Heat stress and cow factors affect bacteria shedding pattern from naturally infected mammary gland quarters in dairy cattle.

Mastitis-causing pathogens are shed from infected mammary gland quarters and thus contribute to an increased risk of new intramammary infections. The objective of the current study was to investigate the shedding characteristics of various mastitis-causing pathogens and associated animal-specific (somatic cell score and parity) and environmental (heat stress) factors. In a longitudinal study, infected udder quarters were sampled consecutively on 5 dairy farms in Germany. To capture climatic factors, temperature-humidity index (THI) was calculated. In the laboratory analysis, the pathogens and their counts in the milk samples were determined. A generalized linear mixed model with gamma link was used to evaluate the factors influencing pathogen-shedding characteristics. The variables somatic cell count, pathogen, parity, and THI had significant influence on pathogen shedding. Staphylococci were shed in lower values than streptococci. The pathogen shedding from mammary gland quarters with intramammary infections was higher in the first and second lactation than in higher lactations. Exceeding the THI threshold 60 resulted in higher pathogen counts on the same day. This was only caused by the pathogens yeasts and Streptococcus uberis. Possible mechanisms causing differences in pathogen shedding are changes in the counts due to influenced milk quantities, better growth conditions at higher temperatures, or altered immunological reactions. The mechanisms often remain speculative and require further investigation. The study underlines the contribution of cows with high somatic cell counts regarding the transmission of mastitis pathogens within a herd. Furthermore, it becomes clear that heat stress in Germany influences udder health and that prevention measures are useful.

Comparison of phenotypic and genotypic antimicrobial resistance patterns associated with Staphylococcus aureus mastitis in German and Danish dairy cows.

Staphylococcus aureus is one of the most common pathogens associated with bovine mastitis in Germany and Denmark. Successful therapy is strongly linked to the susceptibility of the pathogen to the administered antimicrobial. An increase in resistant pathogens in human and veterinary medicine has become a concern worldwide and hampers therapy due to reduced susceptibility. In the present study, susceptibility testing was performed for 85 and 93 S. aureus isolates originating from mastitis cases on 12 German and 8 Danish dairy farms, respectively. Phenotypic examination was performed by detection of minimal inhibitory concentration (MIC) values using the broth microdilution method, followed by genotypic investigations of the blaZ and mecA resistance genes via PCR. The tested antimicrobials were the most frequently used ß-lactams in German and Danish dairy farms, including cefquinome, cefoperazone, cephapirin, penicillin, oxacillin, cloxacillin, amoxicillin-clavulanic acid, and cephalexin-kanamycin. Special attention was paid to varying therapy concepts because, in Germany, third- and fourth-generation cephalosporins have been predominantly used in mastitis therapy, whereas in Denmark, restrictive use of penicillin is followed by a general avoidance of cephalosporins. Differences in MIC values were mainly based on determined MIC90 values (MIC at which 90% of isolates are inhibited). In general, Danish S. aureus isolates were inhibited at comparatively lower MIC90 values than S. aureus isolated from German dairy farms for most ß-lactams. No differences were observed regarding cefquinome, because both German and Danish isolates exhibited MIC50 and MIC90 values of 0.5 and 1 µg/mL, respectively. In contrast, the MIC90 for penicillin against German and Danish S. aureus were 0.5 and ≤0.06 µg/mL, respectively. Resistance genes (blaZ, mecA) were only detected in German S. aureus isolates on 3 dairy farms in Germany. A total of 5 isolates tested positive for both blaZ and mecA, whereas 1 isolate carried the blaZ resistance gene only. A direct correlation between frequently used antimicrobials and reduced susceptibility could not be determined based on results of the present study. In addition to further research to determine factors associated with resistance development, we emphasize the urgent need for internationally standardized clinical breakpoints to assess resistance situations more accurately.

In vitro ability of mastitis causing pathogens to form biofilms.

