Intramammary Infections In Cows Research Articles

Effects of bovine casein hydrolysate as a dry cow therapy on prevention and cure of bovine intramammary infection, milk production, and somatic cell count in the subsequent lactation

The primary objectives were to investigate the efficacy of bovine casein hydrolysate (bCNH) as a dry cow therapy at (1) preventing new intramammary infection (IMI) postpartum of all bacteria and coagulase-negative staphylococci (CNS), and (2) curing existing subclinical infections, mainly of CNS. The secondary objective was to measure the effects of bCNH on milk yield, composition, and somatic cell count (SCC) during the lactation postcalving. The trial was conducted as a randomized, blinded controlled experiment. Israeli Holstein dairy cows (n = 170) in first or higher lactations were recruited from 4 large commercial dairy herds. Cows were enrolled following clinical examination and bacteriological sampling of each quarter, which was the experimental unit. Random allocation was implemented at the cow level. All quarters of 100 cows were treated with 1 dose of bCNH (60 mg diluted in 20 mL of sterile solution) and those of 70 control cows were treated with saline solution. Clinical assessment of each cow's general appearance, teat-end leakage, and teat morphology was performed for 0, 1, 2, 3, 7, and 14 d after treatment, together with follow-up clinical observation and clinical examination of udder quarters. Quarter aseptic milk samples were obtained for bacteriological culture 48 h pretreatment, at time of treatment, and 3 and 5 d postcalving. Multivariable analyses were conducted to study the effects of bCNH on cure and prevention of IMI, adjusting for parity, farm, average of daily milk yield for 305 d, and average of monthly SCC values for 305 d of previous lactation. The odds of preventing IMI in cows treated with bCNH at dry-off were 2.15 times higher [95% confidence interval (CI): 1.15 to 4.00] than in cows treated with saline. Prevention was mostly of CNS. The odds of preventing CNS in cows treated with bCNH at dry-off were 2.20 times higher (95% CI: 1.58 to 3.07) than in control cows. The odds of curing IMI caused by CNS in cows treated with bCNH at dry-off were 4.80 times higher (95% CI: 0.75 to 30.75) than in saline-treated cows. Log SCC, adjusted to that of the previous lactation, was lower in the bCNH group compared with controls for 305 d in milk postcalving. The average milk yield per day for 305 d, adjusted to average daily milk yield of previous lactation, was higher by 2.1 kg in the bCNH group compared with controls (95% CI: 1.21 to 3.20). Clinical assessment of udders and cows posttreatment showed no negative effects of bCNH. Incidence of stillbirth, clinical mastitis, retained placenta, endometritis (5 to 12 d postcalving), ketosis, abortions, and reproduction did not differ between the 2 groups. Results suggest that a single intramammary administration of bCNH at dry-off effectively increases milk yield and lowers SCC, prevents new IMI during the dry period, and may be a beneficial alternative for curing existing IMI at dry-off, mainly by CNS.

Antimicrobial susceptibility profile of Streptococcus agalactiae strains isolated from bovine mastitis

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.

Total and Differential Cell Counts as a Tool to Identify Intramammary Infections in Cows after Calving.

