prevention, diagnosis, and treatment of this common disease in dairy cattle. Limitations/ implications of the study: despite its significant importance in the treatment of bovine mastitis. applied nanotechnology still has certain limitations to be noted. These include the lack of a single, standardized dose; doses used must be tested through clinical studies assessing some aspects such as nanoparticle type, drug load, and infection severity. In addition to other aspects like toxicity, distribution, bioavailability, immunological interactions, and costs. Findings/ Conclusions: the use of nanoparticles in veterinary medicine as treatment of bovine mastitis allows transforming delivery vehicles and assimilation paths for drugs and vaccines, which enhances treatment effectiveness. Therefore, the use of nanomaterials positively impacts animal health and well-being.

BackgroundMastitis is considered the most significant and persistent disease in dairy cows, bringing about large economic losses. Subclinical mastitis brings about major cost implications, for it is difficult to detect due to absence of any visible indications and can persist in the mammary tissue throughout lactation. Immunomodulators have been widely used to reduce intramammary infections by modulating bovine mammary gland. Atractylodis macrocephalae Koidz. polysaccharides (RAMP), extracted from herbal medicine, has been used widely especially for its immunomodulatory function for many years. The objective of this study was to estimate an oil emulsified Atractylodis macrocephalae Koidz. polysaccharides (RAMP-O) as a potential therapeutic agent to treat subclinical mastitis by subcutaneous injection of RAMP-O in the area of supramammary lymph node in lactating cows via analysis of SCC, IMIs and NAGase.ResultsInjection of RAMP-O in the area of supramammary lymph node significantly reduced milk SCC and NAGase activity compared with control. The quarters with bacterial infection were also progressively reduced in RAMP-O treated cows and only 9 quarters were found to have bacterial infection, while no obvious change was found in the control group.ConclusionsSubcutaneous injection of RAMP-O in the area of supramammary lymph node had therapeutic value in the treatment of bovine subclinical mastitis by reducing SCC, NAGase and IMIs in milk. Considering both the therapeutic effect and the cost of RAMP-O, 32 mg per dose was found most suitable to reduce milk SCC and NAGase. Therefore, RAMP-O deserves further study for its use in treatment of bovine mastitis.

Houttuynin sodium bisulphate (HSB), alpha hydroxyl-capryl-ethyl-sodium-sulphonate, is a product formed by reacting sodium bisulphate with houttuynin, which is obtained from a medicinal herb Houttuynia cordata Thunb. From HBS an aqueous intramammary solution was made for the treatment of bovine clinical mastitis. A total of 104 acute and subacute mastitis cases were randomly assigned into two groups with 52 cases in each group: 1. an HSB group in which 80 mg HSB was infused into an affected gland; and 2. a PS group in which intramammary administration of 800,000 i.u. penicillin G in combination with 1 g of streptomycin (PS) was conducted. The treatments were administered twice daily until the inflammatory signs were eliminated and mammary secretion became normal. In acute mastitis, 88.2% (15 of 17) were clinically cured and 52.9% (nine of 17) microbiologically cured by HSB; in the PS group, 90.0% (18 of 20) were clinically cured and 55.0% (11 of 20) microbiologically cured. In subacute cases, the clinical and microbiological cure rates were 94.3% (33 of 35) and 45.7% (16 of 35) respectively, in the HSB group; and in the PS group the clinical and microbiological cure rates were 93.7% (30 of 32) and 43.8% (14 of 32), respectively. No statistically significant difference was found between HSB and PS groups in the treatment of acute as well as subacute mastitis. In addition, an inhibitory effect was found on the growth of lactic streptococcus in the milk collected within 48 h of intramammary treatment with penicillin G in combination with streptomycin. However, for HSB, a mild inhibitory effect on lactic streptococci was detected in the milk within 12 h of treatment.