This Research Communication describes the study of in vitro biofilm formation of mastitis causing pathogens. Biofilms are communities of bacteria that are attached to a surface and to each other and are embedded in a self-produced matrix of extracellular polymeric substances. Biofilm formation is an important virulence factor that may result in recurrent or persistent udder infections and treatment failure through increased resistance to antibiotics and protection against host defences. In the present study 252 bacterial isolates from milk samples from bovine udder quarters with intramammary infections were examined with Congo Red agar (CRA) method and tube method (TM) for their ability to form biofilms. Both tests revealed a high number of biofilm-positive strains. Literature reports that the cure rates for Staphylococcus aureus infected udders are lower (27%) in comparison to cure rates of Streptococcus uberis (64-81%) or coagulase-negative staphylococci (CNS) mastitis (80-90%). The findings of the present study suggest that biofilm formation is not the main factor for the differences in cure rates of the various bacteria genera, because all tested pathogen groups showed a similarly high proportion of biofilm formation. Further research is needed to detect microbial biofilms on bovine udder epithelia.

Selection of cows for treatment at dry-off on organic dairy farms.

Restrictions regarding the use of antibiotics make selective antibiotic dry cow therapy (DCT) mandatory on organic farms in Germany. This requires methods for identifying cows with an intramammary infection (IMI) at dry-off. The aim of this field study was to create a decision scheme for the use of DCT based on cow level factors associated with IMI at dry-off and the probability of both cure and new infection (NI) during the dry period. Data from 250 cows from five organic farms were collected including somatic cell counts (SCC) from Dairy Herd Improvement (DHI) records, California mastitis test (CMT) results at dry-off, clinical mastitis (CM) history, parity and dry-off treatment. IMI at dry-off were most accurate identified using a geometric mean SCC of 100 000 cells/ml as a threshold at either one or three DHI records prior to dry-off. Using a combination of SCC with either CM history, CMT at dry off or parity slightly increased the sensitivity of detection (SE). The probability of cure of the infection over the dry period increased with use of both antibiotic DCT and application of an internal teat sealant (ITS) and decreased when the dry period was longer than 56 d. The risk of NI decreased with the use of ITS and infections with minor pathogens at dry-off. Compared with the selection performed by the farmers during the study period identification of IMI based on the selection criterion with a defined SCC threshold achieved a higher SE.

The Influence of Milk Leakage, Udder Pressure and Further Risk Factors on the Development of New Intramammary Infections during the Dry Period of Dairy Cows.

Prevention of new intramammary infection (NIMI) during the dry period (DP) is essential to prevent the development of mastitis in dairy cows. To investigate risk factors for NIMI, 212 cows, comprising a total of 848 udder quarters, were examined in this study. Quarter milk samples were taken on the day of drying off and 7 ± 3 days after calving. Cow- and quarter-level associated risk factors were assessed at the beginning of the DP and after calving. In total, 7.1% of the udder quarters developed an NIMI between the samplings. Non-aureus staphylococci (40.4%) and Gram-negative pathogens (22.8%) were most frequently the cause of NIMI. The observed milk leakage prevalence was 16.7%, with a peak 24 h after drying off. Simultaneously, the udder pressure peaked 24 h after drying off. A significant correlation between milk yield on the day before drying off and milk leakage could be proven. Cows with quarters leaking milk produced an average milk yield of 28.32 kg on the day before drying off. Generalised linear mixed models and odds ratios were calculated to determine the significant risk factors for NIMI during the DP and early lactation. Quarters leaking milk had 3.4 higher odds for NIMI between the samplings compared to quarters without milk leakage. Quarters from cows with dirty udders had 3.1 higher odds of developing an NIMI between the samplings compared to quarters from cows with clean udders. The results of this study demonstrate the importance of dry cow management before drying off and during the critical period of active involution of the udder tissue.

Bacteremia in Severe Mastitis of Dairy Cows.

