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1.  Prevalence of subclinical mastitis in dairy farms in urban and peri-urban areas of Kampala, Uganda 
It is widely recognized that subclinical mastitis (SCM) is an extensive problem in the dairy industry worldwide. It is of particular concern in developing countries. The aim of this study was to establish the prevalence of SCM in dairy cattle in the urban and peri-urban areas of Kampala, Uganda and to gain information about pathogens and antibiotic resistance patterns. The study was conducted as a field study in 18 smallholder dairy farms in peri-urban Kampala, Uganda. All cows at the farms were physically examined, and cows with signs of clinical mastitis were excluded. Cows (n = 195) were tested with California Mastitis Test (CMT), and udder quarters with CMT score ≥3 (scale 1–5) were milk sampled for bacteriological analysis. To allow further sub-analysis of the results, the stage of lactation, parity, milk production, production type, udder hygiene, and cow breed were recorded. Results indicate that 86.2 % (n = 168) of the tested cows had SCM in one or more quarters. The most common bacteriological outcome was infection with coagulase-negative staphylococci (54.7 %), followed by negative growth (24.9 %) and streptococci (16.2 %); all of which (n = 34) were sensitive to penicillin. Of the tested staphylococci (n = 17), the majority (58.9 %) were positive for penicillinase production. Factors with significant impact on the prevalence of SCM at cow level were the stage of lactation, parity, and production type. The results suggest that the prevalence of SCM in Uganda is substantially higher than reported in previous studies and in other comparable developing countries. This implies that SCM deserves more attention and that improvement in dairy cow husbandry in terms of hygiene and management is necessary.
doi:10.1007/s11250-013-0455-7
PMCID: PMC3895220  PMID: 23955012
Udder health; Smallholder dairy farming; Holstein-Friesian cows; East Africa
2.  Comparison of α1-Antitrypsin, α1-Acid Glycoprotein, Fibrinogen and NOx as Indicator of Subclinical Mastitis in Riverine Buffalo (Bubalus bubalis) 
Mastitis set apart as clinical and sub clinical is a disease complex of dairy cattle, with sub clinical being the most important economically. Of late, laboratories showed interest in developing biochemical markers to diagnose sub clinical mastitis (SCM) in herds. Many workers reported noteworthy alternation of acute phase proteins (APPs) and nitric oxide, (measured as nitrate+nitrite = NOx) in milk due to intra-mammary inflammation. But, the literature on validation of these parameters as indicators of SCM, particularly in riverine milch buffalo (Bubalus bubalis) milk is inadequate. Hence, the present study focused on comparing several APPs viz. α1- anti trypsin, α1- acid glycoprotein, fibrinogen and NOx as indicators of SCM in buffalo milk. These components in milk were estimated using standardized analytical protocols. Somatic cell count (SCC) was done microscopically. Microbial culture was done on 5% ovine blood agar. Of the 776 buffaloes (3,096 quarters) sampled, only 347 buffaloes comprising 496 quarters were found positive for SCM i.e. milk culture showed growth in blood agar with SCC≥2×105 cells/ml of milk. The cultural examination revealed Gram positive bacteria as the most prevalent etiological agent. It was observed that α1- anti trypsin and NOx had a highly significant (p<0.01) increase in SCM milk, whereas, the increase of α1- acid glycoprotein in infected milk was significant (p<0.05). Fibrinogen was below detection level in both healthy and SCM milk. The percent sensitivity, specificity and accuracy, predictive values and likelihood ratios were calculated taking bacterial culture examination and SCC≥2×105 cells/ml of milk as the benchmark. Udder profile correlation coefficient was also used. Allowing for statistical and epidemiological analysis, it was concluded that α1- anti trypsin indicates SCM irrespective of etiology, whereas α1- acid glycoprotein better diagnosed SCM caused by gram positive bacteria. NOx did not prove to be a good indicator of SCM. It is recommended measuring both α1- anti trypsin and α1- acid glycoprotein in milk to diagnose SCM in buffalo irrespective of etiology.
doi:10.5713/ajas.2012.12261
PMCID: PMC4093252  PMID: 25049851
Acute Phase Proteins; Nitric Oxide; Subclinical Mastitis and Buffalo
3.  Udder health in beef cows and its association with calf growth 
Background
Studies outside the Nordic countries have indicated that subclinical mastitis (measured by milk somatic cell count or the California Mastitis Test), intramammary infections (IMI), or blind quarters in beef cows may have negative effects on beef calf growth. Knowledge on prevalence of such udder health problems in Swedish beef cows is scarce. Therefore, the main aim of this study was to investigate subclinical mastitis, IMI and udder conformation in a number of beef cow herds. Production of β-lactamase in staphylococci was also investigated. Associations between certain cow factors and subclinical mastitis and IMI, and associations between cow and calf factors and 200 day calf weaning weight were also studied. The herds were visited once within a month after calving and once at weaning. Udder examination and quarter milk sampling, for somatic cell count and bacteriology, were performed in 8 to 12 cows per herd and occasion.
Results
Approximately 50%, 40% and 10% of the cows had subclinical mastitis, IMI, and at least one blind quarter, respectively, but the prevalence varied markedly between herds. Intramammary infections (mainly due to staphylococci) were identified in 13-16% of the milk samples. Less than 5% of the staphylococcal isolates produced β-lactamase. Approximately 11% of the cows sampled twice had the same IMI (mostly Staphylococcus aureus) at both samplings. Cow factors of importance for subclinical mastitis and/or IMI were teat and udder shape, breed, parity, presence of blind quarters, and cow hygiene. No significant associations were found between udder health parameters studied and calf weaning weights.
