Joanne Karzis

Sensitivity and specificity of a hand-held milk electrical conductivity meter compared to the California mastitis test for mastitis in dairy cattle

Screening tests for mastitis can play an important role in proactive mastitis control programs. The primary objective of this study was to compare the sensitivity and specificity of milk electrical conductivity (EC) to the California mastitis test (CMT) in commercial dairy cattle in South Africa using Bayesian methods without a perfect reference test. A total of 1848 quarter milk specimens were collected from 173 cows sampled during six sequential farm visits. Of these samples, 25.8% yielded pathogenic bacterial isolates. The most frequently isolated species were coagulase negative Staphylococci (n=346), Streptococcus agalactiae (n=54), and Staphylococcus aureus (n=42). The overall cow-level prevalence of mastitis was 54% based on the Bayesian latent class (BLC) analysis. The CMT was more accurate than EC for classification of cows having somatic cell counts >200,000/mL and for isolation of a bacterial pathogen. BLC analysis also suggested an overall benefit of CMT over EC but the statistical evidence was not strong (P=0.257). The Bayesian model estimated the sensitivity and specificity of EC (measured via resistance) at a cut-point of >25 mΩ/cm to be 89.9% and 86.8%, respectively. The CMT had a sensitivity and specificity of 94.5% and 77.7%, respectively, when evaluated at the weak positive cut-point. EC was useful for identifying milk specimens harbouring pathogens but was not able to differentiate among evaluated bacterial isolates. Screening tests can be used to improve udder health as part of a proactive management plan.

Comparing effects of freezing at -196 °C and -20 °C on the viability of mastitis pathogens

The aim of this study was to compare the effects of cryopreservation at approximately -196 °C in liquid nitrogen (N) and freezing at approximately -20 °C in a freezer, on the viability and survival of eight different mastitogenic bacteria inoculated in milk. Bacteria were frozen at approximately -20 °C in a freezer and cryopreserved at approximately -196 °C in liquid nitrogen. An effective preservation method was needed for follow-up samples from cows identified in the South African National Milk Recording Scheme (NMRS) with somatic cell counts above 250 000 cells/mL milk. The organisation responsible for sample collection of the NMRS milk samples also provides producers with liquid nitrogen for their semen flasks at the collection sites. This existing mode of storage and transport could therefore be utilised.Ten samples of each organism were thawed and cultured bi-weekly until week 18 for both temperature treatments. An additional sampling was performed at week 30 for samples frozen at approximately -20 °C. Freezing and cryopreservation did not impair subsequent isolation of Streptococcus dysgalactiae, Streptococcus uberis, Enterococcus faecalis, Staphylococcus aureus (STH) (phage type lytic group III) or Sta. aureus (STA) (phage typed, other than lytic group III). Survival was indicated by the isolation of bacteria from samples, and viability by the strength of growth of the bacteria isolated. The survival of Streptococcus agalactiae decreased after week 12 and Escherichia coli after week 16 of freezing, but both organisms survived under cryogenic preservation until week 18. Coagulase-negative staphylococci survived until week 18 for both freezing and cryogenic preservation.Both storage methods could thus contribute to the improvement of a pro-active approach towards udder health management in South African dairy herds.

Somatic cell count thresholds in composite and quarter milk samples as indicator of bovine intramammary infection status

The objective of the study was to establish an operational somatic cell count (SCC) threshold to predict the presence of intramammary infection (IMI) in composite milk samples and compare findings with those in quarter milk samples. South African dairy producers now preferred composite milk samples for herd udder health analysis because of increasing cow numbers, convenience of sampling and lower cost. A retrospective study was conducted on 345 461 composite and 89 638 quarter milk samples from South African herds. Variance estimates for the proportion of quarter samples testing positive were adjusted to account for the lack of their independence within individual cows. The IMI at SCC thresholds of 150 000 cells/mL and 200 000 cells/mL differed only by 3.26% in composite milk samples. Youden’s index indicated the optimum SCC thresholds for composite and quarter milk samples as 150 000 cells/mL and 200 000 cells/mL, respectively. At 150 000 cells/mL, sensitivity (95% confidence intervals [CI]) in composite milk samples was 65.3% (64.0%, 66.6%) and specificity was 66.8% (65.7%, 67.9%); and in quarter milk samples, sensitivity at 200 000 cells/mL was 70.8% (69.5%, 72.0%) and specificity was 63.6% (62.4%, 64.8%). The likelihood of infection for udders and quarters, respectively, was 1.034 and 1.327 at an SCC threshold of 150 000 cells/mL and 0.864 cells/mL and 1.177 cells/mL at 200 000 cells/mL. The area under the curve of the receiver operating characteristics graph was 0.7084 and 0.7277 for composite and quarter samples, respectively, indicating that the SCC test could be considered as a good indicator of IMI in both sample types.