Simple SummaryMastitis is a costly disease and needs to be identified as soon as possible to reduce the negative effect on milk quality and quantity and to maximize the chance of cure when an antimicrobial therapy is applied. Bacteriological diagnosis is expensive and not easily available in some areas, therefore approaches to reduce the number of samples to be taken, focusing the interest on cows with higher chances to have an intramammary infections are desirable. The results of our study based on a large database of quarter milk samples analyses including bacteriological analysis, total (SCC) and differential (DSCC) cell count in the first 5–30 days after calving suggest a new and sustainable approach. Indeed, a marker (PLCC) calculated by multiplying SCC and DSCC showed to have the lowest cost when applied to identify udder quarters at risk to have an intramammary infection due to major pathogens. Moreover, this approach as well as the one based on SCC became a benefit when the prevalence of these infections exceeds 10%, and it be of high interest, when selective dry cow therapy is applied, to improve animal health at the herd level.Milk differential somatic cells count (DSCC), made possible under field conditions by the recent availability of a high-throughput milk analyzer may represent an improvement in mastitis diagnosis. While an increasing number of studies reports data on DSCC on individual cow samples, very few concerns DSCC from quarter milk samples. This paper reports for the first time the results of a retrospective study aiming to assess the performance of total (SCC), DSCC, and a novel calculated marker (PLCC) measured on quarter milk samples as a method to identify cows at risk for intramammary infection (IMI) in the first 30 days after calving. Overall, 14,586 valid quarter milk samples (3658 cows) taken in the first 30 days of lactation were considered. Quarters with major pathogens (MP) IMI, as expected, showed significantly higher means for SCC, DSCC, and PLCC. The accuracy, sensitivity, and specificity of the diagnosis based on different cut-offs calculated by ROC analysis are relatively close among DSCC, PLCC, and SCC (up to cut-off of 200,000 cells/mL). However, decision-tree analysis which includes the costs of analysis, but also the costs of the actions taken after test results showed as PLCC has the lowest cost among the three markers, and PLCC and SCC are cost effective when MP prevalence is higher than 6–10%. This diagnostic approach is of high interest particularly when selective dry cow therapy is applied to improve animal health at the herd level.

A cross sectional study of prevalence and risk factors associated with subclinical mastitis and intramammary infections, in dairy herds linked to milk collection centers in Rwanda

The objective of this cross-sectional study was to investigate prevalence, causative udder pathogens and their antimicrobial resistance (AMR), as well as cow and herd risk factors associated with subclinical mastitis (SCM = cows with at least one udder quarter with california mastitis test (CMT) score > 2) and intramammary infections (IMI) caused by Staphylococcus(S.) aureus or Non aureus staphylococci (NAS) in dairy cows linked to Milk Collection Centers (MCCs) in Rwanda. Screening for SCM with the CMT was done on 572 cows from 404 herds linked to two MCCs in each of four provinces. Milk from udder quarters with a CMT score ≥3 (scale 1–5) was sampled for bacteriological analysis. Antimicrobial resistance was evaluated in 60 selected S. aureus isolates. Multivariable mixed effect and ordinary logistic regression analyses were performed to identify cow and herd level risk factors associated with SCM, NAS or S. aureus IMI in cows. The prevalence of SCM was 37.3 % at quarter level and 62.0 % at cow level. Bacteria were isolated 73.7 % of the cultured milk samples, whereas 23.3 % were culture-negative and 3.0 % were contaminated. Staphylococcus aureus and NAS were the most prevalent pathogens, representing more than half of all bacteriological findings. Staphylococcus chromogenes and S. epidermidis were the most prevalent NAS species identified. Of the S. aureus strains 83.3 % were resistant for penicillin, 100 % for clindamycin and 20 % tetracycline. The risk factor analysis showed that an increased stage of lactation, dirty udder and legs in single cow herds and lack of calf suckling the dam, dirty udder and legs and lack of feeding cow after milking in multiple cow herds were significantly (P < 0.05) associated with an increased odds of SCM. Similarly, increasing stage of lactation in single cow herds, and housing cows in individual cattle kraal or on earthen floor and hand washing between cows during milking in multiple cow herds were associated with increased odds for NAS IMI. Poor hygiene of milking area in single cow herds and absence of foremilk stripping in multiple cow herds were significantly (P < 0.05) associated with increased odds for S. aureus IMI in cows. In conclusion, SCM prevalence was high across MCCs. The majority of identified pathogens were contagious in nature and they exhibited resistance to penicillin. Control of the identified risks factors and improved biosecurity through adoption of best practices, and farmer training could contribute to lowering SCM prevalence in Rwanda.

Efficacy of novel antiseptic product containing essential oil of Lippia origanoides to reduce intramammary infections in cows.