Conclusions
Subclinical mastitis and IMI, but not blind quarters, were common in beef cows, but the prevalence varied markedly between herds. Most IMI were caused by staphylococci and more than 95% of those were sensitive to penicillin. Cows with large funnel-shaped teats or pendulous udder after calving, and cows with blind quarters were at risk of having subclinical mastitis and/or IMI. Poor hygiene was also a risk factor for udder health problems. No significant associations were found between udder health and calf weaning weight. More studies on risk factors are warranted to improve advisory services on awareness and prevention of mastitis in beef cows.
doi:10.1186/1751-0147-56-9
PMCID: PMC3922735  PMID: 24479386
Beef cows; Mastitis; Intra-mammary infections; Blind quarters; Risk factors; Calf weaning weight
4.  Effect of somatic cell count and mastitis pathogens on milk composition in Gyr cows 
Background
Gyr cows are well adapted to tropical conditions, resistant to some tropical diseases and have satisfactory milk production. However, Gyr dairy herds have a high prevalence of subclinical mastitis, which negatively affects their milk yield and composition. The objectives of this study were (i) to evaluate the effects of seasonality, mammary quarter location (rear x front), mastitis-causing pathogen species, and somatic cell count (SCC) on milk composition in Gyr cows with mammary quarters as the experimental units and (ii) to evaluate the effects of seasonality and somatic cell count (SCC) on milk composition in Gyr cows with cows as the experimental units. A total of 221 lactating Gyr cows from three commercial dairy farms were selected for this study. Individual foremilk quarter samples and composite milk samples were collected once a month over one year from all lactating cows for analysis of SCC, milk composition, and bacteriological culture.
Results
Subclinical mastitis reduced lactose, nonfat solids and total solids content, but no difference was found in the protein and fat content between infected and uninfected quarters. Seasonality influenced milk composition both in mammary quarters and composite milk samples. Nevertheless, there was no effect of mammary quarter position on milk composition. Mastitis-causing pathogens affected protein, lactose, nonfat solids, and total solids content, but not milk fat content. Somatic cell count levels affected milk composition in both mammary quarters and composite samples of milk.
Conclusions
Intramammary infections in Gyr cows alter milk composition; however, the degree of change depends on the mastitis-causing pathogen. Somatic cell count is negatively associated with reduced lactose and nonfat solids content in milk. Seasonality significantly affects milk composition, in which the concentration of lactose, fat, protein, nonfat solids and total solids differs between dry and wet seasons in Gyr cows.
doi:10.1186/1746-6148-9-67
PMCID: PMC3626830  PMID: 23566405
Somatic cell count; Gyr cows; Mastitis; Milk composition
5.  Characterization of coagulase negative staphylococci from cases of subclinical mastitis in dairy cattle in Kampala, Uganda 
Irish Veterinary Journal  2014;67(1):12.
Background
Coagulase negative staphylococci (CNS) are the most common pathogens leading to subclinical mastitis (SCM) in dairy cattle in Uganda. Coagulase negative staphylococci can vary between bacterial species in how they cause disease. The aim of the study was to characterize CNS, from cows with SCM in Uganda, at the species level.
Findings
Quarter milk samples (n = 166) were collected from 78 animals with SCM. Bacteriological analyses were carried out at Makerere University, Kampala, Uganda and at the National Veterinary Institute (SVA), Uppsala, Sweden. The most common pathogens found in milk samples from cows with SCM were CNS (31.7%). Two species of CNS were found, S. epidermidis (85%) and S. haemolyticus (15%). Of the CNS isolates, 16/20 (80%) were positive for β-lactamase production (β+).
Conclusions
In milk samples from cows with SCM caused by CNS, S. epidermidis was most prevalent, followed by S. haemolyticus.
doi:10.1186/2046-0481-67-12
PMCID: PMC4051667  PMID: 24917926
CNS; Staphylococcus epidermidis; Staphylococcus haemolyticus; MALDI-TOF
6.  Therapeutic Effect of Nisin Z on Subclinical Mastitis in Lactating Cows▿  
Bovine subclinical mastitis is an inflammation of the mammary gland caused by bacterial intramammary infection, accounting for large economic losses. Treatment of subclinical mastitis is not suggested for lactating cows due to the risk of milk contamination. The objectives of this study were to evaluate an antimicrobial peptide, nisin, in the treatment of subclinical mastitis in lactating cows. A total of 90 lactating Holstein cows with subclinical mastitis were randomly divided into nisin-treated (n = 46) and control (n = 44) groups. In the nisin-treated group, cows received an intramammary infusion of nisin at a dose of 2,500,000 IU once daily for 3 days while the control cows received no treatment. Milk samples were collected from the affected mammary quarters before treatment and 1 and 2 weeks after treatment for analyses of bacteria, somatic cells, and N-acetyl-β-d-glucosaminidase (NAGase). Results indicated that nisin therapy had bacteriological cure rates of 90.1% for Streptococcus agalactiae (10 of 11), 50% for Staphylococcus aureus (7 of 14), 58.8% for coagulase-negative staphylococci (7 of 17), and 65.2% for all cases (30 of 46). Meanwhile, only 15.9% (7 of 44) of untreated cows spontaneously recovered. NAGase activity in milk samples and the number of mammary quarters with a milk somatic cell count of ≥500,000/ml were significantly decreased after nisin treatment while no significant changes took place in the control group. Because of its therapeutic effects on bovine subclinical mastitis, as well as its safety in humans, nisin deserves further study to clarify its effects on mastitis caused by different pathogens.
doi:10.1128/AAC.00629-07
PMCID: PMC2043217  PMID: 17606675
7.  Occurrence of Clinical and Sub-Clinical Mastitis in Dairy Herds in the West Littoral Region in Uruguay 
Acta Veterinaria Scandinavica  2002;43(4):221-230.