The aim of this cross-sectional study was to investigate the occurrence of bacteremia in severe mastitis cases of dairy cows. Milk and corresponding blood samples of 77 cases of severe mastitis were bacteriologically examined. All samples (milk and blood) were incubated aerobically and anaerobically to also investigate the role of obligate anaerobic microorganisms in addition to aerobic microorganisms in severe mastitis. Bacteremia occurred if identical bacterial strains were isolated from milk and blood samples of the same case. In addition, pathogen shedding was examined, and the data of animals and weather were collected to determine associated factors for the occurrence of bacteremia in severe mastitis. If Gram-negative bacteria were detected in milk samples, a Limulus test (detection of endotoxins) was also performed for corresponding blood samples without the growth of Gram-negative bacteria. In 74 cases (96.1%), microbial growth was detected in aerobically incubated milk samples. The most-frequently isolated bacteria in milk samples were Escherichia (E.) coli (48.9%), Streptococcus (S.) spp. (18.1%), and Klebsiella (K.) spp. (16%). Obligatory anaerobic microorganisms were not isolated. In 72 cases (93.5%) of the aerobically examined blood samples, microbial growth was detected. The most-frequently isolated pathogens in blood samples were non-aureus Staphylococci (NaS) (40.6%) and Bacillus spp. (12.3%). The Limulus test was positive for 60.5% of cases, which means a detection of endotoxins in most blood samples without the growth of Gram-negative bacteria. Bacteremia was confirmed in 12 cases (15.5%) for K. pneumoniae (5/12), E. coli (4/12), S. dysgalactiae (2/12), and S. uberis (1/12). The mortality rate (deceased or culled) was 66.6% for cases with bacteremia and 34.1% for cases without bacteremia. High pathogen shedding and high humidity were associated with the occurrence of bacteremia in severe mastitis.

Severity of Clinical Mastitis and Bacterial Shedding.

The aim of this cross-sectional study was to investigate associated factors of the severity of clinical mastitis (CM). Milk samples of 249 cases of CM were microbiologically examined, of which 27.2% were mild, 38.5% moderate, and 34.3% severe mastitis. The samples were incubated aerobically and anaerobically to investigate the role of aerobic and anaerobic microorganisms. In addition, the pathogen shedding was quantitatively examined, and animal individual data, outside temperature and relative humidity, were collected to determine associated factors for the severity of CM. The pathogen isolated the most was Escherichia coli (35.2%), followed by Streptococcus spp. (16.4%). Non-aureus staphylococci (NaS) (15.4%) and other pathogens (e.g., Staphylococcus aureus, coryneforms) (15.4%) were the pathogens that were isolated the most for mild mastitis. Moderate mastitis was mostly caused by E. coli (38%). E. coli was also the most common pathogen in severe mastitis (50.6%), followed by Streptococcus spp. (16.4%), and Klebsiella spp. (10.3%). Obligate anaerobes (Clostridium spp.) were isolated in one case (0.4%) of moderate mastitis. The mortality rate (deceased or culled due to the mastitis in the following two weeks) was 34.5% for severe mastitis, 21.7% for moderate mastitis, and 4.4% for mild mastitis. The overall mortality rate of CM was 21.1%. The pathogen shedding (back logarithmized) was highest for severe mastitis (55,000 cfu/mL) and E. coli (91,200 cfu/mL). High pathogen shedding, low previous somatic cell count (SCC) before mastitis, high outside temperature, and high humidity were associated with severe courses of mastitis.

Reservoirs of Staphylococcus spp. and Streptococcus spp. Associated with Intramammary Infections of Dairy Cows.

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.

Reservoirs of Corynebacterium spp. in the Environment of Dairy Cows.

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.

Intermediate Cluster Disinfection: Which Disinfection Solution Is Most Effective on Milking Liners? A Comparison of Microorganism Reduction on Liner Inner Surfaces Using Quantitative Swab Sampling Technique.

During machine milking, pathogenic microorganisms can be transmitted from cow to cow through liners. Therefore, in Germany, a spray method for the intermediate disinfection of the milking cluster is often used for prevention. This method of cluster disinfection is easy to perform, requires little time and no extra materials, and the disinfection solution is safe from outside contamination in the spray bottle. Since no data on a systematic efficacy trial are available, the aim of this study was to determine the microbial reduction effect of intermediate disinfection. Therefore, laboratory and field trials were conducted. In both trials, two sprays of 0.85 mL per burst of different disinfectant solutions were sprayed into the contaminated liners. For sampling, a quantitative swabbing method using a modified wet-dry swab (WDS) technique based on DIN 10113-1: 1997-07 was applied. Thus, the effectiveness of disinfectants based on Peracetic Acid, Hydrogen Peroxide and Plasma-Activated Buffered Solution (PABS) was compared. In the laboratory trial, the inner surfaces of liners were contaminated with pure cultures of Escherichia (E.) coli, Staphylococcus (S.) aureus, Streptococcus (Sc.) uberis and Sc. agalactiae. The disinfection of the contaminated liners with the disinfectants resulted in a significant reduction in bacteria with values averaging 1 log for E. coli, 0.7 log for S. aureus, 0.7 log for Sc. uberis and 0.8 log for Sc. agalactiae. The highest reduction was obtained for contamination with E. coli (1.3 log) and Sc. uberis (0.8 log) when PABS was applied and for contamination with S. aureus (1.1 log) and Sc. agalactiae (1 log) when Peracetic Acid Solution (PAS) was used. Treatment with sterile water only led to an average reduction of 0.4 log. In the field trial, after the milking of 575 cows, the liners were disinfected and the total microorganism count from the liner surface was performed. The reduction was measured against an untreated liner within the cluster. Although a reduction in microorganisms was achieved in the field trial, it was not significant. When using PAS, a log reduction of 0.3 was achieved; when using PABS, a log reduction of 0.2 was obtained. The difference between the two disinfection methods was also not significant. Treatment with sterile water only led to a reduction of 0.1 log. The results show that spray disinfection under these circumstances does result in a reduction in the bacteria on the milking liner surface, but for effective disinfection a higher reduction would be preferred.