Background and Aim:The use of antimicrobials in the control of mastitis is of concern in public health due to their inefficiency in targeting microorganisms. Studies with medicinal plants have risen as an alternative to the use of conventional products. The objective of this study was to evaluate the efficacy of an experimental disinfectant based on the essential oil (EO) from Lippia origanoides in preventing the development of new intramammary infections (IMI) in Holstein cows.Materials and Methods:The conventional protocol of pre- and post-milking was used and the control (Conventional treatment [CNV]) and experimental (Experimental treatment [PEX]) products containing EO at 120 μL/mL were applied by immersion. Individual milk samples were analyzed using sheep blood agar methodologies and biochemical tests. The efficiency of the treatment was defined by the presence or absence of Staphylococcus aureus, coagulase-negative Staphylococcus, and Streptococcus spp.Results:There were no clinical and subclinical mastitis cases, no lesions in the mucosal of teats, nor dirt score between groups in this study. Both treatments did not influence the occurrence of IMI.Conclusion:The results revealed that PEX acts efficiently against microorganisms compared to the disinfection by the conventional product demonstrating the efficacy of the alternative product on the prevention of new IMIs in dairy cows.

Differential somatic cell count as an additional indicator for intramammary infections in dairy cows

Mastitis, often caused by intramammary infection (IMI), is a significant problem in dairy farming globally. Somatic cell count (SCC) is widely used as a parameter for screening IMI in cows that are then treated or culled. We investigated the potential of a new parameter, differential SCC (DSCC), to detect IMI at cow level when SCC is already known. We achieved this using bacterial culture (BC) and PCR to detect 4 categories of pathogens (major, minor, other, and any) in 2 Danish dairy herds. Quarter milk samples were collected from monthly dairy herd improvement samplings over 1 yr and analyzed with BC, whereas cow-level dairy herd improvement samples were analyzed using PCR. Days in milk, parity, and IMI status had a significant effect on DSCC. Using DSCC in addition to SCC significantly improved the indication of IMI compared with using only SCC in the any pathogen category in both herds as well as the minor pathogens category in herd 2 when BC was used for detection. When PCR was used to detect IMI, the use of DSCC in addition to SCC was significant for the other pathogens category in herd 1 and the minor pathogens category in herd 2. Thus, our data revealed that DSCC can add significant information describing IMI status even when SCC is already known; however, this depends on the causative pathogen. Future studies may address how to use DSCC in practice as well as consider the availability of temporal data to potentially gain insight into the course of infection.

Cross-sectional study of the relationships among bedding materials, bedding bacteria counts, and intramammary infection in late-lactation dairy cows

Objectives of this study were to (1) describe the intramammary infection (IMI) prevalence and pathogen profiles in quarters of cows approaching dry-off in US dairy herds, (2) compare IMI prevalence in quarters of cows exposed to different bedding material types, and (3) identify associations between bedding bacteria count and IMI in cows approaching dry-off. Eighty herds using 1 of 4 common bedding materials (manure solids, organic non-manure, new sand, and recycled sand) were recruited in a multi-site cross-sectional study. Each herd was visited twice for sampling. At each visit, aseptic quarter-milk samples were collected from 20 cows approaching dry-off (>180 d pregnant). Samples of unused and used bedding were also collected. Aerobic culture was used to determine the IMI status of 10,448 quarters and to enumerate counts (log10 cfu/mL) of all bacteria, Staphylococcus spp., Streptococcus spp. and Streptococcus-like organisms (SSLO), coliforms, Klebsiella spp., noncoliform gram-negatives, Bacillus spp., and Prototheca spp. in unused (n = 148) and used (n = 150) bedding. The association between bedding bacteria count and IMI was determined using multivariable logistic regression with mixed effects. Quarter-level prevalence of IMI was 21.1%, which was primarily caused by non-aureus Staphylococcus spp. (11.4%) and SSLO (5.6%). Only modest differences in IMI prevalence were observed between the 4 common bedding material types. Counts of all bacteria in unused bedding was positively associated with odds of IMI caused by any pathogen [ALL-IMI; odds ratio (OR) = 1.08]. A positive association was also observed for counts of SSLO in unused bedding and SSLO-IMI (OR = 1.09). These patterns of association were generally consistent across the 4 common bedding materials. In contrast, the association between counts of all bacteria in used bedding and ALL-IMI varied by bedding type, with positive associations observed in quarters exposed to manure solids (OR = 2.29) and organic non-manure (OR = 1.51) and a negative association in quarters exposed to new sand (OR = 0.47). Findings from this study suggest that quarter-level IMI prevalence in late-lactation cows is low in US dairy herds. Furthermore, bedding material type may not be an important risk factor for IMI in late lactation. Higher levels of bacteria in bedding may increase IMI prevalence at dry-off in general, but this relationship is likely to vary according to bedding material type.

