Efficacy Assessment of Phage Therapy in Treating Staphylococcus aureus-Induced Mastitis in Mice.

The primary aim of this study was to evaluate the efficacy of phage against mastitis induced by drug-resistant S. aureus in a mouse model. In this study, five S. aureus phages-4086-1, 4086-2, 4086-3, 4086-4, and 4086-6-were isolated from milk samples secreted by mastitis cows. Transmission electron microscopy showed that all the five phages had icosahedral heads and short non-contractile tails, which are typical characteristics of the family Podoviridae. All these phages were species-specific against S. aureus. The one-step growth curve showed a short latency period (10-20 min) and high burst size (up to 400 PFU/infected cell). To evaluate the effectiveness of the phage 4086-1 in the treatment against mastitis, a mouse model of mastitis was challenged with drug-resistant S. aureus. The results showed the proliferation of S. aureus in the mammary glands was significantly inhibited after treating by phage 4086-1. The concentrations of TNF-α and IL-6 decreased significantly, which demonstrated the phages could effectively alleviate the inflammatory responses. Furthermore, the histopathological analysis showed that inflammatory infiltration in the mammary glands was significantly reduced. These results demonstrate that phage may be a promising alternative therapy against mastitis caused by drug-resistant S. aureus.

Advances in therapeutic and managemental approaches of bovine mastitis: a comprehensive review.

Mastitis (intramammary inflammation) caused by infectious pathogens is still considered a devastating condition of dairy animals affecting animal welfare as well as economically incurring huge losses to the dairy industry by means of decreased production performance and increased culling rates. Bovine mastitis is the inflammation of the mammary glands/udder of bovines, caused by bacterial pathogens, in most cases. Routine diagnosis is based on clinical and subclinical forms of the disease. This underlines the significance of early and rapid identification/detection of etiological agents at the farm level, for which several diagnostic techniques have been developed. Therapeutic regimens such as antibiotics, immunotherapy, bacteriocins, bacteriophages, antimicrobial peptides, probiotics, stem cell therapy, native secretory factors, nutritional, dry cow and lactation therapy, genetic selection, herbs, and nanoparticle technology-based therapy have been evaluated for their efficacy in the treatment of mastitis. Even though several strategies have been developed over the years for the purpose of managing both clinical and subclinical forms of mastitis, all of them lacked the efficacy to eliminate the associated etiological agent when used as a monotherapy. Further, research has to be directed towards the development of new therapeutic agents/techniques that can both replace conventional techniques and also solve the problem of emerging antibiotic resistance. The objective of the present review is to describe the etiological agents, pathogenesis, and diagnosis in brief along with an extensive discussion on the advances in the treatment and management of mastitis, which would help safeguard the health of dairy animals.

In vitro and in vivo assessment of phage therapy against Staphylococcus aureus causing bovine mastitis.

OBJECTIVE: The aim of this study was to assess the efficacy of lytic bacteriophages on Staphylococcus aureus causing bovine mastitis, by in vitro and in vivo assays using Galleria mellonella and murine mastitis models. METHODS: Between May and December 2016, ten S. aureus (five methicillin-resistant and five methicillin-sensitive) isolates were isolated from milk samples of cattle with mastitis in Belgium and Norway. The isolates were assessed in vitro for their susceptibility to four lytic bacteriophages (Romulus, Remus, ISP and DSM105264) and subsequently in vivo in G. mellonella larvae and in murine mastitis model. RESULTS: Romulus, Remus and ISP showed a lytic activity against the S. aureus isolates in vitro. A larvae survival rate below 50% was observed at 4 days post-inoculation (DPI) in the groups infected with a methicillin-sensitive S. aureus isolate and treated with these three phages in vivo. An incomplete recovery of the mouse mastitis was observed at 48h post-inoculation (HPI) in the groups infected and treated with the ISP phage in vivo. CONCLUSIONS: The observations are much more pronounced statistically between the infected- phosphate buffered saline (PBS)-treated and infected-phage-treated groups in G. mellonella and the murine mastitis model demonstrating an effect of the phages against S. aureus associated with bovine mastitis.

Assessment of acoustic pulse therapy (APT), a non-antibiotic treatment for dairy cows with clinical and subclinical mastitis.