Twenty-nine dairy farms were selected to determine the incidence of clinical mastitis, prevalence of sub-clinical mastitis and bacterial aetiology in the West Littoral Region of Uruguay. In samples taken by the owner and frozen at -20°C during a week the incidence rate of clinical mastitis was determined as 1.2 cases per 100 cow-months at risk. Staphylococcus aureus was the most common isolated pathogen in 37.5% of 40 milk samples from clinical cases obtained in 1 month. No bacteria grew in the 32.5% of the total samples.
A sub-sample including 1077 dairy cows from randomly selected farms was used to determine the prevalence of sub-clinical mastitis. These samples were taken on one visit to each farm. The prevalence was 52.4% on a cow basis and 26.7% on an udder quarter basis. In 55.1% of the quarters of the selected animals with more than 300 000 cells/ml there was no growth. The isolated pathogens from sub-clinical cases and their relative frequencies were: Staphylococcus aureus 62.8%, Streptococcus agalactiae 11.3%, Enterococcus sp. 8%, coagulase-negative staphylococci 7.4%, Streptococus uberis 6.4%, Streptococcus dysgalactiae 1.8%, Escherichia coli 1.5% and Staphylococcus hyicus coagulase-positive 0.6%.
doi:10.1186/1751-0147-43-221
PMCID: PMC1764198  PMID: 12831175
mastitis; Uruguay; cows; incidence; prevalence
8.  Quarter and cow risk factors associated with the occurrence of clinical mastitis in dairy cows in the united Kingdom 
Journal of dairy science  2009;92(6):2551-2561.
Quarter and cow risk factors associated with the development of clinical mastitis (CM) during lactation were investigated during a 12-mo longitudinal study on 8 commercial Holstein-Friesian dairy farms in the southwest of England. The individual risk factors studied on 1,677 cows included assessments of udder and leg hygiene, teat-end callosity, and hyperkeratosis; body condition score; and measurements of monthly milk quality and yield. Several outcome variables for CM were used for statistical analysis, which included use of generalized linear mixed models. Significant covariates associated with an increased risk of CM were increasing parity, decreasing month of lactation, cows with very dirty udders, and quarters with only very severe hyperkeratosis of the teat-end. Thin and moderate smooth teat-end callosity scores were not associated with an increased risk for CM. Cows that recorded a somatic cell count >199,000 cells/mL and a milk protein percentage <3.2 at the first milk recording after calving were significantly more likely to develop CM after the first 30 d of lactation. There was no association between cow body condition score and incidence of CM. Of the cases of CM available for culture, 171 (26.7%) were confirmed as being caused by Escherichia coli and 121 (18.9%) confirmed as being caused by Streptococcus uberis. Quarters with moderate and very severe hyperkeratosis of the teat-end were at significantly increased risk of clinical E. coli mastitis before the next visit. Quarters with very severe hyperkeratosis of the teat-end were significantly more likely to develop clinical Strep. uberis mastitis before the next visit. There were strong trends within the data to suggest an association between very dirty udders (an increased risk of clinical E. coli mastitis) and teat-ends with no callosity ring present (an increased risk of clinical Strep. uberis mastitis). These results highlight the importance of individual quarter- and cow-level risk factors in determining the risk of CM associated with environmental pathogens during lactation.
doi:10.3168/jds.2008-1369
PMCID: PMC2690977  PMID: 19447987
clinical mastitis; risk factor; hyperkeratosis; udder hygiene score
9.  Evaluation of Milk Trace Elements, Lactate Dehydrogenase, Alkaline Phosphatase and Aspartate Aminotransferase Activity of Subclinical Mastitis as and Indicator of Subclinical Mastitis in Riverine Buffalo (Bubalus bubalis) 
Mastitis is a highly morbid disease that requires detection at the subclinical stage. Tropical countries like India mainly depend on milch buffaloes for milk. The present study was conducted to investigate whether the trace minerals viz. copper (Cu), iron (Fe), zinc (Zn), cobalt (Co) and manganese (Mn) and enzyme activity of lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in riverine buffalo milk can be used as an indicator of subclinical mastitis (SCM) with the aim of developing suitable diagnostic kit for SCM. Trace elements and enzyme activity in milk were estimated with Atomic absorption Spectrophotometer, GBC 932 plus and biochemical methods, respectively. Somatic cell count (SCC) was done microscopically. The cultural examination revealed Gram positive bacteria as the most prevalent etiological agent. A statistically significant (p<0.01) increase in SCC, Fe, Zn, Co and LDH occurred in SCM milk containing gram positive bacterial agents only. ALP was found to be elevated in milk infected by both gram positive and negative bacteria. The percent sensitivity, specificity and accuracy, predictive values and likelihood ratios were calculated taking bacterial culture examination and SCC≥2×105 cells/ml of milk as the benchmark. Only ALP and Zn, the former being superior, were found to be suitable for diagnosis of SCM irrespective of etiological agents. LDH, Co and Fe can be introduced in the screening programs where Gram positive bacteria are omnipresent. It is recommended that both ALP and Zn be measured together in milk to diagnose buffalo SCM, irrespective of etiology.
doi:10.5713/ajas.2011.11426
PMCID: PMC4092961  PMID: 25049573
Riverine Buffalo; Somatic Cell Count; Subclinical Mastitis; Indicator; Trace Minerals; Enzyme Activity
10.  Impact of Intramammary Treatment on Gene Expression Profiles in Bovine Escherichia coli Mastitis 
PLoS ONE  2014;9(1):e85579.