Mastitis occurs in numerous species. Antimicrobial agents are used for treatment of infectious mastitis in dairy cattle, other livestock, companion animals, and humans. Mastitis is an economically important disease of dairy cattle and most mastitis research has focused on epidemiology and control of bovine mastitis. Antibiotic treatment of clinical and subclinical mastitis in dairy cattle is an established component of mastitis control programs. Research on the treatment of clinical and subclinical mastitis in other dairy species such as sheep and goats has been less frequent, although the general principles of mastitis therapy in small ruminants are similar to those of dairy cattle. Research on treatment of clinical mastitis in humans is limited and as for other species empirical treatment of mastitis appears to be common. While antimicrobial susceptibility testing is recommended to direct treatment decisions in many clinical settings, the use of susceptibility testing for antibiotic selection for mastitis treatments of dairy cattle has been challenged in a number of publications. The principle objective of this review is to summarize the literature evaluating the question, "Does antimicrobial susceptibility predict treatment outcome for intramammary infections caused by common bacterial pathogens?" This review also addresses current issues related to antimicrobial use and treatment decisions for mastitis in dairy cattle. Information on treatment of mastitis in other species, including humans, is included although research appears to be limited. Issues related to study design, gaps in current knowledge and opportunities for future research are identified for bovine mastitis therapy.

Over the last few years, the author, alongside many other dairy vets, has been working with dairy farmers to support them in carrying out selective dry cow therapy in their herds, while ensuring that cow health and welfare are not compromised. We can now have more informed discussions about the selective treatment of clinical mastitis in lactating dairy cows. This has been enabled by the recent advances and availability of cow-side pathogen testing, with several publications in the scientific literature evaluating farms carrying out selective treatment of clinical mastitis compared to blanket treatment, as well as the improved on-farm data we have around mastitis cases and mastitis patterns in our herds. The advantage of selective treatment is predominantly the reduced antibiotic use aligned with the One Health approach to the responsible use of antibiotics. There are other advantages to the selective treatment of cases, including targeted (narrow spectrum) antibiotic use and the increased farm focus and close monitoring of mastitis cases and cure rates on farm that come with reviewing mastitis treatment protocols. Vets can also be actively engaged in discussing the treatment and monitoring outcomes of mild and moderate cases of clinical mastitis, an area that has often been overlooked as a result of the focus on sick cows or toxic/severe cases requiring more involved and tailored therapy. Rapid and accurate cow-side pathogen tests allow selective treatment of clinical mastitis to be carried out on farm without compromising on treatment delay, so that the most effective treatment can be given to the appropriate mastitis cases. The focus of mastitis control must always remain on preventative measures to reduce the number of new infections. Through on-farm engagement with a wider mastitis discussion and creating bespoke tailored mastitis treatment programmes as part of selective treatment of clinical mastitis, this can serve as a regular reminder that prevention of new infections will always lead to the most significant benefits. This article reviews the available information on the selective treatment of clinical mastitis and presents some initial steps in data gathering and evaluation to ensure that the appropriate farms are undertaking selective treatment of mastitis cases.

Over the last few years, the author, alongside many other dairy vets, has been working with dairy farmers to support them in carrying out selective dry cow therapy in their herds, while ensuring that cow health and welfare are not compromised. We can now have more informed discussions about the selective treatment of clinical mastitis in lactating dairy cows. This has been enabled by the recent advances and availability of cow-side pathogen testing, with several publications in the scientific literature evaluating farms carrying out selective treatment of clinical mastitis compared to blanket treatment, as well as the improved on-farm data we have around mastitis cases and mastitis patterns in our herds. The advantage of selective treatment is predominantly the reduced antibiotic use aligned with the One Health approach to the responsible use of antibiotics. There are other advantages to the selective treatment of cases, including targeted (narrow spectrum) antibiotic use and the increased farm focus and close monitoring of mastitis cases and cure rates on farm that come with reviewing mastitis treatment protocols. Vets can also be actively engaged in discussing the treatment and monitoring outcomes of mild and moderate cases of clinical mastitis, an area that has often been overlooked as a result of the focus on sick cows or toxic/severe cases requiring more involved and tailored therapy. Rapid and accurate cow-side pathogen tests allow selective treatment of clinical mastitis to be carried out on farm without compromising on treatment delay, so that the most effective treatment can be given to the appropriate mastitis cases. The focus of mastitis control must always remain on preventative measures to reduce the number of new infections. Through on-farm engagement with a wider mastitis discussion and creating bespoke tailored mastitis treatment programmes as part of selective treatment of clinical mastitis, this can serve as a regular reminder that prevention of new infections will always lead to the most significant benefits. This article reviews the available information on the selective treatment of clinical mastitis and presents some initial steps in data gathering and evaluation to ensure that the appropriate farms are undertaking selective treatment of mastitis cases.