Clinical and subclinical mastitis affects 30% of cows and is regarded as the most significant economic burden on the dairy farm reducing milk yield and quality and increasing culling rate. A proprietary Acoustic Pulse Therapy (APT) device was developed specifically for treating dairy cows. The APT device was designed to produce deep penetrating acoustic pulses that are distributed over a large treated area at a therapeutic level. This paper presents findings from a clinical assessment of this technology for the treatment of dairy cows with subclinical and clinical mastitis. In subclinical mastitis, a group of 116 cows from 3 herds were identified with subclinical intramammary infection and enrolled in the study; 78 cows were assigned to the treatment group and 38 cows to the control group. Significant differences (P<0.001) were found where 70.5% of the cows in the treatment group returned to normal milk production, compared with only 18.4% of the control group. Daily milk yields of the treated cows increased significantly (P<0.05) and the percentage of cows with log somatic cell count under 5.6 cells/mL was significantly higher (P<0.001). Milk of the infected quarters appeared normal with lactose greater than 4.8%, but this difference was not significant. Of the treated cows with identified bacteria, 52.6% of the quarters were cured, while in the control group only 25.0% (P<0.001). Specifically, all cows identified with Escherichia coli in the treatment group were cured, with 66.6% cured with no intervention in the control. Spontaneous cure of glands infected with coagulase negative staphylococci (CNS) and Streptococci was low while treatment successfully increased the cure of CNS from 13.3% to 53.8% and that of Streptococci from 18.2% to 36.4%. Of the 4 cows identified with Staphylococcus aureus, 3 were cured. The clinical mastitis study group included 29 infected cows that were submitted either to a gold standard antibiotic treatment subgroup of 16 cows (n = 16) or to an APT treatment subgroup of 13 cows (n = 13). A cure of 18.7% was shown for the antibiotic treatment, of which logSCC returned to <5.6 cell/mL and 56.2% were culled. A cure of 76.9% was shown for the APT treatment with only one cow culled (7.7%).

[Cost-benefit analysis of an intervention to improve udder health in Swiss dairy farms]. / Kosten-Nutzen-Analyse einer Intervention zur Verbesserung der Eutergesundheit in Schweizer Milchviehbetrieben.

The objective of this study was to calculate the national costs associated with udder health in Switzerland and to estimate the cost effectiveness of an udder health intervention program. In 49 farms, yearly mastitis associated costs before and during an intervention were collected at herd level. Costs were calculated for each lactating cow being present in the herd. At the beginning of the intervention, 24 farms received a report with recommendations to improve the udder health. In the following year, those herds were followed-up by their veterinarian at a monthly basis. The other 25 farms were used as a negative control group and neither received any recommendations nor any follow-up. In the first year of analysis (2010), the median udder health associated costs were 209.- Swiss Francs for each lactating cow, regardless of the intervention group. During the intervention period (2012), mastitis associated costs were 191.- Swiss Francs for control farms and 396.- Swiss Francs for farms with veterinary intervention on a monthly basis. The median additional costs for herds with intervention were 159.- Swiss Francs per lactating cow. At the national level, mastitis associated costs were estimated at 129.4 millions of Swiss Francs per year. The cost effectiveness of future mastitis control programs can be evaluated with the help of the deterministic model developed during this study.

Le but du présent travail était de calculer les coûts de la santé de la mamelle dans des exploitations laitières suisses et d'estimer l'efficacité économique d'une intervention en vue de l'améliorer. Pour cela on a relevé les coûts liés aux mammites sur une année dans 49 exploitations, et cela durant l'année précédant l'intervention puis dans l'année de l'intervention et on l'a divisé par le nombre de vaches en lactation. Vingt-quatre exploitations ont reçu au début de l'étude des recommandations en vue d'améliorer la santé de la mamelle et ont ensuite été suivies mensuellement durant une année par leur vétérinaire d'exploitation. Les 25 autres exploitations n'ont reçu aucune recommandation et ont été utilisées comme groupe de contrôle. Dans la première année d'analyse (2 ans avant l'intervention, 2010), les coûts moyens de la santé de la mamelle, indépendamment du groupe s'élevaient à CHF 209.­ par vache en lactation. Durant l'année de l'intervention, ils se montaient à CHF 191.­ pour les exploitations de contrôle contre CHF 396.- pour les exploitations suivies. Les dépenses supplémentaires durant l'intervention s'élevaient en moyenne à CHF 159.­ par vache en lactation. Au niveau national, on estime les coûts liés aux mammites à CHF 129.4 millions. Les modèles de calcul utilisés dans la présente étude permettent de juger à l'avenir de la rentabilité des programmes de contrôle des mammites.