Intramammary infections with Corynebacterium spp. in bovine lactating udder quarters.

Corynebacterium spp. are frequently detected in bovine quarter milk samples, yet their impact on udder health has not been determined completely. In this longitudinal study, we collected quarter milk samples from a dairy herd of approximately 200 cows, ten times at 14 d intervals. Bacteriologically, Catalase-positive and Gram-positive rods were detected in 22.7% of the samples. For further species diagnosis, colonies were analyzed by MALDI-TOF MS. Corynebacterium bovis, C. amycolatum, C. xerosis and 10 other Corynebacterium spp. were detected. The three aforementioned species accounted for 88.4%, 8.65% and 0.94% of all cultured Corynebacterium spp., respectively. For further evaluation of infection dynamics, the following three infection definitions were applied: A (2/3 consecutive samples positive for the same species), B (≥1000 cfu/mL in one sample), C (isolated from a clinical mastitis case). Infections according to definition B occurred most frequently and clinical mastitis with Corynebacterium spp. occurred once during sampling. Life tables were used to determine the duration of infection. According to infection definition A, infection durations of 111 d and 98 d were obtained for C. bovis and C. amycolatum, respectively. Exemplarily, longer lasting infections were examined for their strain diversity by RAPD PCR. A low strain diversity was found in the individual quarters that indicates a longer colonization of the udder parenchyma by C. bovis and C. amycolatum.

Factors Associated with the Severity of Clinical Mastitis.

Severe mastitis can lead to considerable disturbances in the cows' general condition and even to septicemia and death. The aim of this cross-sectional study was to identify factors associated with the severity of the clinical expression of mastitis. Streptococcus (Str.) uberis (29.9%) was the most frequently isolated pathogen, followed by coliform bacteria (22.3%). The majority of all mastitis cases (n = 854) in this study were either mild or moderate, but 21.1% were severe. It can be deduced that the combination of coliform pathogens and increasing pathogen shedding of these showed associations with severe mastitis. Furthermore, animal-related factors associated with severe disease progression were stages of lactation, and previous diseases in the period prior to the mastitis episode. Cows in early lactation had more severe mastitis. Ketosis and uterine diseases in temporal relation to the mastitis were associated with more severe mastitis in the diseased cows. Hypocalcemia was significantly associated with milder mastitis. As another factor, treatment with corticosteroids within two weeks before mastitis was associated with higher severity of mastitis. Knowledge of these risk factors may provide the basis for randomized controlled trials of the exact influence of these on the severity of mastitis.

Cell Differentiation of Bovine Milk Control Samples to Improve Prognosis of Mastitis Cure.