Clinical mastitis caused by E. coli accounts for significant production losses and animal welfare concerns on dairy farms worldwide. The benefits of therapeutic intervention in mild to moderate cases are incompletely understood. We investigated the effect of intramammary treatment with cefapirin alone or in combination with prednisolone on gene expression profiles in experimentally-induced E. coli mastitis in six mid-lactating Holstein Friesian cows. Cows were challenged with E. coli in 3 quarters and received 4 doses of 300 mg cefapirin in one quarter and 4 doses of 300 mg cefapirin together with 20 mg prednisolone in another quarter. At 24 h (n = 3) or 48 h (n = 3) post-challenge, tissue samples from control and treated quarters were collected for microarray analysis. Gene expression analysis of challenged, un-treated quarters revealed an up-regulation of transcripts associated with immune response functions compared to un-challenged quarters. Both treatments resulted in down-regulation of these transcripts compared to challenged, un-treated quarters most prominently for genes representing Chemokine and TLR-signaling pathways. Gene expression of Lipopolysaccharide Binding Protein (LBP), CCL2 and CXCL2 were only significantly down-regulated in cefapirin-prednisolone-treated quarters compared to un-treated controls. Down-regulation of chemokines was further confirmed on the basis of protein levels in milk whey for CXCL1, CXCL2 and CXCL8 in both treatments with a greater decrease in cefapirin-prednisolone-treated quarters. The data reveal a significant effect of treatment on cell recruitment with a more pronounced effect in cefapirin-prednisolone treated quarters. Provided a rapid bacteriological clearance, combination therapy may prevent neutrophil-induced tissue damage and promote recovery of the gland.
doi:10.1371/journal.pone.0085579
PMCID: PMC3891811  PMID: 24454893
11.  Risk factors associated with bacteriological cure, new infection, and incidence of clinical mastitis after dry cow therapy with three different antibiotics 
Journal of Veterinary Science  2011;12(3):227-233.
Factors affecting bacteriological cure rates (BCR) and new intramammary infections (IMI) during the dry period as well as clinical mastitis (CM) during early lactation were investigated in 414 German Holstein dairy cows receiving dry cow therapy. Cows were treated with either benethamine benzylpenicillin (300,000 IU), penethamate hydriodide (100,000 IU), and framycetin sulphate (100 mg, n = 136), or cefquinome (150 mg, n = 135), or benzathine cloxacillin (1,280 mg, n = 143). Overall BCR, IMI, and CM at parturition were 86.4%, 20.7%, and 4.3%, respectively. The three antibiotic treatments differed only in BCR, with cloxacillin yielding better results than the others. Udder quarters from cows with > 4 lactations had a higher risk of IMI and CM at calving. Chronic changes in udder tissues were linked to a lower BCR and were associated with a higher risk of CM during early lactation. The risk of CM at calving was higher in udder quarters with unspecific or subclinical mastitis before drying off. In conclusion, with antibiotic dry cow therapy, age and health status of the udder appear to be major determinants of IMI and CM during the dry period and early lactation, while BCR was associated with the antibiotic type and udder tissue status.
doi:10.4142/jvs.2011.12.3.227
PMCID: PMC3165151  PMID: 21897095
dry cow treatment; risk factors; treatment success
12.  The Use of Induced Mammary Infections for Evaluating Dry Cow Treatment Products II. Trial of a Proposed Method to Compare Three Levels of Novobiocin 
Infections were induced at the end of lactation in all udder quarters of 19 cows by the infusion of 0.2 mL of a 10-4 dilution in milk of a six hour milk culture of Staphylococcus aureus, strain 305 (A.T.C.C No. 29740). Two right or two left udder quarters were infected at 15 days and the opposite two five days before the last milking of lactation. Following the last milking all four udder quarters of eight cows were treated with 400 mg novobiocin in 10 mL of 2% aluminum monosterate in peanut oil, gelled. All udder quarters of eight other cows were treated with 50 mg novobiocin in the same vehicle and the udder quarters of three cows were treated with the vehicle only.
At calving, eight of 32 quarters treated with 400 mg novobiocin were still infected, as were 18 of 32 treated with 50 mg of novobiocin and all those quarters treated with vehicle only. Results were identical from udder quarters infected 15 and five days before drying off.
No significant differences were found between quarters in milk yield on the last day of lactation, nor the length of the dry period. An increasing number of udder quarters were infected at calving with increase in lactation age of the cow, although the small number of cows would not allow a firm conclusion. A significant difference in results was found between front and hind udder quarters, only five of 32 front quarters were infected at calving as compared to 21 to 32 hind quarters.
The method proposed was found to give essentially the same results as those from a large field trial using the same antibiotic. It should therefore be useful in evaluation trials of new antibiotic products for dry cow treatment.