Effects of continuous lactation and short dry periods on mammary function and animal health.

The dry period is required to facilitate cell turnover in the bovine mammary gland in order to optimize milk yield in the next lactation. Traditionally, an 8-week dry period has been a standard management practice for dairy cows based on retrospective analyses of milk yields following various dry period lengths. However, as milk production per cow has increased, transitioning cows from the nonlactating state to peak milk yield has grown more problematic. This has prompted new studies on dry period requirements for dairy cows. These studies indicate a clear parity effect on dry period requirement. First parity animals require a 60-day dry period, whereas lactations following later parities demonstrate no negative impact with 30-day dry period or even eliminating the dry period when somatotropin (ST) is also used to maintain milk yields. Shortened dry periods in first parity animals were associated with reduced mammary cell turnover during the dry period and early lactation and increased numbers of senescent cells and reduced functionality of lactating alveolar mammary cells postpartum. Use of ST and increased milking frequency postpartum reduced the impact of shortened dry periods. The majority of new intramammary infections occur during the dry period and persist into the following lactation. There is therefore the possibility of altering mastitis incidence by modifying or eliminating the dry period in older parity animals. As the composition of mammary secretions including immunoglobulins may be reduced when the dry period is reduced or eliminated, there is the possibility that the immune status of cows during the peripartum period is influenced by the length of the dry period.

Management of mastitis on organic and conventional dairy farms.

This paper compares management of mastitis on organic dairy farms with that on conventional dairy farms. National standards for organic production vary by country. In the United States, usage of antimicrobials to treat dairy cattle results in permanent loss of organic status of the animal, effectively limiting treatment choices for animals experiencing bacterial diseases. There are no products approved by the US Food and Drug Administration that can be used for treatment of mastitis on organic dairy farms, and usage of unapproved products is contrary to Food and Drug Administration guidelines. In general, organic dairy farms tend to be smaller, produce less, and more likely to be housed and milked in traditional barns as compared with conventionally managed herds. It is difficult to compare disease rates between herds managed conventionally or organically because perception and detection of disease is influenced by management system. To date, no studies have been published with the defined objective of comparing animal health on organic dairy herds with that on conventional dairy herds in the United States. European studies have not documented significant differences in animal health based on adoption of organic management. Few differences in bulk tank somatic cell counts have been identified between organic and conventional herds. Farmers that have adopted organic management consistently report fewer cases of clinical mastitis, but organic farmers do not use the same criteria to detect clinical mastitis. European dairy farmers that adopt organic management report use of a variety of conventional and alternative therapies for treatment and control of mastitis. In the United States, organic farmers treat clinical mastitis using a variety of alternative therapies including whey-based products, botanicals, vitamin supplements, and homeopathy. Organic farmers in the United States use a variety of alternative products to treat cows at dry-off. Virtually no data are available that support the clinical efficacy of any of the alternative veterinary products used for treatment or prevention of mastitis. Some associations between organic management and antimicrobial susceptibility of gram-positive mastitis pathogens have been noted, but overall, few mastitis pathogens from both conventional and organic dairy herds demonstrate resistance to antibiotics commonly used for mastitis control.

Stochastic modeling to determine the economic effects of blanket, selective, and no dry cow therapy.

In many countries, blanket dry cow therapy (DCT) is the standard way to dry off cows. Because of concerns about antibiotic resistance, selective DCT is proposed as an alternative. The economic consequences of different types of DCT were studied previously, but variation between input traits and different types of pathogens were not taken into account. The goal of this study was to create a stochastic Monte Carlo model to simulate the dynamics of intramammary infections (IMI) around the dry period to predict the economic consequences of DCT for different types of pathogens (Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Staphylococcus aureus, and Escherichia coli). The traits used in the model can be varied. The probabilities within the basic situation were collected from the literature and, because not all information needed was available in the literature, by interviewing experts (n = 10). The expert opinions were translated into minimum, most expected, and maximum values for each of the different probabilities. For Dutch farmers, the costs associated with mastitis and mastitis control around the dry period varied between 10.61 euros and 26.61 euros (average 15.60 euros) for blanket DCT, between 4.86 euros and 29.41 euros (average 13.72 euros) for selective DCT, and between 4.08 euros and 42.60 euros (average 18.02 euros) for no DCT. Although there were small differences between the treatment groups, the variation within the treatment groups was much larger. The major portion of the costs for selective treatment (59% of the total costs) and no DCT (82%) was derived from the costs of clinical mastitis after calving, and for blanket DCT, the costs of treatment (65%) exceeded the costs of clinical mastitis (27%). The cost of mastitis around the dry period was most sensitive to a change in the risk of new IMI during the dry period, spontaneous cure, and costs associated with the antibiotic treatment. The optimal decision to dry off cows depends on the attitude of the farmer toward risk and other farm-specific traits and probabilities such as the new IMI rate during the dry period. Therefore, it is necessary to make farm-specific calculations so that farmers are able to factor this information into their decisions when choosing the best DCT for their situations.