A pathogen-specific approach towards udder health management in dairy herds: Using culture and somatic cell counts from routine herd investigations

A dedicated udder health diagnostic programme was developed and used over a 15-year period in South Africa to analyse milk samples based on microbiological and cytological patterns within various groups and for individual cows and udder quarters in dairy herds. These pathogen-specific analyses are utilised for pro-active improvement and management of udder health in South African commercial dairy herds. The programme acts as a monitoring tool and identifies management areas at risk and individual cows with udder disease and uses both quarter and composite milk samples. Intra-mammary infection (IMI) is a dynamic situation and depending on the time a milk sample is taken, false-negative results may be obtained. A new IMI and an infection that is curing may both have low somatic cell counts (SCCs), masking the true bacterial status. SCC in individual infected udder quarters may differ greatly depending on the causative bacterial species, its pathogenicity, the host immune status and the environmental factors involved. A pathogen-specific udder health approach was followed with repeated herd tests to take account of these udder health dynamics. The results of the herd IMI investigation are applied in practice to assist veterinarians, udder health consultants and managers to make informed and specific detailed decisions at both a herd and on an individual cow basis regarding udder health.

Host adapted intramammary infections in pregnant heifers which were co-housed and reared on fresh milk as calves.

BackgroundHeifers can calve down with intramammary infections (IMI) and udder damage. This will have a negative impact on their longevity, future milk yield and financial return. Co-housed pre-weaned calves that are fed fresh milk have the opportunity to suckle each other’s teats and may infect udders of fellow heifer calves with pathogens present in milk. The prevalence of IMI in pregnant heifers in South Africa (SA) which were co-housed and reared on fresh milk as calves, is not known. Quarter secretion samples from both pregnant heifers (n = 2065) and dry cows (n = 5365) were collected for microbiological analysis from eight SA dairy herds. All heifers tested in this study were co-housed pre-weaning and were fed fresh milk as calves.ResultsThe prevalence of coagulase negative staphylococci, Staphylococcus aureus, Streptococcus agalactiae, environmental streptococci, coliforms and samples with no bacterial growth in heifers was 26%, 0.9%, 0.08%, 1.4%, 0.4% and 66%, respectively. The overall prevalence ratio between heifers and cows for Staphylococcus aureus IMI was 0.76 (95% CI: 0.59, 0.98). Four of the individual herds had prevalence ratios of less than one (p < 0.05), one herd had a prevalence ratio of 3.15 (95% CI: 1.52, 6.32), and the remaining 3 herds had a prevalence ratio not significantly different from 1.0. Marginally significant differences were found between Staphylococcus aureus IMI in pregnant heifers compared to cows in their second and later lactations (p = 0.06, p = 0.05, respectively) but no significant differences between heifers and cows in their first lactation.ConclusionsThe presence of Streptococcus agalactiae IMI in heifers came as a surprise, especially as herd infection rates were low. The high prevalence ratio of Staphylococcus aureus between heifers and cows in one herd warrants further investigation due to the potential danger of udder damage in a young cow at the start of her productive life. The IMI in heifers with host adapted pathogens can also act as a source of new IMI for lactating dairy cows.

A cost-benefit model comparing the California Milk Cell Test and Milk Electrical Resistance Test

The indirect effects of mastitis treatment are often overlooked in cost-benefit analyses, but it may be beneficial for the dairy industry to consider them. The cost of mastitis treatment may increase when the duration of intra-mammary infections are prolonged due to misdiagnosis of host-adapted mastitis. Laboratory diagnosis of mastitis can be costly and time consuming, therefore cow-side tests such as the California Milk Cell Test (CMCT) and Milk Electrical Resistance (MER) need to be utilised to their full potential. The aim of this study was to determine the relative benefit of using these two tests separately and in parallel. This was done using a partial-budget analysis and a cost-benefit model to estimate the benefits and costs of each respective test and the parallel combination thereof. Quarter milk samples (n= 1860) were taken from eight different dairy herds in South Africa. Milk samples were evaluated by means of the CMCT, hand-held MER meter and cyto-microbiological laboratory analysis. After determining the most appropriate cut-off points for the two cow-side tests, the sensitivity and specificity of the CMCT (Se= 1.00, Sp= 0.66), MER (Se= 0.92, Sp= 0.62) and the tests done in parallel (Se= 1.00, Sp= 0.87) were calculated. The input data that were used for partial-budget analysis and in the cost-benefit model were based on South African figures at the time of the study, and on literature. The total estimated financial benefit of correct diagnosis of host-adapted mastitis per cow for the CMCT, MER and the tests done in parallel was R898.73, R518.70 and R1064.67 respectively. This involved taking the expected benefit of a correct test result per cow, the expected cost of an error per cow and the cost of the test into account. The CMCT was shown to be 11%more beneficial than the MER test, whilst using the tests in parallel was shown to be the most beneficial method for evaluating the mastitis-control programme. Therefore, it is recommended that the combined tests should be used strategically in practice to monitor udder health and promote a pro-active udder health approach when dealing with host-adapted pathogens.