On-Farm Surfaces in Contact with Milk: The Role of Staphylococcus aureus-Containing Biofilms for Udder Health and Milk Quality.

Staphylococcus aureus is a Gram-positive bacterium that causes intramammary infections and bulk tank milk (BTM) contamination in dairy operations around the world in spite of on-farm application of preventive measures. The study was conducted on a 30-cow dairy farm in the Ñuble Region of Chile. For BTM culture and somatic cell count (SCC) analysis, three consecutive BTM samples were collected. Samples for bacterial culture (n = 16) were collected from macroscopic adherence on previously washed, sanitized, and dry milking equipment surfaces in direct contact with milk during milking or cooling. A total of 48 S. aureus isolates from BTM, milking equipment, and cows' quarters with intramammary infections were analyzed by pulsed-field gel electrophoresis (PFGE). Selected milking equipment pieces were removed for biofilm visualization using scanning electron microscopy (SEM). S. aureus was isolated from all three BTM samples; the average SCC for the three BTM samples was 1,429,333 cells/mL. Fourteen of the 16 samples of milking equipment (87.5%) were culture positive for S. aureus. Biofilms were visualized by SEM in all four removed milking equipment pieces. Microorganisms observed by SEM in those biofilms were mainly coccus-shaped bacteria, and microbiological culture of these biofilms yielded viable S. aureus isolates in all samples. All pulsotypes observed among S. aureus isolates from BTM were indistinguishable from those in milking equipment surfaces. All PFGE pulsotypes observed among S. aureus isolates from biofilms on rubber liners were indistinguishable from isolates from intramammary infections in cows. Our findings suggest that milking equipment films may act as source of S. aureus contamination for BTM and cows during milking, thus compromising the microbiological quality of milk used for manufacturing dairy products.

Signaling pathway underlying splenocytes activation by polysaccharides from Atractylodis macrocephalae Koidz.

Previous study demonstrated that total polysaccharides isolated from Atractylodis macrocephalae Koidz. (RAMPtp) were effective to eliminate intramammary infection in cows. The present study was designed to investigate the immunomodulatory activity of RAMPtp in mouse splenocytes. Splenocyte proliferation, natural killer (NK) cytotoxicity, productions of NO and cytokines, transcription factor activity as well as the signal pathways and receptor were examined. The results showed that RAMPtp significantly promoted splenocyte proliferation and made the cells enter S and G2/M phases, increased ratios of T/B cells, boosted NK cytotoxicity, enhanced transcriptional activities of nuclear factor of activated T cells (NFAT), nuclear factor κB (NF-κB) and activator protein 1 (AP-1), and stimulated secretions of NO, immunoglobulin G (IgG) and multiple cytokine families (IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-6, IL-10, IL-12p40, IL-12p70, IL-13, IFN-γ, TNF-α, G-CSF, GM-CSF, KC, MIP-1α, MIP-1β, RANTES and Eotaxin). In addition, all the specific inhibitors against the mitogen-activated protein kinases (MAPKs) and NF-κB significantly suppressed the IL-6 production induced by RAMPtp. Moreover, splenocytes from Toll-like receptor 4 (TLR4) deficient mouse responded equally to RAMPtp stimulation as the wild-type. Therefore, RAMPtp might induce splenocytes activation at least in part via the TLR4-independent MAPKs and NF-κB signaling pathways. The present results would be useful to further understand the immunomodulatory mechanisms of RAMPtp in elimination of intramammary infection in cows.