Genetic evaluation of mastitis resistance using a first-passage time model for wiener processes for analysis of time to first treatment.

A Wiener process is a Brownian-motion process initiated in a certain state in a state space, and the first passage time is defined as the time of the process to reach a predefined absorbing state where the process stops. Time from 31 d prepartum to first treatment of clinical mastitis (CM) was modeled as first passage times of such Wiener processes. Two processes were used to allow for several risk factors, and for each process, initiation was at some arbitrary time point, in a certain health state with drift toward or away from absorption (disease). The drift parameter of each process was expressed as linear functions of covariates (year of calving and sire). First passage time was defined as the time from process initiation until the first health status process reached zero (absorption). The model was fitted to records for 36,178 first-lactation daughters of 245 Norwegian cattle sires using a Bayesian approach and Markov chain Monte Carlo methods. Genetic evaluation of sires was carried out by calculating the posterior probability of no CM (the value of the survival function) by d 331, i.e., 300 d after first calving. Alternatively, sire evaluation was based on the integrated area under the survival curve. These measures were highly correlated (0.999), which indicates a small degree of crossings of the sire-dependent survival curves. Hence, sire-specific hazards were close to proportional, resulting in a higher rank-correlation to sire evaluations from a survival model with proportional hazards than to the results from a multivariate threshold model.

Milk loss and treatment costs associated with two treatment protocols for clinical mastitis in dairy cows.

The objective of this study was to compare milk loss and treatment costs for cows with clinical mastitis that were given antibiotics in addition to supportive treatment or supportive treatment alone. Between January 1994 and January 1996, 116,876 daily milk records on 676 lactations were taken at the University of Illinois Dairy Research Farm. Clinical mastitis was diagnosed during 124 lactations with 25,047 daily milk records, and 1417 of the daily milk records were on days when clinical mastitis was present. Cows with clinical mastitis were randomly assigned to one of 2 treatment groups: N (supportive treatment only) or A (antibiotics in addition to supportive treatment). Extent of antibiotic and supportive treatment varied according to twice daily severity scores. Projected and actual daily milk yields were estimated utilizing a random regression test-day model, and the differences were summed over 305 d of lactation to estimate lactational milk yield loss. The actual amount of discarded milk was added to milk yield loss to determine total milk loss per lactation. A cost analysis that included milk loss and treatment costs was then performed. Cows with clinical mastitis that were given only supportive treatment lost 230 +/- 172 kg (mean +/- standard error of mean [SEM]) more milk and incurred 94 +/- 51 dollars (SEM) more cost per lactation than cows given antibiotics and supportive treatment. Cows given only supportive treatment showed a response pattern of 305-d milk yield loss and economic loss per lactation that varied 2 to 3 times as much as cows treated with antibiotics. Based on reduced milk loss, better reliability (less variable response), and lower economic loss, the addition of antibiotics to supportive treatment was more efficacious and cost effective than supportive treatment alone.

Farmers' choice of medical treatment of mastitis in Danish dairy herds based on qualitative research interviews.

A qualitative research study was conducted to describe and analyze farmers' perspectives on their own choices regarding decisions to have cows treated for mastitis. Through qualitative research interviews of 16 Danish dairy farmers, four levels of the decision-making process used by farmers to decide whether or not to treat a cow with antibiotics were identified. Those levels were: 1) symptom level (seriousness of the mastitis case), 2) cow level (to the extent a cow fulfilled goals of the farmer and the herd), 3) herd level (the situation of the herd, e.g., in relation to milk quota), and 4) level of alternatives (whether the farmer regards such practices as blinding of teats or homoeopathy as serious alternatives to antibiotic treatment). All four levels could be recognized in all herds, but with differing weights and relative importance across herds. Directions of different possibilities within each level also varied among farmers. By identifying those four levels, a model for understanding the farmers' choices is provided. This provides background for dialogue with each farmer about choices in the context of each specific herd. It also provides insight into implications of mastitis treatments for effective treatment versus issues of antibiotic resistance when discussing choices on a more general level. Communication and understanding between farmers and their veterinarians and cattle-oriented advisors is essential. Farmers were shown to be coherent in their choices of treatment, but their decisions often seemed to differ from normal veterinary recommendations. Such differences have to be understood and implemented into effective decisions for the whole farm.