PMCID: PMC1320017  PMID: 548164
13.  Insights into udder health and intramammary antibiotic usage on Irish dairy farms during 2003-2010 
Irish Veterinary Journal  2012;65(1):7.
By international standards, Ireland is a relatively small dairy producer. However, the industry plays a critical role to the national economy, accounting for approximately 3% of national gross domestic product. This paper presents insights into udder health and intramammary antibiotic usage on Irish dairy farms during 2003-2010, based on data from several sources. Three data sources were used, including data on milk recording data, intramammary antibiotic sales and animal health assessment. The milk recording data included a single unadjusted herd-level somatic cell count (SCC) value for each herd at each milk recording, being the arithmetic mean of cow-level SCC of each cow at that recording, weighted by cow-level yield. These data were used to calculate the percentage of herds each month where the unadjusted herd SCC exceeded 200,000 and 400,000 cells/mL. Two logistic generalised estimating-equations (GEE) models were developed, the outcome variable being either the probability that the monthly SCC of a herd was greater than 400,000 cells/mL or less than or equal to 200,000 cells/mL. Spring herds had a lower probability of a high SCC (> 400,000 cells/mL) during February to October compared to non-Spring herds but a higher probability between November to January. The odds of a high SCC were greater in 2005, 2006, 2009 and 2010 but less in 2007 and 2008 compared to 2004. Smaller herds had higher odds of having a high SCC compared to larger herds. We present the number of intramammary tubes and the quantity of active substance (kg) sold annually in Ireland during 2003-2010. We infer an incidence of clinical mastitis of 54.0 cases per 100 cow-years at risk, assuming 4 tubes per treatment regime, one affected quarter per cow, tubes restricted to clinical cases only and 100% of treated cases considered new cases, based on data collected on sales of in-lactation intra-mammary antibiotics. With differing assumptions, this estimate varied between 25.8 and 77.0 cases per 100 cow-years at risk. Using data on sales of dry cow therapy intra-mammary antibiotics, we also infer that most Irish dairy farmers use blanket dry cow therapy. It is important that Ireland has an objective understanding of current levels of udder health, to facilitate benchmarking and improvement into the future. Udder health is a concern on a number of Irish dairy farms. High SCC results were present throughout the year, but more marked towards the start and end of each milking season. Animal Health Ireland recently commenced a major national programme, CellCheck, in collaboration with a broad range of stakeholders, to support national SCC improvement. In this paper, relevant European and national legislation is also reviewed.
doi:10.1186/2046-0481-65-7
PMCID: PMC3376034  PMID: 22455802
14.  Experimental intramammary inoculation with Mycoplasma bovis in vaccinated and unvaccinated cows: effect on milk production and milk quality. 
The effect of vaccination on milk production was evaluated in vaccinated and control cows experimentally challenged in two of four quarters with live Mycoplasma bovis. During the first three weeks after experimental challenge, six of eight unchallenged quarters on vaccinated cows and seven of eight unchallenged quarters on control cows became infected. Most of these quarters secreted normal milk, with negative California Mastitis Test scores and maintained normal milk production throughout most of the study (although some quarters on control cows remained infected). All challenged quarters became infected, had strong California Mastitis Test reactions, and had a drastic (greater than 85%) loss in milk production. Thereafter, four of eight challenged quarters on control cows remained infected, had mostly positive California Mastitis Test scores, produced mostly normal-appearing milk, and recovered some productive capabilities. By the end of the study no M. bovis could be recovered from challenged quarters on vaccinated cows and the milk appeared mostly normal. The California Mastitis Test scores on these quarters, however, remained elevated and milk production remained very low.
PMCID: PMC1255190  PMID: 3756674
15.  A method to estimate the somatic cell count of milk from a mastitic quarter using composite somatic cell count. 
The purpose of the study was to estimate the extent to which one quarter could be inflamed relative to the other three quarters of a cow, given that knowledge only of the composite milk somatic cell count (SCC) was available. An algebraic relationship, which incorporated the parameters of composite milk SCC, production loss associated with composite milk SSC, SCC of milk from an inflamed quarter, and SCC from three non-inflamed quarters, was used to derive hypothetical estimates of the SCC in one inflamed quarter. A simple case was considered in which one quarter was mastitic, the SCC in milk of the noninflamed quarters was equal, and there was no production compensation by the noninflamed quarters. Previously published estimates were used for production loss associated with composite milk SCC. For moderate composite milk SCC (300,000-500,000 cells/mL) and low-to-moderate SCC in milk of noninflamed quarters (100,000 cells/mL), the SCC of milk from one inflamed quarter was predicted to be very high, ranging from about 1.26 million (log10 SCC = 6.1) to 6.3 million (log10 SCC = 6.8) cells/mL, and compatible with signs of clinical mastitis. These results suggest that in order to screen cows with clinical mastitis in only one quarter, composite milk SCC should be considerably lower than values presented previously by other investigators.