Methicillin-resistant Staphylococcus aureus CC22-MRSA-IV as an agent of dairy cow intramammary infections

Methicillin-resistant S. aureus (MRSA) lineages have become major responsible of healthcare- and community-associated infections in human population. Bovine MRSA are sporadically detected in the dairy herd, but its presence enhances the risk of zoonosis. Some lineages are able to lose the specific host tropism, being easily transmitted from animals to humans and vice-versa. The present study aims at clarifying the epidemiology of MRSA intramammary infections in a closed dairy herd, which was running a mastitis control program since years. Quarter milk samples were collected from all lactating cows once a week for 9 weeks and bacteriologically tested. At the end of the follow-up period, also a self-taken nasal swab of the milker was analysed. Three cows (12.5%) were MRSA positive, a fourth showed a transient infection and MRSA was isolated also from the milker’s nose. Somatic cell counts of infected quarters fluctuated from 1000 to 1,800,000 cells/mL. The isolates were genotyped using DNA microarrays and identified as the epidemic UK-EMRSA-15 grouping in CC22. All strains carried the genes for β-lactam and macrolide resistance. The milker isolate differed from cow isolates mainly for the absence of the untruncated β-haemolysin and the presence of the immune evasion cluster. The milker had been volunteering in a nursing home since months, thus playing the role of MRSA vector into the herd. Our results showed the adaptive capacity of such MRSA to the bovine host. Therefore, we suggest that CC22-MRSA should be regarded as a potential cause of reverse zoonosis in dairy cattle herds.

Effect of heat stress on the interaction of Streptococcus uberis with bovine mammary epithelial cells.

Heat stress (HS) negatively affects milk production and has been associated with decreased immune function, and increased rate of intramammary infections (IMI). Research has shown that HS affects gene expression, cell cycle, and cell metabolism in bovine mammary epithelial cells (BMEC). Since BMEC are an initial target of mastitis pathogens, we studied adherence to and internalisation of S. uberis into HS-BMEC, as well as the effect that this interaction has on host cells by measuring HS-BMEC viability and membrane integrity. Results reported in this Research Communication showed that HS reduced cell viability and induced membrane damage. However, these pathological changes, as well as the rate of adherence and internalisation of S. uberis into BMEC, were augmented when S. uberis was cocultured with HS-BMEC. These results may help to understand the pathogenesis of S. uberis IMI as well as the increased susceptibility of mammary glands to IMI in cows subjected to HS.

Association of California Mastitis Test Scores with Intramammary Infection Status in Lactating Dairy Cows Admitted to a Veterinary Teaching Hospital.

BackgroundSubclinical mastitis is of concern in veterinary hospitals because contagious mastitis pathogens might be unknowingly transmitted to susceptible cows and then back to their farm of origin.ObjectivesTo evaluate the California mastitis test (CMT) as an indicator of intramammary infection (IMI) in lactating dairy cows admitted to a veterinary hospital.AnimalsA total of 139 admissions of 128 lactating dairy cows admitted to the University of Illinois Veterinary Teaching Hospital over a 2‐year period.MethodsA retrospective study with a convenience sample was conducted. Medical records of cows with CMT results and milk culture results for the day of admission were reviewed. Breed, age, season, maximum CMT score for the 4 quarters, maximum CMT score difference, and clinical diagnosis were evaluated as predictors of IMI by the chi‐square test and stepwise logistic regression.ResultsAn IMI was identified in 51% of quarters. For cows admitted without evidence of clinical mastitis, the sensitivity of a CMT score ≥trace in predicting an IMI on a quarter or cow basis was 0.45 and 0.68, respectively. The distributions of maximal quarter CMT score and the maximum difference in quarter CMT score for cows without evidence of clinical mastitis did not differ (P = 0.28, P = 0.84, respectively) for cows with and without IMI. Stepwise logistic regression did not identify significant predictors of IMI in cows without clinical mastitis.ConclusionsLactating dairy cattle admitted to a veterinary hospital should be managed as if they have an IMI, even in the absence of clinical mastitis.