Disease management research using event graphs.

Event Graphs, conditional representations of stochastic relationships between discrete events, simulate disease dynamics. In this paper, we demonstrate how Event Graphs, at an appropriate abstraction level, also extend and organize scientific knowledge about diseases. They can identify promising treatment strategies and directions for further research and provide enough detail for testing combinations of new medicines and interventions. Event Graphs can be enriched to incorporate and validate data and test new theories to reflect an expanding dynamic scientific knowledge base and establish performance criteria for the economic viability of new treatments. To illustrate, an Event Graph is developed for mastitis, a costly dairy cattle disease, for which extensive scientific literature exists. With only a modest amount of imagination, the methodology presented here can be seen to apply modeling to any disease, human, plant, or animal. The Event Graph simulation presented here is currently being used in research and in a new veterinary epidemiology course.

The economic benefit of treating subclinical Streptococcus agalactiae mastitis in lactating cows.

The economic benefits of treating lactating cows for Streptococcus agalactiae mastitis were studied at a large (689 milking cows) central California dairy. Postcure milk production of case cows (infected, treated, and cured) was compared with production of paired control cows (uninfected) and was matched for yield, days in milk, days in gestation, and parity. A simulation was used to plot expected lactation curves for mastitic cows (infected, not treated) with characteristics similar to those of each control cow, and these curves were compared with actual case-cow lactation curves. The difference in actual and expected production was used to calculate net economic benefits of treatment. Comparison of expected with actual production indicated a net benefit from treatment of $396/cow for cows treated in early lactation and $237 for cows treated in midlactation, but a net loss of $55 for cows treated in late lactation. Lactation number did not have a significant impact on economic benefits of treatment. In contrast to other studies indicating no economic benefit from treating mastitis during lactation, this study's positive results may have been attributable to the high cure rate (98%) and the subclinical form of mastitis being treated. Streptococcus agalactiae mastitis treatment during early and midlactation would appear to be an economically justifiable option for dairy managers.

Cost benefit analysis of lactation therapy with somatic cell counts as indications for treatment.

Lactating dairy cows (487) from five commercial herds were in a study of benefits from lactation therapy of sub-clinical mastitis. Bacterial isolations and composite milk samples for somatic cell counts were taken from each cow each month for 15 mo. Cows (254) in the experimental group were infused with cephapirin in all quarters for two consecutive milkings if somatic cell counts rose above 400,000 cells/ml; 103 cows were so treated. Stepwise regression showed that lactation number, somatic cell counts, days in milk, and percent quarters infected explained variation in milk production, but treatment group, herd, and season did not. Also, there were no significant differences between production of infected experimental and control cows with high somatic cell counts on test dates after treatment. With the experimental program, there was a net loss of +19.65/cow. Intramammary lactation therapy based on somatic cell counts less than 400,000 cells/ml is not recommended.

[Bovine mastitis: a problem of veterinary medicine, economics and food hygiene (author's transl)]. / Die Rindermastitis als veterinärmedizinisches, wirtschaftliches und lebensmittelhygienisches Problem.

Veterinary medicine, dairying, and consumers are likewise interested in mastitis control. The veterinary point of view includes health and function of the udder as well as the production of milk of hygienic quality. Investigations in 6 dairy farms showed that a mastitis control program improved the udder health and resulted in economic benefits. Up to 67% of the subclinical infected udder quarters could be eliminated by prophylactical control measures, especially teat dipping and dry cow therapy. The frequency of clinical udder infections was reduced by about 50%. Furthermore in the first year 5.1% and 7.1% increase of milk and fat yield respectively could be noticed for cows being subject to the program for the whole time. These values could not be reached in the second year. In one herd the milk production increased by about 10% after a control period of two years. The owner of this herd (ca. 35 cows), who was very engaged in the program, obtained an economical benefit of ca. 3500 DM in the first and 7300 DM in the second year of the program. This realised a profit of1.85 DM in the first year and 3.85 DM in the second year per each DM cost in the control program. Mastitis as well as control measures do not implicate a special risk for the human health, because food and drug regulations enable the protection of the consumer in an optimal manner.