PMCID: PMC1255627  PMID: 2306671
16.  Clinical mastitis in dairy cattle in Ontario: frequency of occurrence and bacteriological isolates. 
The objective of this study was to describe the frequency of occurrence of clinical mastitis in dairy herds in Ontario. The study group consisted of 65 dairy farms involved in a 2-year observational study, which included recording all clinical mastitis cases and milk sampling of quarters with clinical mastitis. Lactational incidence risks of 9.8% for abnormal milk only, 8.2% for abnormal milk with a hard or swollen udder, and 4.4% for abnormal milk plus systemic signs of illness related to mastitis were calculated for 2840 cows and heifers. Overall, 19.8% of cows experienced one or more cases of clinical mastitis during location. Teat injuries occurred in 2.1% of lactations. Standard bacteriology was performed on pretreatment milk samples from 834 cows with clinical mastitis. The bacteria isolated were Staphylococcus aureus (6.7%), Streptococcus agalactiae (0.7%), other Streptococcus spp. (14.1%), coliforms (17.2%), gram-positive bacilli (5.5%), Corynebacterium bovis (1.7%), and other Staphylococcus spp. (28.7%). There was no growth in 17.7% of samples, and 8.3% of samples were contaminated. Clinical mastitis is a common disease in dairy cows in Ontario; approximately 1 in 5 cow lactations have at lease one episode of clinical mastitis. There is, however, considerable variation in the incidence of clinical mastitis among farms. The majority of 1st cases of clinical mastitis occur early in lactation, and the risk of clinical mastitis increases with increasing parity. Environmental, contagious, and minor pathogens were all associated with cases of clinical mastitis.
Images
PMCID: PMC1539829  PMID: 9442950
17.  Factors affecting cure when treating bovine clinical mastitis with cephalosporin-based intramammary preparations 
Journal of dairy science  2009;92(5):1941-1953.
Data were collated for an independent scientific analysis from 2 international, multicenter studies that had compared the efficacy of 3 different cephalosporin-containing intramammary preparations in the treatment of clinical mastitis in dairy cattle [cefalexin (first generation) in combination with kanamycin; cefquinome (fourth generation); and cefoperazone (third generation)]. Quarters were assessed using standard bacteriological techniques before treatment and at 16 and 25 d posttreatment. Additional data were also available on individual cows and study farms, including parity, breed, and cow somatic cell count histories, herd bulk milk somatic cell counts, and farm management regimens. Sufficient data for analysis were available from a total of 491 cases on 192 farms in 3 countries (United Kingdom, France, and Germany) with up to 16 cases being recruited from any one farm. Clinical cases were of diverse etiology, representing both contagious and environmental pathogens. Univariable analysis demonstrated that quarters in the cefalexin + kanamycin and cefquinome treatment groups were not significantly different from each other, but were both significantly more likely to be pathogen free posttreatment than quarters in the cefoperazone group. Multivariable analysis was undertaken using conventional random effects models. Two models were built, with the first incorporating only information available to the practitioner at the time of treatment and the second including all information collected during the study. These models indicated that country, pretreatment rectal temperature (above-normal temperature associated with an increased chance of being pathogen free posttreatment), individual cow somatic cell count (increased somatic cell count associated with a decreased chance of being pathogen free posttreatment), and pathogen (Staphylococcus aureus isolation associated with a decreased chance of being pathogen free posttreatment) were useful predictors of pathogen free status; parity, yield, bulk milk somatic cell counts, and other farm management factors were not. The importance of country in the analysis demonstrates the need to generate local data when assessing treatment regimens. In addition, these results suggest that the factors important in predicting the outcome of treatment of clinical mastitis cases may be dissimilar to those reported to affect the likelihood of cure when treating subclinical intramammary infections.
doi:10.3168/jds.2008-1497
PMCID: PMC2675183  PMID: 19389951
clinical mastitis; treatment; risk factor; cephalosporin
18.  Udder pathogens and their resistance to antimicrobial agents in dairy cows in Estonia 
Background
The goal of this study was to estimate the distribution of udder pathogens and their antibiotic resistance in Estonia during the years 2007-2009.
Methods
The bacteriological findings reported in this study originate from quarter milk samples collected from cows on Estonian dairy farms that had clinical or subclinical mastitis. The samples were submitted by local veterinarians to the Estonian Veterinary and Food Laboratory during 2007-2009. Milk samples were examined by conventional bacteriology. In vitro antimicrobial susceptibility testing was performed with the disc diffusion test. Logistic regression with a random herd effect to control for clustering was used for statistical analysis.
Results
During the study period, 3058 clinical mastitis samples from 190 farms and 5146 subclinical mastitis samples from 274 farms were investigated. Positive results were found in 57% of the samples (4680 out of 8204), and the proportion did not differ according to year (p > 0.05). The proportion of bacteriologically negative samples was 22.3% and that of mixed growth was 20.6%. Streptococcus uberis (Str. uberis) was the bacterium isolated most frequently (18.4%) from cases of clinical mastitis, followed by Escherichia coli (E. coli) (15.9%) and Streptococcus agalactiae (Str. agalactiae) (11.9%). The bacteria that caused subclinical mastitis were mainly Staphylococcus aureus (S. aureus) (20%) and coagulase-negative staphylococci (CNS) (15.4%). The probability of isolating S. aureus from milk samples was significantly higher on farms that had fewer than 30 cows, when compared with farms that had more than 100 cows (p < 0.005). A significantly higher risk of Str. agalactiae infection was found on farms with more than 600 cows (p = 0.034) compared with smaller farms. The proportion of S. aureus and CNS isolates that were resistant to penicillin was 61.4% and 38.5%, respectively. Among the E. coli isolates, ampicillin, streptomycin and tetracycline resistance were observed in 24.3%, 15.6% and 13.5%, respectively.