Virulence Genes of S. aureus from Dairy Cow Mastitis and Contagiousness Risk.

Staphylococcus aureus (S. aureus) is a major agent of dairy cow intramammary infections: the different prevalences of mastitis reported might be related to a combination of S. aureus virulence factors beyond host factors. The present study considered 169 isolates from different Italian dairy herds that were classified into four groups based on the prevalence of S. aureus infection at the first testing: low prevalence (LP), medium–low (MLP), medium–high (MHP) and high (HP). We aimed to correlate the presence of virulence genes with the prevalence of intramammary infections in order to develop new strategies for the control of S. aureus mastitis. Microarray data were statistically evaluated using binary logistic regression and correspondence analysis to screen the risk factors and the relationship between prevalence group and gene. The analysis showed: (1) 24 genes at significant risk of being detected in all the herds with infection prevalence >5%, including genes belonging to microbial surface components recognizing adhesive matrix molecules (MSCRAMMs), immune evasion and serine proteases; and (2) a significant correlation coefficient between the genes interacting with the host immune response and HP isolates against LP ones. These results support the hypothesis that virulence factors, in addition to cow management, could be related to strain contagiousness, offering new insights into vaccine development.

Effectiveness of Dry Cow Therapy Comprising Antibiotic Treatment, Internal Teat Sealant, and α-Tocopherol Against New Intramammary Infections in Cows

Abstract The aim of this study was to evaluate the preventive effectiveness of dry cow therapy based on antibiotic, internal teat sealant, and α-tocopherol administered separately or in various combinations at drying-off The study was performed on 322 uninfected quarters of 95 cows originating from three dairy herds. The new intramammary infection rates after calving were measured to evaluate the effectiveness. The quarters were divided into six groups differing in treatment, namely: control group (group C, n = 40) and five treatment groups. Treatment groups were arranged as follows: group A (antibiotic alone, n = 81), group AS (antibiotic + sealant, n = 40), group AST (antibiotic + sealant + α-tocopherol, n = 40), group T (α-tocopherol alone, n = 40), group S (sealant alone, n = 81). New infection rate amounted to 47.5% in group C. The treatment in group AST significantly prevented from the occurrence of new intramammary infections (12.5%, P &lt; 0.05), especially those caused by major pathogens. Antibiotic treatment alone (group A) did not prevent from new infections (34.6%, P &gt; 0.05), although the use of the sealant alone (group S) decreased the risk of new infection (24.7%, P &lt; 0.05). A decrease in new infection rate (25%, P &lt; 0.05) was also observed in AS group treated with the combination of the sealant and antibiotic. α -tocopherol supplementation alone (group T) had no overall effect on new infections (35%, P &gt; 0.05). Increased α-tocopherol level (P &lt; 0.05) was detected after calving in the quarters from cows that received α-tocopherol injections. In conclusion, the combination of antibiotic, internal teat sealant, and α-tocopherol used in dry cow therapy showed a significantly better preventive effect against new intramammary infections, than the therapeutics administered separately.

Characterization of Aerococcus viridans isolated from milk samples from cows with mastitis and manure samples.

Thirty-eight Aerococcus viridans isolates were obtained from milk from 478 cows with clinical mastitis in a farm during the periods between November 2011 and February 2012, and between December 2012 and March 2013. Additional isolates were obtained from processed manure (a mixture of composted manure, straw and hydrated lime) and bedding materials. The processed manure was later used to cover the floor of the stalls in barns as bedding materials. The temperatures recorded in the composted and processed manure were not as high as those generally observed during satisfactory composting. To reveal the association of A. viridans in manure-related products with intramammary infection in cows, isolates were characterized by their DNA fragment patterns as determined by pulsed-field gel electrophoresis (PFGE) and antimicrobial susceptibility testing. Isolates obtained from milk, processed manure and bedding materials had identical DNA fragment patterns. Antimicrobial susceptibilities were determined for 29 isolates from milk, processed manure and bedding materials. Of these, 26 (89.7%) were resistant to clindamycin, whereas virtually all the isolates were susceptible to 12 other antimicrobials including cefalosporins that have been used to treat bovine mastitis in Japan. In vitro, three A. viridans isolates from milk and an isolate from processed manure survived for 3 hr in Good’s buffer (pH 9) at high temperature (50°C). The results suggest that the processed manure and bedding materials in this farm were possible sources of A. viridans that caused infection in the cows with mastitis.

