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To design cost-effective prevention strategies against mastitis in dairy cow farms, knowledge about infection pathways of causative pathogens is necessary. Therefore, we investigated the reservoirs of bacterial strains causing intramammary infections in one dairy cow herd. Quarter foremilk samples (n = 8056) and milking- and housing-related samples (n = 251; from drinking troughs, bedding material, walking areas, cow brushes, fly traps, milking liners, and milker gloves), were collected and examined using culture-based methods. Species were identified with MALDI-TOF MS, and selected Staphylococcus and Streptococcus spp. typed with randomly amplified polymorphic DNA-PCR. Staphylococci were isolated from all and streptococci from most investigated locations. However, only for Staphylococcus aureus, matching strain types (n = 2) were isolated from milk and milking-related samples (milking liners and milker gloves). Staphylococcus epidermidis and Staphylococcus haemolyticus showed a large genetic diversity without any matches of strain types from milk and other samples. Streptococcus uberis was the only Streptococcus spp. isolated from milk and milking- or housing-related samples. However, no matching strains were found. This study underlines the importance of measures preventing the spread of Staphylococcus aureus between quarters during milking.
To effectively prevent and control bovine mastitis, farmers and their advisors need to take infection pathways and durations into account. Still, studies exploring both aspects through molecular epidemiology with sampling of entire dairy cow herds over longer periods are scarce. Therefore, quarter foremilk samples were collected at 14-d intervals from all lactating dairy cows (n = 263) over 18 wk in one commercial dairy herd. Quarters were considered infected with Staphylococcus aureus, Streptococcus uberis, or Streptococcus dysgalactiae when ≥100 cfu/mL of the respective pathogen was detected, or with Staphylococcus epidermidis or Staphylococcus haemolyticus when ≥500 cfu/mL of the respective pathogen was detected. All isolates of the mentioned species underwent randomly amplified polymorphic DNA (RAPD)-PCR to explore strain diversity and to distinguish ongoing from new infections. Survival analysis was used to estimate infection durations. Five different strains of Staph. aureus were isolated, and the most prevalent strain caused more than 80% of all Staph. aureus infections (n = 46). In contrast, 46 Staph. epidermidis and 69 Staph. haemolyticus strains were isolated, and none of these caused infections in more than 2 different quarters. The 3 most dominant strains of Strep. dysgalactiae (7 strains) and Strep. uberis (18 strains) caused 81% of 33 and 49% of 37 infections in total, respectively. The estimated median infection duration for Staph. aureus was 80 d, and that for Staph. epidermidis and Staph. haemolyticus was 28 and 22 d, respectively. The probability of remaining infected with Strep. dysgalactiae or Strep. uberis for more than 84 and 70 d was 58.7 and 53.5%, respectively. Staphylococcus epidermidis and Staph. haemolyticus were not transmitted contagiously and the average infection durations were short, which brings into question whether antimicrobial treatment of intramammary infections with these organisms is justified. In contrast, infections with the other 3 pathogens lasted longer and largely originated from contagious transmission.
Although Corynebacterium spp. can be regularly associated with subclinical and clinical mastitis cases in dairy cows, knowledge on their reservoirs in dairy farms is sparse. Therefore, samples were collected at 10 visits with 14 day intervals from bedding material (n = 50), drinking troughs (n = 20), different walking areas (n = 60), cow brushes (n = 8), fly traps (n = 4), the passage to pasture (n = 9) as well as milking liners (n = 80) and milker gloves (n = 20) in one dairy cow farm. Additionally, quarter foremilk samples from all lactating cows (approximately 200) were collected at each visit. All samples underwent microbiological examination and cultured isolates were identified using MALDI-TOF MS. Most Corynebacterium spp. that were cultivated from milk were also isolated from the housing environment and milking-related niches (C. amycolatum, C. confusum, C. stationis, C. variabile, C. xerosis) or from milking-related niches only (C. frankenforstense, C. pilosum, C. suicordis). C. bovis was not cultivated from any environmental niche, while being the dominant species in milk samples. This study demonstrates that many Corynebacterium spp. present in milk samples can also be isolated from the cows’ environment. For C. bovis, the most relevant Corynebacterium species with regard to intramammary infections, it indicates that environmental reservoirs are of little relevance.