To optimise udder health at the herd level, identifying incurable mastitis cases as well as providing an adequate therapy and culling strategy are necessary. Cows with clinical mastitis should be administered antibiotic medication if it is most likely to improve mammary cure. The somatic cell count (SCC) in milk of the monthly implemented Dairy Herd Improvement (DHI) test represents the most important tool to decide whether a cow has a promising mammary cure rate. Differential cell count (DCC) facilitates the specification of the immunological ability of defence, for example by characterising leukocyte subpopulations or cell viability. The aim of this study was to assess the DCC and cell viability in DHI milk samples regarding the cytological (CC) and bacteriological cure (BC) of the udder within a longitudinal study, thereby gaining a predictive evaluation of whether a clinical mastitis benefits from an antibiotic treatment or not. The cows enrolled in this study had an SCC above 200,000 cells/mL in the previous DHI test. Study 1 assessed the CC by reference to the SCC of two consecutive DHI tests and included 1010 milk samples: 28.4% of the mammary glands were classified as cytologically cured and 71.6% as uncured. The final mixed logistic regression model identified the total number of non-vital cells as a significant factor associated with CC. An increasing amount of non-vital cells was related to a lower individual ability for CC. Cows which were in the first or second lactation possessed a higher probability of CC than cows having a lactation number above two. If animals developed a clinical mastitis after flow cytometric investigation, the BC was examined in study 2 by analysing quarter foremilk samples microbiologically. Taking 48 milk samples, 81.3% of the mammary glands were classified as bacteriologically cured and 18.7% as uncured. The percentage of total non-vital cells tended to be lower for cows which were cured, but no significance could be observed. This study revealed that the investigation of the proportion of non-vital cells in DHI milk samples can enhance the prognosis of whether an antibiotic treatment of clinical mastitis might be promising or not. Prospectively, this tool may be integrated in the DHI tests to facilitate the decision between therapy or culling.

Investigations on Transfer of Pathogens between Foster Cows and Calves during the Suckling Period.

To date, there have been few studies on the health effects of foster cow systems, including the transmission of mastitis-associated pathogens during suckling. The present study aimed to compare the pathogens detected in the mammary glands of the foster cow with those in the oral cavities of the associated foster calves and to evaluate the resulting consequences for udder health, calf health and internal biosecurity. Quarter milk sampling of 99 foster cows from an organic dairy farm was conducted twice during the foster period. Oral cavity swabs were taken from 345 foster calves. Furthermore, quarter milk samples were collected from 124 biological dams to investigate possible transmission to the foster cows via the suckling calves. All samples were microbiologically examined and confirmed by MALDI-TOF (matrix-assisted laser desorption time-of-flight mass-spectrometry). Using RAPD-PCR (randomly amplified polymorphic DNA polymerase chain reaction), strain similarities were detected for Pasteurella multocida, Staphylococcus aureus, S. sciuri and Streptococcus (Sc.) suis. Transmission of P. multocida and S. aureus probably occurred during suckling. For S. sciuri and Sc. suis, environmental origins were assumed. Transmission from dam to foster cow with the suckling calf as vector could not be clearly demonstrated.

Are Severe Mastitis Cases in Dairy Cows Associated with Bacteremia?

The objective of this study was to investigate the occurrence of bacteremia in dairy cows with severe mastitis. Milk samples were collected from affected udder quarters, and corresponding blood samples were collected from dairy cows with severe mastitis at the time of diagnosis before any therapeutic measures were undertaken. The cultural detection of pathogens in blood classified a bacteremia. Further diagnostic tests were performed to provide evidence of bacteremia. This was realized by PCR with regard to S. aureus, E. coli and S. uberis and the Limulus test. Detection of culturable pathogens in the blood of cows with severe clinical mastitis was rare and occurred in only one of 70 (1.4%) cases. Overall, bacterial growth was detected in 53 of 70 (75.7%) milk samples. S. uberis (22/70), E. coli (12/70) and S. aureus (4/70) were the most frequently isolated pathogens from milk of cows with severe mastitis. PCR was performed in 38 of 70 (54.3%) blood samples. PCR was positive in eight of 38 cases. S. uberis was found most frequently in six blood samples (8.6%). E. coli was found on PCR in one blood sample (1.4%). S. aureus was identified in one blood sample (1.4%). When Coliforms were detected in the quarter milk sample, a Limulus test was performed in the corresponding blood sample. In three of 15 cases, the Limulus test was positive (4.3% of samples). Further studies are needed to investigate the occurrence of bacteremia in cows with severe mastitis in a higher population size.

Corynebacteria in Bovine Quarter Milk Samples-Species and Somatic Cell Counts.