Effects of feed restriction and prolactin-release inhibition at drying-off on susceptibility to new intramammary infection in cows

A cow’s risk of acquiring a new intramammary infection during the dry period increases with milk production at drying-off. A method commonly used to reduce milk production is a drastic reduction in feed supply in the days that precede drying-off. Milk production can also be reduced by inhibiting the lactogenic signal driven by prolactin (PRL). This study aimed to compare the effects of these 2 drying-off procedures on milk production, metabolism, and susceptibility to intramammary infection in cows. A total of 21 Holstein cows in late lactation were assigned to 1 of 3 treatments based on milk yield, somatic cell count, and parity. The cows were fed a lactation diet until drying-off (control), only dry hay during the last 5 d before drying-off (DH), or the same diet as the control cows but with twice-daily i.m. injections of 4mg of quinagolide, a specific inhibitor of PRL release, from 5 d before drying-off until 13 d after (QN). On d 1 to 7 after the last milking, the cows were challenged by daily teat dipping in a solution containing Streptococcus agalactiae at 5×107 cfu/mL. Quinagolide induced a decrease in PRL concentration in blood on all the injection days. Blood PRL was also depressed in the hay-fed cows before drying-off. Both the QN and DH treatments induced a decrease in milk production, which at drying-off averaged 12.0, 10.0, and 21.7kg/d for the QN, DH, and control cows, respectively. The DH treatment decreased blood concentration of glucose and increased blood concentrations of β-hydroxybutyrate and nonesterified fatty acids before drying-off. Somatic cell count at drying-off was greater in the milk of the QN cows than in that of the control cows but after drying-off was greater in the mammary secretions of the control cows than in those of the QN cows. The number of S. agalactiae colonies found in mammary secretions on d 8 and 14 after the last milking was lower for the QN cows than for the control cows. The percentage of S. agalactiae-infected quarters was also lower in the QN cows than in the control cows and on d 14 averaged 17.2, 33.7, and 57.5% in the QN, DH, and control cows, respectively. No differences between the DH and control groups were observed for either bacterial count or infection rate. In conclusion, this experiment shows that PRL-release inhibition could be an alternative for reducing milk production and improving resistance to intramammary infection at drying-off.

Staphylococcus aureus shedding pattern throughout lactation in dairy cows with naturally occurring intramammary infection

To evaluate shedding patterns of Staphylococcus aureus, specifically the association between clonal relatedness and shedding patterns of S aureus for cows with naturally occurring S aureus intramammary infection. Longitudinal field study. Milk samples from 22 lactating cows (29 mammary glands) of varied numbers of lactations on 2 dairies. Procedures-Foremilk samples were collected weekly for 26 to 44 weeks during lactation from individual mammary glands. Milk samples were cultured bacteriologically with a 0.01-mL inoculum. Samples were considered culture positive for S aureus if ≥ 1 colony-forming units were obtained. Milk samples from known S aureus-positive mammary glands that were culture negative for S aureus or culture positive with a single colony of S aureus were cultured bacteriologically a second time with a 0.1-mL inoculum. Longitudinal shedding patterns of S aureus and the effect of strain type on ln(colony forming unit count) were examined. With the 0.01-mL inoculum, 914 of 1,070 (85%) samples were culture positive. After reculturing of negative samples with a 0.1-mL inoculum, 1,011 (95%) of the samples were culture positive. There was no significant difference in the detection of S aureus between genotypic clusters when either the 0.01- or 0.1-mL inoculum was used. There was no significant difference in the amount of shedding between mammary glands infected with isolates in genotypic cluster 1 or 2. No consistent shedding patterns were identified among or within cows. There was a significant difference in mammary gland linear score and test day (composite) linear score between mammary glands infected with isolates in genotypic clusters 1 and 2. S aureus was shed consistently in cows with naturally occurring intramammary infection in cows, and regardless of the pulsotype, variations in the amount of S aureus shedding had no significant effect on the ability to detect S aureus with a 0.1-mL inoculum.