Conclusions
This study showed that the main pathogens associated with clinical mastitis were Str. uberis and E. coli. Subclinical mastitis was caused mainly by S. aureus and CNS. The number of S. aureus and Str. agalactiae isolates depended on herd size. Antimicrobial resistance was highly prevalent, especially penicillin resistance in S. aureus and CNS.
doi:10.1186/1751-0147-53-4
PMCID: PMC3041692  PMID: 21299911
19.  Frequency Tuning of the Cervical Vestibular-Evoked Myogenic Potential (cVEMP) Recorded from Multiple Sites along the Sternocleidomastoid Muscle in Normal Human Subjects 
Frequency tuning of tone burst-evoked myogenic potentials recorded from the sternocleidomastoid muscle (cervical VEMP or cVEMP) is used clinically to assess vestibular function. Understanding the characteristics of cVEMP is important for improving the specificity of cVEMP testing in diagnosing vestibular deficits. In the present study, we analyzed the frequency tuning properties of the cVEMPs by constructing detailed tuning curves and examining their morphology and dependence on SCM tonic level, sound intensity, and recording site along the SCM. Here we report two main findings. First, by employing nine tone frequencies between 125 and 4,000 Hz, some tuning curves exhibited two distinct peaks, which cannot be modeled by a single mass spring system as previously suggested. Instead, the observed tuning is better modeled as linear summation of two mass spring systems, with resonance frequencies at ~300 and ~1,000 Hz. Peak frequency of cVEMP tuning curves was not affected by SCM tonic level, sound intensity, and location of recording site on the SCM. However, sharpness of cVEMP tuning was increased at lower sound intensities. Second, polarity of cVEMP responses recorded from the lower quarter of the SCM was reversed as compared to that at the two upper sites. While more studies are needed, these results suggest that cVEMP tuning is mediated through multiple generators with different resonance frequencies. Future studies are needed to explore implications of these results on development of selective VEMP tests and determine the nature of polarity inversion at the lower quarter of SCM.
doi:10.1007/s10162-012-0360-1
PMCID: PMC3540270  PMID: 23183876
sound activation of vestibular system; canal; otolith; VCR
20.  A cohort study of coagulase negative staphylococcal mastitis in selected dairy herds in Prince Edward Island. 
The epidemiology and importance of coagulase negative staphylococcal (CNS) mastitis in Prince Edward Island had not been documented. To investigate this, a cohort of 84 cows at seven farms were quarter sampled eight times over a lactation, commencing with samples taken prior to drying off in the previous lactation. Thirteen species of CNS were isolated. The quarter prevalence of CNS mastitis varied from 4.8% to 6.4% in the first five months of lactation and increased to 14.2 to 16.6% in the last four months of lactation. The geometric mean somatic cell counts (SCC) for quarters infected with CNS and uninfected quarters were 90 x 10(3) and 64 x 10(3) respectively (difference significant at p > 0.005). The two month new infection risk of CNS was 9.0% while the two month elimination risk was 74.4%. Infection with CNS did not alter the risk of subsequent infection with Staphylococcus aureus. The results from this project support the classification of CNS as a minor pathogen in mastitis control programs.
PMCID: PMC1263556  PMID: 1477796
21.  Molecular Epidemiology of Streptococcus uberis Isolates from Dairy Cows with Mastitis 
Journal of Clinical Microbiology  2001;39(4):1460-1466.
Pulsed-field gel electrophoresis and antimicrobial sensitivity testing were used as tools to investigate the epidemiology of Streptococcus uberis mastitis in dairy cows. A total of 62 different strains were found among 138 isolates from the four herds investigated, and between 10 and 26 different strains were found in each herd. There was no strain common to all four herds. Identical strains of S. uberis were detected from different quarters of individual cows and from cows within the same herd, suggesting that transmission from quarter to quarter and cow to cow had occurred. Despite the great variation in S. uberis strains, persistent infection with the same strain within a lactation was observed in most cows. Predominant strains were present in two herds. Preliminary investigations could not clarify why these particular strains might predominate, but in one herd there was a significant difference between the prevalence of clinical mastitis in quarters infected with the predominant strain and that in quarters infected with other strains, suggesting the greater virulence of the predominant strain. The wide variety of S. uberis strains found is consistent with an environmental source of S. uberis. However, evidence of direct transmission, the persistence of infection, and the predominance of particular strains in some herds indicate that S. uberis infections are epidemiologically complex and that the relative importance of these factors in the occurrence of mastitis may differ between herds.
doi:10.1128/JCM.39.4.1460-1466.2001
PMCID: PMC87955  PMID: 11283072
22.  Posology and Field Efficacy Study with Novobiocin for Intramammary Infusion in Nonlactating Dairy Cows 
Four dose levels of novobiocin (50, 200, 400, 600 mg) were compared with no drug for the intramammary treatment of Staphylococcus aureus, Streptococcus agalactiae and other streptococcal infections present in the udder of dairy cows at the initiation of the dry period. Treatment success was evaluated by comparing the microbiological status of duplicate pretreatment quarter milk samples collected at drying off with the microbiological status of duplicate quarter milk samples collected four to ten days postcalving. Infection status of 1318 cows in 75 herds in five geographic locations was determined. Treatment effects on infected cows were evaluated by least squares analysis of variance with treatment, herd, lactation number, days dry and milk production at drying off considered as variables. The dose of 400 mg novobiocin per quarter was demonstrated to be significantly more effective (P < 0.05) than no drug and significantly better than (P < 0.05) or equal to the other doses for curing infections caused by S. aureus, S. agalactiae and other streptococci. A significant reduction (P < 0.05) in the overall rate of new udder infections acquired during the dry period was observed in cows treated with ≥ 200 mg novobiocin at drying off. The data supported the conclusion that the cow rather than the quarter is the appropriate experimental unit in the evaluation of intramammary mastitis treatments. Herd and lactation number were the most significant variables affecting cures.