Bovine mastitis is a common disease of dairy cows, which is the main reason for the economic loss of dairy industry. This paper reviews the types, etiology, diagnosis, prevention and treatment of bovine mastitis, in order to provide some reference basis for the prevention and treatment of bovine mastitis.

This chapter attempts to review the pathophysiology of bovine mastitis in the context of susceptibility, progression of disease, and the cytokine response of the host to various pathogens. It provides an overview of the presently available recombinant bovine cytokines and their corresponding in vitro and in vivo effects on the cellular activity of bovine phagocytic cells. A bridge between the pathophysiology of the disease process and the known structure-function relationships of the cytokines is then traversed by a discussion of the impact of exogenous administration of recombinant cytokines on the initiation and progression of infections using various bovine mastitis model systems. The chapter discusses the impact on the dairy industry with respect to the regulation and application of these potent bioresponse modifiers. A fundamental principle of designing the optimal approach for the prevention or treatment of bovine mastitis with immunomodulatory compounds is that it must be based on an understanding of the pathophysiology of the disease.

It has been a pleasure and honor to know Dr. Landheer for more than three decades and to have collaborated with him. In this invited presentation, I will focus on one aspect of our collaboration – the topic of Biosensors – which was our last research collaboration. In the list of references [1-15], I provide all publications we collaborated on since 1986.Biosensors are increasingly used in environmental applications, especially for water quality monitoring. This is because the availability of safe drinking water is fundamental to our good health. However, as water resources get increasingly stressed, ensuring a safe water supply and effective water treatment becomes increasingly important. In addition, waterborne illnesses are a significant public health problem. At the same time, current monitoring of microbiological contamination of water currently is time-consuming, laboratory based, and frequently compromises the timeliness of health advisory warnings even when contamination is found. Therefore, rapid detection of unsafe water can contribute greatly to mitigating the morbidity and mortality associated with waterborne diseases due to microbiological contaminants. Fortunately, the research community has shown increasing interest in the development of microtechnology-based sensors for the detection and identification of the bio-contaminants. These sensing systems use the same fabrication technology that has enabled the drastic lowering of cost, exponential increase in complexity of electronic chips and widespread availability of computing resources. In this presentation, we will discuss a low-cost, electrical, label-free microfabricated biosensor that we have been developing for pathogen detection related to water quality and also for ubiquitous-healthcare applications. The use of nano-dimensions devices to create futuristic nano-biosensors for both environmental and health applications will be introduced. And we will also describe our ongoing work to create highly integrated and parallel detection systems by integrating the sensor, the processing electronics and the pre-processing stages on the same cheap substrate. Finally, the success of such a low-cost, highly integrated sensing system demands a convergence of expertise from various engineering disciplines, the physical and life sciences as well as public health. References D Landheer et al, “Bioaffinity Sensors Based on MOS Field—Effect