Proactive udder health management in South Africa and monitoring of antibiotic resistance of Staphylococcus aureus in dairy herds from 2001 to 2010

Antibiotic resistance of strains of Staphylococcus aureus isolated from bovine milk is of concern internationally. The objective of this study was to investigate trends of resistance of S. aureus to antibiotics administered to dairy cows in 19 South African and one Zambian dairy herds (participating in the South African proactive udder health management programme) and to identify possible contributing factors. The resistance of S. aureus strains to eight commonly used antibiotics in South Africa from 2001 to 2010 was evaluated. Staphylococcus aureus isolates (n = 2532) were selected from cows with subclinical mastitis in 20 herds routinely sampled as part of the proactive udder health management programme. The isolates were selected from milk samples that had somatic cell counts more than 400 000 cells/mL and were tested for antibiotic resistance using a standard Kirby–Bauer test with published clinical breakpoints. The prevalence of antibiotic resistance was evaluated as a percentage of S. aureus isolates susceptible out of the total numbers for each antibiotic selected per year. Staphylococcus aureus showed a significant increase in percentage of susceptible isolates over time for all antibiotics tested except for ampicillin. The overall prevalence of mastitis did not change during the study period. However, the prevalence of mastitis caused by S. aureus (mostly subclinical cases) in the selected herds decreased numerically but not significantly. Reduction in the incidence of antibiotic resistance shown by S. aureus was presumed to be a result of the application of the proactive udder health management programme. The fact that the overall prevalence of mastitis was kept stable was possibly because of the influence of the management programme in conjunction with the return of infections caused by non-resistant strains.

Epidemiological and partial budget analysis for treatment of subclinical Staphylococcus aureus intramammary infections considering microbiological and cytological scenarios

An innovative method was investigated to aid in the elimination of Staphylococcus aureus (S. aureus) intramammary infections (IMI) from dairy herds. A stochastic model explore the economic benefit of three-day or eight-day treatment of subclinical IMI in all S. aureus infected cows or in only those with a somatic cell count (SCC) exceeding 200,000 cells/ml. An epidemiological model was developed to run parallel to the economic model that would predict the S. aureus IMI likely to persist, develop new infections and clinical mastitis. In the economic model a first algorithm was used to consider the low prevalence (LP) scenario and made use of S. aureus prevalence information provided by retrospective analysis of microbiological and cytological results in South Africa (2008-2012). The data used considered Staphylococcus aureus prevalence from [1.495; 1.595]95% to [6.72; 6.95]95% for SCC≤200,000 and SCC>200,000 cells/ml respectively. A second algorithm considered the high prevalence (HP) scenario to evaluate a simulated situation with a 5[U1] [R12] to 25% prevalence. Scenarios of low or high transmission ratio (TR) were included in the model according to the hygiene management on the farm. Probabilities and costs were calculated over 255days. The economic models predicted average cost indices for low S. aureus IMI and low TR to vary from -3179 ZAR (South African Rands) when subclinical cases with SCC higher than 200,000 cell/ml were treated for eight days, to -3663 ZAR when all subclinical S. aureus IMI were treated for three days. With a HP and high TR of S. aureus the average cost indices changed from -18,042 ZAR when none to -5433 ZAR per 255days when all S. aureus IMI were treated for eight days. The epidemiological model in this study predicted substantial benefit of treatment mainly in high TR scenarios. New IMI decreased up to77% in the three-day and up to 91% in the eight-day treatment scenarios. In the HP scenarios, persistent IMI were reduced by 94%. The number of clinical cases predicted with no treatment for subclinical infections was higher than the total number of clinical and subclinical cases in scenarios where cows were treated three or eight days. Initial prudent treatment of subclinical IMI resulted in less overall treatments and less new, persistent and clinical cases. Combined results of economic and epidemiological models indicated that the option that cost the least did not always have the best epidemiological outcome. Models may assist in optimising and balancing decisions relating to financial and IMI.

Validity of somatic cell count as indicator of pathogen-specific intramammary infections

The objective of this study was to determine whether somatic cell count (SCC) was an effective test, with a sensitivity exceeding 85%, to determine species-specific bacterial infections. In addition, the relation between the SCC and various udder pathogen groups was investigated. SCC thresholds of greater than 200 000 cells/mL were used in quarter and greater than 150 000 cells/mL in composite milk samples. A retrospective study was conducted on a data set for 89 635 quarter and 345 467 composite cow milk samples. Eleven SCC threshold values were used to evaluate the diagnostic efficacy for the following bacteria: Gram-positive major pathogens: Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus uberis; Gram-negative major pathogens: Escherichia coli, Klebsiella pneumonia and Serratia spp.; minor pathogens: coagulase-negative staphylococci, Micrococcus spp., Staphylococcus pseudintermedius, Streptococcus pyogenes, Enterococcus faecalis, Enterococcus canis, Trueperella pyogenes and other Enterobacteriaceae. Sensitivity and specificity were calculated taking the effect of clustering into account with quarter milk samples. Most samples yielding major Gram-positive pathogens (88.9% in quarter and 79.9% in composite samples) and minor pathogens (61.4% in quarter and 51.7% in composite samples) had SCC greater than 200 000 cells/mL. Sensitivity of the SCC test to detect major pathogens at an SCC threshold of greater than 200 000 cells/mL in quarter samples and greater than 150 000 cells/mL in composite milk samples was 88.2% and 84.2%, respectively, but specificity was low (57.7% and 52.8%, respectively).