PMCID: PMC1320019  PMID: 548166
23.  Effect of mastitis on raw milk compositional quality 
Journal of Veterinary Science  2007;8(3):237-242.
In this study, we investigated the impact of mastitis infection on the quality of milk composition in small-scale dairy bovine herds. The purpose of this study was to find a milk quality somatic cell count (SCC) standard that could be used as an integral component of a control program. In all, 396 quarter milk samples from lactating cross-bred cows (Holstein & Zebu) were analyzed; 56% of these quarters were experiencing intramammary infection, with an overall mean SCC of 5.46 × 105 ± 2.30 × 104cells/ml. Infected quarters had significantly (p < 0.05) higher mean SCC levels (6.19 × 105± 4.40 × 104cells/ml) compared to healthy quarters (2.65 × 105 ± 2.40 × 104cells/ml). In high SCC milk and infected quarters, the concentrations of non-casein fractions, sodium, chloride, and free fatty acid were higher (p < 0.05), while the casein content, lactose, casein-to-total protein, potassium, and calcium were lower (p < 0.05) compared to normal quarters. These findings suggest a mean SCC threshold limit of 5.46 × 105 cells/ml for the region. It was concluded that the results could be used to propose a milk quality SCC standard that can be used as an integral component of a control program.
doi:10.4142/jvs.2007.8.3.237
PMCID: PMC2868129  PMID: 17679769
compositional quality; raw milk; small scale farms; somatic cell count; subclinical mastitis
24.  A study of bovine mastitis, milking procedures and management practices on 25 Estonian dairy herds 
Background
Mastitis prevalence, milking procedures and management practices were investigated in 25 big dairy herds supplying milk to an Estonian dairy company. The aim of the study was to provide information for the company to be used in their new udder health improvement program to be set up after the completion of this study.
Methods
Quarter milk samples were collected from 3,166 cows for bacterial analysis and SCC (somatic cell counting). During the farm visit the veterinarian filled in a questionnaire about milking procedures and management practices with the help of farm managers. If the milk SCC of a cow or of a quarter exceeded 200,000/ml, the cow was defined as having mastitis.
Results
The percentage of cows having inflammation in one or more quarters measured by SCC (200,000/ml) was 52.7%. Corynebacterium bovis, Staphylococcus aureus and coagulase negative staphylococci were the most common bacterial isolates. Women as farm owners, and participating in the milking, were associated with lower mastitis prevalence on the farm. Peat bedding was associated with higher mastitis prevalence.
Conclusion
We demonstrated relatively high mastitis prevalence in this study. Contagious bacteria (eg. S. aureus, C. bovis, S. agalactiae and coagulase negative staphylococci) caused most of the infections. These infections are usually spread from cow to cow at milking if the milking hygiene is not good enough. The mastitis situation could be improved by improving milking procedures and hygiene.
doi:10.1186/1751-0147-48-22
PMCID: PMC1664578  PMID: 17118211
25.  Changes in thermal nociceptive responses in dairy cows following experimentally induced Escherichia coli mastitis 
Background
Mastitis is a high incidence disease in dairy cows. The acute stage is considered painful and inflammation can lead to hyperalgesia and thereby contribute to decreased welfare. The aim of this study was to examine changes in nociceptive responses toward cutaneous nociceptive laser stimulation (NLS) in dairy cows with experimentally induced Escherichia coli mastitis, and correlate behavioral changes in nociceptive responses to clinical and paraclinical variables.
Methods
Seven Danish Holstein-Friesian cows were kept in tie-stalls, where the E. coli associated mastitis was induced and laser stimulations were conducted. Measurements of rectal temperature, somatic cell counts, white blood cell counts and E. coli counts were conducted. Furthermore, scores were given for anorexia, local udder inflammation and milk appearance to quantify the local and systemic disease response. In order to quantify the nociceptive threshold, behavioral responses toward cutaneous NLS applied to six skin areas at the tarsus/metatarsus and udder hind quarters were registered at evening milking on day 0 (control) and days 1, 2, 3, 6 and 10 after experimental induction of mastitis.
Results
All clinical and paraclinical variables were affected by the induced mastitis. All cows were clinically ill on days 1 and 2. The cows responded behaviorally toward the NLS. For hind leg stimulation, the proportion of cows responding by stepping was higher on day 0 than days 3 and 6, and the frequency of leg movements after laser stimulation tended to decrease on day 1 compared to the other days. After udder stimulation, the proportion of cows responding by stepping was higher on day 1 than on all other days of testing. Significant correlations between the clinical and paraclinical variables of disease and the behavioral responses toward nociceptive stimulation were found.
Conclusions
Changes in behavioral responses coincide with peaks in local and systemic signs of E. coli mastitis. During the acute stage of E. coli mastitis nociceptive thermal stimulation on hind leg and mammary glands results in decreased behavioral responses toward nociceptive stimulation, which might be interpreted as hypoalgesia.
doi:10.1186/1751-0147-53-32
PMCID: PMC3114774  PMID: 21592395

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