Lactational treatment of bovine mastitis is a major contributor to antibiotic consumption in dairy cattle and is, therefore, important to address in light of the increasing problem of antibiotic resistance. In this large-scale database-based retrospective observational study, we combined electronic health records and routinely measured somatic cell counts from individual cows to create an overview of lactational mastitis treatment in Danish dairy herds from 2010 to 2019. Furthermore, posttreatment somatic cell count was used to approximate treatment success in terms of cytological cure. A generalized logistic regression with mixed effects was performed to combine knowledge on cow-level factors (treatment-, pathogen-, and cow-related) with the new infection risk at the herd level, and to explore the relative effect on cytological cure. The investigation revealed that the total number of lactational treatments appears to have decreased steadily over the study period, whereas treatment duration increased slightly. The proportion of cases treated with penicillin-based protocols and the proportion of milk samples sent for pathogen analysis also decreased. Meanwhile, results from the statistical analysis confirm the importance of cow-related factors, such as parity and lactation stage, for the probability of cytological cure following lactational treatment of mastitis. However, they also disclose that factors that are easier to adjust, such as optimizing treatment duration, including knowledge on causative pathogens and improving the herd-level new infection risk that can be used to positively influence the outcome. Application of this knowledge could potentially assist in promoting a more prudent use of antibiotics for dairy cattle in the future.
Bovine mastitis is one of the most important diseases in modern dairy farming, as it leads to reduced welfare and milk production and increased need for antibiotic use. Clinical mastitis in Denmark is most often treated with a combination of local and systemic treatment with penicillin. The objective of this randomized clinical trial was to assess whether worse results could be expected with local intramammary treatment with penicillin compared with a combination of local and systemic treatment with penicillin in terms of the bacteriological cure of mild and moderate clinical mastitis cases caused by gram-positive bacteria. We carried out a noninferiority trial with a noninferiority margin set to a relative reduction in bacteriological cure of 15% between these 2 treatment groups to assess the effect of reducing the total antibiotic use by a factor of 16 for each treated case. Clinical mastitis cases from 12 Danish dairy farms were considered for enrollment. On-farm selection of gram-positive cases was carried out by the farm personnel within the first 24 h after a clinical mastitis case was detected. A single farm used bacterial culture results from the on-farm veterinarian, whereas the other 11 farms were provided with an on-farm test to distinguish gram-positive bacteria from gram-negative or samples without bacterial growth. Cases with suspected gram-positive bacteria were allocated to a treatment group: either local or combination. Bacteriological cure was assessed based on the bacterial species identified in the milk sample from the clinical mastitis case and 2 follow-up samples collected approximately 2 and 3 wk after ended treatment. Identification of bacteria was carried out using MALDI-TOF on bacterial culture growth. Noninferiority was assessed using unadjusted cure rates and adjusted cure rates from a multivariable mixed logistic regression model. Of the 1,972 clinical mastitis cases registered, 345 (18%) met all criteria for inclusion (full data). The data set was further reduced to 265 cases for the multivariable analysis to include only complete registrations. Streptococcus uberis was the most commonly isolated pathogen. Noninferiority was demonstrated for both unadjusted and adjusted cure rates. The unadjusted cure rates were 76.8% and 83.1% for the local and combined treatments, respectively (full data). The pathogen and somatic cell count before the clinical case had an effect on the efficacy of treatment; thus efficient treatment protocols should be herd- and case-specific. The effect of pathogen and somatic cell count on treatment efficacy was similar irrespective of the treatment protocol. We conclude that bacteriological cure of local penicillin treatment for mild and moderate clinical mastitis cases was noninferior to the combination of local and systemic treatment using a 15% noninferiority margin. This suggests that a potential 16-fold reduction in antimicrobial use per mastitis treatment can be achieved with no adverse effect on cure rate.
Mastitis is a major health problem for bovines and can be categorized as non-severe or severe, based on clinical symptoms. A severe case of clinical mastitis is usually defined by the cow being affected systemically. It is important to consider how to handle severe cases because these cases can be fatal and cause high production losses. However, there are generally few detailed treatment guidelines. By conducting a scoping review on the topic, we aimed to synthesize the information that is available on treatment and outcomes, as reported from clinical trials and observational studies. This was facilitated by following the PRISMA-guidelines with a stepwise systematic screening of scientific literature on the subject, retrieved via Pubmed and Web of Science, using pre-defined selection criteria. The results yielded a total of 14 reports of treatment and outcomes in cases of naturally occurring severe clinical mastitis. Cross-trial comparison was difficult due to the different exclusion criteria and outcome definitions. Many studies focused on cases caused by gram-negative bacteria treated with intensive antibiotic protocols, often containing antibiotics that are categorized as critical for human health. Few focused on severe cases caused by gram-positive bacteria or on the relative use of non-antibiotic treatment. In general, only a small number of statistically significant differences were found in trials comparing different treatment protocols, with no obvious trends across trials. Our findings emphasize the need for more research into the treatment efficacy of antibiotic and non-antibiotic options for clinically severe mastitis. Furthermore, consideration of how trial conditions relate to the practical circumstances in a field setting could improve the applicability of reported results. This could help to provide practitioners with the information needed to make evidence-based treatment decisions in cases of clinically severe mastitis.