Evaluation of a Petrifilm-based on-farm culture system for the detection of intramammary infections in cows with low somatic cell counts at dryoff

As an alternative to blanket dry-cow therapy, teat sealant-based selective dry cow therapy (SDCT) has the potential to greatly reduce the amount of antimicrobials used in dairy production. The main obstacle to SDCT is the necessity for a rapid and cost-effective method of identifying which cows have an infection at dry-off and would benefit from dry-cow antimicrobial therapy (DCT). Ideally, this method of diagnosis would be applicable to on-farm use and be simple to interpret. Because failing to treat an infected quarter at the end of lactation can have detrimental effects on the subsequent lactation, the diagnostic test should also have high sensitivity. Petrifilms are a ready-made culture media that provide results within 24 hours and are suitable for on-farm use. The presence of an indicator dye in the Petrifilm medium makes aerobic bacterial colonies appear as bright pink dots against a white background. Petrifilms have been validated as a diagnostic tool for the selective treatment of mastitis in lactating cows. The objective of this study was to determine the utility of a Petrifilm-based on-farm culture system for making treatment decisions for cows with low somatic cell counts (SCC) at dry-off.

Host response in bovine mastitis experimentally induced with Staphylococcus chromogenes

An experimental infection model was developed to study host response to intramammary infection in cows caused by Staphylococcus chromogenes. CNS intramammary infections have become very common in modern dairy herds, and they can remain persistent in the mammary gland. More information would be needed about the pathophysiology of CNS mastitis, and an experimental mastitis model is a means for this research. Six primiparous Holstein–Friesian cows were challenged with S. chromogenes 4 weeks after calving . One udder quarter of each cow was inoculated with 2.1 × 10 6 cfu of S. chromogenes. All cows became infected and clinical signs were mild. Milk production of the challenged quarter decreased on average by 16.3% during 7 days post-challenge. Cows eliminated bacteria in a few days, except for one cow which developed persistent mastitis. Milk indicators of inflammation, SCC and N-acetyl-β- d-glucosaminidase (NAGase) returned to normal within a week. Milk NAGase activity increased moderately, which reflects minor tissue damage in the udder. Concentrations of serum amyloid A (SAA) and milk amyloid A (MAA) were both elevated at 12 h PC. MAA was affected by the milking times, and was at its highest before the morning milking. In our experimental model, systemic acute phase protein response with SAA occurred as an on–off type reaction. In conclusion, this experimental model could be used to study host response in CNS mastitis caused by the main CNS species and also for comparison of the host response in a mild intramammary infection and in more severe mastitis models.

Evaluation of interleukin-2 treatment for prevention of intramammary infections in cows after calving

A low-dose treatment based on interleukin-2 (IL-2) was investigated for preventing mastitis in dairy cows. The treatment consisted of a single dose of IL-2 injected into the skin region drained by the supramammary lymph node 3-5 days after calving. The study included 45 cows (23 treated and 22 controls) from three commercial dairy herds. The results showed that the treatment had no side effects. The treatment with IL-2 induced the significant increase of several milk markers related to leukocyte and epithelial cell functions, i.e. SCC (somatic cell counts), serum amyloid A (SAA), lactoferrin and NAGase. The increased concentration of milk markers suggested also an activity of IL-2 on epithelial cells, resulting in a higher resistance to invading pathogens. Indeed, the increased efficiency of cells in the udder is supported by the higher frequency of healthy quarters observed in the treated group until day 17-19 after calving, in comparison with the control one.