Composite Milk Samples Research Articles

The effect of a commercial probiotic product on the milk quality of dairy cows

Probiotics intended to improve plant health and productivity of pastures grazed by dairy cow are becoming commercially available in Australia. Great Land (GL; Terragen Biotech Pty Ltd., Coolum Beach, QLD, Australia) is one such biologic soil conditioner and spray-on probiotic with a label claim of "acting to improve plant health and productivity." The objective of this study was to quantify the effect of GL on the milk quality of cows grazing pasture top-dressed with GL. Lactating dairy cows of mixed age and breed (primarily Holstein-Friesian), in their second lactation or greater, and at least 80 d in milk were enrolled and randomly allocated into 1 of 2 study groups: a treatment cow group (n = 98; cows grazed pasture that was top-dressed with GL according to the product label) and a control cow group (n = 114; cows grazed untreated pasture). As required, both groups were supplemented at the same rate with a mixed ration during the grazing period. Composite milk samples were collected weekly from each cow during the study and analyzed to determine milk components. Milk volumes were recorded at each milking using the herd management software of the study farm. Mean differences in the milk component variables were compared using mixed-effects linear regression models. After controlling for the effect of days in milk, cow lactation, and time since a cow entered the study, the treatment cows produced an average of 1.21 L/cow per day more milk (95% confidence interval: 0.34-2.08 L/cow per day) and more milk protein (0.03 kg/d; 95% confidence interval: 0.01-0.05 kg/d) than the control cows. Pasture cover and pasture consumption did not differ between the GL-treated and the untreated study paddocks grazed by the treatment or control cows. A limited amount of published data have examined the effect of probiotic pasture treatment on the milk quality of dairy cows. This study suggests that application of such products may be beneficial. The mechanisms associated with this type of outcome remain to be investigated.

Isolation and characterization of E. coli strains causing intramammary infections from dairy animals and wild birds

ABSTRACTThe study was conducted to estimate the prevalence of Escherichia coli (E. coli) in sub-clinically mastitic (SCM) animals, and in wild and migratory birds which may act as reservoir disseminating such pathogen. Farm hygiene, management and milking procedures were listed through a questionnaire. Thirty lactating cows and 15 lactating buffaloes from five small-scale dairy farms were randomly selected and screened for subclinical mastitis (SCM) using California Mastitis Test (CMT) and somatic cell count (SCC). In addition, 80 teat skin swabs, 5 drinking water samples and 38 wild and migratory bird faecal matter were also collected. All samples were processed for E. coli isolation by culturing on Levine’s Eosin Methylene Blue (L-EMB) agar, followed by purification and biochemical identification. Positive samples were subjected to molecular identification and serotyping. In addition, the presence of extended-spectrum beta-lactamase (ESBL) and carbapenemase-producing E. coli have been reported by antimicrobial sensitivity testing. Escherichia coli were isolated from 7.7%, 50% and 50% of the positive CMT cows’ quarters, cows’ composite and buffaloes’ composite milk samples, respectively. In addition, 14% of cows’ teats, 20% of water samples, 70% of faecal matter from wild bird, and 33.3% of faecal matter from migratory waterfowls were carrying E. coli. Serotyping, antibiotic-resistant pattern and phylogenetic analysis have pointed the bearable implication of milking hygiene and wild birds in disseminating E. coli strains causing intramammary infections.

Short communication: Validation of a test-day milk test for β-hydroxybutyrate for identifying cows with hyperketonemia

Subclinical ketosis is a common metabolic disorder affecting dairy cattle that results in a greater risk for the development of subsequent metabolic and infectious disease. Canwest Dairy Herd Improvement (DHI; Guelph, ON, Canada) has begun to use an infrared test (MilkoScan FT600, Foss Analytical A/S, Hillerød, Denmark) applied to metered composite milk samples to detect β-hydroxybutyrate (BHB) levels as a herd surveillance test for hyperketonemia. However, the test has not been compared with the gold standard, serum BHB as determined in a reference diagnostic laboratory. The objective of this cross-sectional diagnostic accuracy study was to validate the DHI milk BHB test to identify cows with hyperketonemia as determined by quantification of BHB in serum. A total of 316 cows from 17 dairy herds in southwestern Ontario had a milk and blood sample taken. Milk was collected at a routine DHI test, and blood from the same cow was sampled within 24 h of the milk test. The BHB concentration in milk was determined using the DHI milk BHB test, and serum was sent to the Animal Health Laboratory at the University of Guelph (Guelph, ON, Canada). A nonparametric receiver operating characteristic curve was generated to compare DHI milk BHB concentrations with serum BHB concentrations. Overall, a total of 34 cows (11%) had a level of serum BHB ≥1.2 mmol/L. The concentration of DHI milk BHB was moderately correlated with the concentration of serum BHB, yielding a coefficient of determination value of 0.61. The optimal cut point for determining hyperketonemia (≥1.2 mmol/L) on the DHI milk BHB test was ≥0.14 mmol/L, yielding a sensitivity of 81% and specificity of 92%. The performance of the DHI milk BHB test varied depending on the days in milk (DIM) of the cows tested, with a higher specificity being found in cows that were ≤25 DIM compared with cows tested >25 DIM. If the herd-level prevalence of hyperketonemia was ≥14%, the DHI milk BHB test had an improved sensitivity when compared with a herd-level prevalence of <14%. This study demonstrates that the DHI milk BHB test is a reliable measure for evaluating hyperketonemia using routine DHI milk samples and could be used as a herd-level monitoring tool for ketosis when evaluating nutritional management or preventative medicine strategies.