Transistors,” in Semiconductor Device-Based Sensors for Gas, Chemical, and Biomedical Applications, Eds. Ren, Pearton, Taylor & Francis Books, Boca Raton, 215-265, 2010.MW Shinwari, et al, Microfabricated Reference Electrodes and their Biosensing Applications, Sensors, Vol. 10(3), pp. 1679-1715, 2010.MW Shinwari, MJ Deen, D Landheer, “Study of the Electrolyte-Insulator-Semiconductor Field-Effect Transistor with Applications in Biosensor Design,” Microelectronics Reliability, Vol. 47(12), pp. 2025-2057, 2007.D Landheer, et al, Calculation of the Response of Field-Effect Transistors to Charged Biological Molecules, IEEE Sensors Journal, Vol. 7, 1233-1242, 2007.WH Jiang, et al, Post-processing of Commercial CMOS Chips for the Fabrication of DNA Bio-FET Sensor Arrays, Proceedings of MRS Symposium - Fall Meeting, 6 pages, 2006.Bioelectronics, Biointerfaces, and Biomedical Applications 2, Eds., D Landheer, R. Bashir, M. Deen, C. Kranz, C. Liu, M. Madou, A. Offenhaeusser, R. Schasfoort, ECS Transactions, Vol. 3, Issue 26, 2006.MJ Deen, et al, Noise Considerations in Field-Effect Biosensors, Journal Applied Physics, Vol. 100, #074703, 8 pages, 2006.MJ Deen, et al, High Sensitivity Detection of Biological Species via the Field-Effect, Proceedings of the IEEE ICCDCS, Playa del Carmen, Mexico, pp. 381-385, 2006.D Landheer, et al, Model for the Field-Effect from Layers of Biological Macromolecules on the Gates of Metal-Oxide-Semiconductor Transistors, Journal Applied Physics, Vol. 98, # 044701, 2005.Silicon Nitride and Silicon Dioxide Thin Insulating Films, Eds., R.E. Sah, MJ Deen, D Landheer, K.B. Sundaram, W.D. Brown, D. Misra, ECS Proceedings PV-03, 636 pages 2003.Silicon Nitride and Silicon Dioxide Thin Insulating Films, Eds., K.B. Sundaram, MJ Deen, D Landheer, W.D. Brown, D. Misra, M.D. Allendorf, R.E. Sah, ECS Proceedings Volume PV 2001-7, 2001.MJ Deen, et al, Low Frequency Noise in Cadmium Selenide Thin-Film Transistors, Applied Physics Letters, Vol. 77(14), pp. 2234-2236, 2000.MJ Deen, et al, Low Frequency Noise in CdSe Thin-Film Transistors, ESSDERC 2000, Cork, Ireland, pp. 592-595, 2000.MJ Deen, et al, NbN Thin Films Reactively Sputtered with a High Field DC Magnetron, Journal of Vacuum Science and Technology A, Vol. 6(4), pp. 2299-2303, 1988.MJ Deen, et al, The Effect of the Deposition Rate on the Properties of DC Magnetron Sputtered NbN Thin Films, Bull Am Phys Soc., Vol. 32(3), p. 646, 1987.

Mastitis is defined as inflammation of the mammary gland and is characterized by causing significant alterations in the composition of milk and economic losses and endless perjuries to the dairy industry. For this reason, and in view of the growing demand for antibiotic-free and quality products for human consumption, alternatives are being sought for current broad-spectrum antibiotic-based mastitis treatments. Within this framework, this article aims to demonstrate through a case report the effectiveness of the use of ozonized solutions in the treatment of clinical mastitis in dairy cows. For which a cow that presented clinical mastitis in 2 quarts was treated, with an ozonated solution (10 ml of solution in total, 5 ml of ozonated sunflower oil plus 5 ml of ozonated physiological serum in a concentration of 12 mg/ml), this treatment was compared with a usual commercial antibiotic treatment (Masticine L). At the end of the treatments, a favorable change was perceived in terms of visible physical aspects in both treatments, the antibiotic having a better performance in this regard, however the treatment with ozonized solution had favorable effects in terms of milk composition, improving its quality and lowering treatment costs therefore the use of intramammary ozonated solutions is recommended for the treatment of clinical mastitis in dairy cows.