In this species differentiation study of Corynebacterium spp. (C. spp.), quarter foremilk samples from 48 farms were included. These were obtained from both clinically healthy cows and those with clinical mastitis. First, all samples were examined cyto-microbiologically and all catalase-positive rods were differentiated using the direct transfer method in MALDI-TOF MS. C. bovis, C. amycolatum, C. xerosis, and five other species were identified with proportions of 90.1%, 7.7%, and 0.8% for the named species, respectively, and 1.4% for the remaining unnamed species. In addition, somatic cell count (SCC) was determined by flow cytometry. Based on this, the isolates were classified into four udder health groups: "latent infection", "subclinical mastitis", "clinical mastitis" and "others". Approximately 90% of isolates of C. bovis and C. amycolatum were from latently and subclinically infected quarters. Of the C. bovis isolates, 5.8% were obtained from milk samples from clinical mastitis, whereas C. amycolatum was not present in clinical mastitis. The distribution of groups in these two species differed significantly. The geometric mean SCC of all species combined was 76,000 SCC/mL, almost the same as the SCC of C. bovis. With 50,000 SCC/mL, the SCC of C. amycolatum was slightly below the SCC of C. bovis. Through the species-level detection and consideration of SCC performed here, it is apparent that individual species differ in terms of their pathogenicity. Overall, their classification as minor pathogens with an SCC increase is confirmed.

Dry Period or Early Lactation-Time of Onset and Associated Risk Factors for Intramammary Infections in Dairy Cows.

The aim of this study was to define the time-related period of intramammary infections and its relation to risk factors for intramammary infections and clinical mastitis at cow and quarter levels. In total, 269 German Holstein Frisian dairy cows on three farms in Northern and Eastern Germany were included in this study. Quarter milk samples were collected at dry-off, 3 ± 1 days after calving and 17 ± 3 days after calving, for cytomicrobiological examination. Risk factors at quarter- and cow-level associated with intramammary infections and clinical mastitis were recorded during the trial period. Data were analyzed using logistic regression procedures and odds ratios were calculated. Calving for the second time increased the odds of clinical mastitis during the first 100 days of lactation compared to cows calving for the third time or more. A high milk yield after calving was a risk factor for new infections, with environmental pathogens 17 ± 3 days postpartum. A body condition score after calving less than 3.5 was associated with a decreased risk of having an intra-mammary infection (IMI) with non-aureus staphylococci and coryneforms 3 ± 1 days postpartum and consistent body condition between dry-off and early lactation decreased the risk of intramammary infections after calving. The absence of a ring of hyperkeratosis at the teat apex shown at dry-off was associated with a lower risk of intramammary infections with environmental pathogens 17 ± 3 days postpartum. This study shows the important influence of the dry period and early lactation on intramammary infections and clinical mastitis postpartum in dairy cows. Udder quarters may have eliminated pathogens during the dry period in 43.6% of cases in this study. Additionally, new infections occurred during early lactation, so 5.1% more quarters were infected 17 ± 3 days compared to 3 ± 1 days postpartum. New infections can be traced to non-aureus staphylococci and Staphylococcus aureus from dry-off up until 3 ± 1 days postpartum, and to non-aureus staphylococci, Staphylococcus aureus and Streptococcus uberis, after calving. In total, 88.7% of the infected quarters showed new infections with another pathogen species 3 ± 1 days postpartum than at dry-off, and 89.2% of the quarters 17 ± 3 days postpartum than 3 ± 1 days postpartum. In conclusion, the early lactation has just as important an influence on intramammary infections postpartum in dairy cows as the dry period. There is the possibility that udder quarters eliminate pathogens during the early lactation, especially during the dry period. However, there is also the danger that new infections manifest, with a large proportion of new infections occurring after calving. Thus, additional control strategies are of great importance to prevent new infections occurring during early lactation as well as during the dry period to reduce negative effects on milk yield and culling hazards in dairy cows by minimizing the associated risk factors.

Non-inferiority Trial Investigating the Efficacy of Non-steroidal Anti-inflammatory Drugs and Antimicrobial Treatment of Mild to Moderate Clinical Mastitis in Dairy Cows With Long-lasting Udder Diseases.

To reduce ineffective antimicrobial usage in the treatment of non-severe clinical mastitis (CM) in cows with long-lasting udder diseases, non-antibiotic therapy with a non-steroidal anti-inflammatory drug (NSAID) was conducted and evaluated in a non-blinded, positively controlled, non-inferiority trial. Therefore, three-time systemic ketoprofen treatment at intervals of 24 h was evaluated in comparison with the reference treatment of solely antibiotic therapy in a field study on nine free-stall dairy farms located in Northern Germany. Cows with previous CM cases in current lactation and/or with long-lasting high somatic cell counts in preceding dairy herd improvement test days were randomly allocated to one of the two treatment groups in cases of mild to moderate CM. Quarter foremilk samples of the affected quarters were taken for cyto-bacteriological investigation before treatment as well as ~14 and 21 d after termination of therapy. Both treatment groups were compared regarding the bacteriological cure (BC) as the primary outcome. Clinical cure (CC) and no CM relapse within 60 d after the end of treatment (no R60) were chosen as secondary outcomes. The study resulted in the following outcomes: Streptococcus uberis was most frequently identified in microbiological culture from pre-treatment samples, followed by Staphylococcus aureus and Escherichia coli and other coliforms. No significant differences between the NSAID treatment and the reference treatment were detected regarding CC and CM recurrence (no R60). Although the sole ketoprofen therapy resulted in a numerically lower likelihood of BC, there were no significant differences to the reference treatment. Considering the selection criteria in this study, the results indicate that in mild to moderate CM cases exclusive treatment with ketoprofen may constitute an alternative to antimicrobial intramammary therapy, providing an opportunity for reduction of antibiotic usage. However, non-inferiority evaluations were inconclusive. Further investigations with a larger sample size are required to confirm the results and to make a distinct statement on non-inferiority.

Streptococcus dysgalactiae-Contagious or Environmental?

Streptococcus dysgalactiae is among the most important pathogens causing bovine mastitis. Unfortunately, there is presently a lack of clear knowledge about the mode of transmission-contagious or environmental-of this pathogen. To obtain more information on this, knowledge of the genetic diversity of the isolated microorganisms at the farm level can be useful. To observe the strain variety in different herds of cattle, isolates of Strep. dysgalactiae were collected from clinical mastitis samples at different farms, and the strains were typed using the pulsed-field gel electrophoresis (PFGE) method. Overall, we performed strain typing on 93 isolates from 16 farms in Germany and used an index to describe the degree of contagiosity of Strep. dysgalactiae at each farm. This index (CI) represents the number of isolates divided by the number of strains found in mastitis milk of clinical cases within a period of 14 months. The results differed between the farms. In one farm, all six Strep. dysgalactiae cases that occurred during the study period were caused by a single strain (CI = 6), while in another farm the six cases that occurred were caused by five different strains (CI = 1.2). All other farms fell between these two extremes. This indicates that Strep. dysgalactiae infections can occur via several routes of transmission. At the farm level, strain comparisons are necessary to determine the routes of transmission. Two strains were able to survive on the farm for a minimum of 14 months.

In Vitro Susceptibility of Mastitis Pathogens Isolated from Clinical Mastitis Cases on Northern German Dairy Farms.

The present research study investigated the susceptibility of common mastitis pathogens-obtained from clinical mastitis cases on 58 Northern German dairy farms-to routinely used antimicrobials. The broth microdilution method was used for detecting the Minimal Inhibitory Concentration (MIC) of Streptococcus agalactiae (n = 51), Streptococcus dysgalactiae (n = 54), Streptococcus uberis (n = 50), Staphylococcus aureus (n = 85), non-aureus staphylococci (n = 88), Escherichia coli (n = 54) and Klebsiella species (n = 52). Streptococci and staphylococci were tested against cefquinome, cefoperazone, cephapirin, penicillin, oxacillin, cloxacillin, amoxicillin/clavulanic acid and cefalexin/kanamycin. Besides cefquinome and amoxicillin/clavulanic acid, Gram-negative pathogens were examined for their susceptibility to marbofloxacin and sulfamethoxazole/trimethoprim. The examined S. dysgalactiae isolates exhibited the comparatively lowest MICs. S. uberis and S. agalactiae were inhibited at higher amoxicillin/clavulanic acid and cephapirin concentration levels, whereas S. uberis isolates additionally exhibited elevated cefquinome MICs. Most Gram-positive mastitis pathogens were inhibited at higher cloxacillin than oxacillin concentrations. The MICs of Gram-negative pathogens were higher than previously reported, whereby 7.4%, 5.6% and 11.1% of E. coli isolates had MICs above the highest concentrations tested for cefquinome, marbofloxacin and sulfamethoxazole/trimethoprim, respectively. Individual isolates showed MICs at comparatively higher concentrations, leading to the hypothesis that a certain amount of mastitis pathogens on German dairy farms might be resistant to frequently used antimicrobials.