Development of Simplex-PCR assays for Accurate Identification of Nine Staphylococcal Species at Genus and Species Levels

Objective: Species identification of Staphylococcus is the prerequisite for precise assessment of&#x0D; microbial dynamics including their transmission and pathogenic significance in&#x0D; the dairy herd environment. The present study aimed to develop simplex PCR&#x0D; assays for rapid and specific identification of nine different Staphylococcus species. &#x0D; &#x0D; Methods: Specific&#x0D; primers targeting sodA gene for S. aureus, S. chromogenes, S. hominis,&#x0D; S. haemolyticus, S. hyicus; gap gene for S. sciuri, S. auricularis, S. simulans;&#x0D; and rdr gene for S. epidermidis were designed. The PCR assays were evaluated against&#x0D; 28 ATCC reference strains and 209 composite milk samples. Partial 16S rRNA sequencing was performed to&#x0D; reconfirm the results. &#x0D; &#x0D; Results: The&#x0D; PCR assays allowed species level identification of 348 staphylococcal field&#x0D; isolates recovered from 209 milk samples. The identification pattern was S. aureus (n=101), S. chromogenes (n=89), S.&#x0D; epidermidis (n=57), S. sciuri&#x0D; (n=43), S. haemolyticus (n=34), S. hyicus (n=13), S. hominis (n=5), S.&#x0D; auricularis (n=3) and S. simulans (n=3). The PCR based species&#x0D; identification was in 100% concordance with the partial 16S rRNA gene sequencing approach.&#x0D; &#x0D; Conclusion: The simplex PCR assays can be used as a precise tool&#x0D; for routine identification of Staphylococcus&#x0D; species from bovine milk as Staphylococcus&#x0D; species including coagulase-negative staphylococci is recognized a major cause&#x0D; of bovine mastitis in different parts of the world including India. J Microbiol Infect Dis 2018; 8(3):120-127

Diagnostic accuracy of Somaticell, California Mastitis Test, and microbiological examination of composite milk to detect Streptococcus agalactiae intramammary infections

The objectives of this study were to estimate the accuracy of Somaticell (Idexx Laboratories Inc., Westbrook, ME), California Mastitis Test (CMT), and microbiological examination of composite milk (MEC) to diagnose Streptococcus agalactiae intramammary infections (IMI), and to assess the agreement between Somaticell and CMT to detect these infections. A secondary objective was to estimate quarter- and cow-level prevalence of S. agalactiae IMI in the herds included in the study. Seven farms were included in the study. The CMT was performed and aseptic milk samples were collected from all quarters of all lactating cows. Composite milk samples were produced in the laboratory by mixing milk from all quarters of each sampled cow. The Somaticell test was performed on a subset of S. agalactiae-positive (n = 167) and S. agalactiae-negative (n = 152) quarter milk samples. Microbiological examination of quarter milk samples (MEQ) was considered the reference test for diagnosing S. agalactiae IMI. The accuracy of all tests at various thresholds was estimated using Bayesian latent class models. Apparent prevalence of S. agalactiae IMI was 15.8% (n = 184/1,164) at the quarter level (based on MEQ) and 28.5% (n = 83/291) at the cow level (based on MEC). True prevalence, as determined by Bayesian models, was 13.0% [95% credible interval (CR): 6.4-24.4%] at the quarter level, and 25.6% (95% CR: 15.3-39.5%) at the cow level. At the cow level (n = 285), sensitivity and specificity of MEC were 95.6 and 99.5%, respectively. The accuracy of Somaticell (n = 319 quarters) to identify S. agalactiae-infected quarters was 75.4, 86.4, 88.9, 89.4, and 91.0% at thresholds of 98,000, 147,000, 205,000, 244,000, and 282,000 cells/mL, respectively. The accuracy of CMT was 87.6, 90.7, 90.8, and 87.4% at thresholds of trace, 1, 2, and 3, respectively. The areas under the receiver operating characteristic curve for Somaticell and CMT were 94.5% (95% confidence interval: 91.8-97.2%) and 92.0% (88.6-95.4%), respectively. At the tested thresholds, the sensitivity of Somaticell ranged from 94.9 to 99.5% to detect S. agalactiae IMI, and specificity ranged from 48.1 to 87.1%. The sensitivity of Somaticell at the lowest threshold (69,000 cells/mL; sensitivity = 99.9%; 95% CR: 98.2-100%) was higher than that of CMT at any tested threshold. Results of this study could be used at the farm level to reduce the use of antimicrobials and reach specific goals in S. agalactiae eradication programs.

Subclinical intramammary infection does not affect bovine milk ethanol stability

The present study hypothesized that intramammary infection (IMI) might reduce milk ethanol stability (MES), mainly when IMI is caused by major pathogens. Thus, this study evaluated the effect of IMI on bovine MES using a natural exposure experimental design. Ninety-four lactating cows from five dairy herds were selected once they were determined to have an IMI, based on milk bacteriological culturing with positive isolation and somatic cell count (SCC) &gt; 200×103 cells/mL in two out of three composite milk samples collected during three consecutive weeks. After selection, cows were sampled a second time (within two weeks) for evaluation at mammary quarter level (n = 326): milk yield (kg/quarter/day), MES, composition (fat, protein, lactose, casein, total solids and solids-non-fat), and bacteriologic culture. The effect of subclinical mastitis on MES was tested by two models: 1) comparison of healthy vs. infected quarters; and 2) comparison of contralateral mammary quarter within cow. The only milk composition variable associated with MES was lactose (r = 0.18; P &lt; 0.01). Subclinical IMI did not affect MES when the comparison was performed using both models (1 and 2). Likewise, MES did not change when infected quarters were sorted into two groups of pathogens (major, minor and infrequent; and contagious, environmental, minor and infrequent) and compared with healthy mammary quarters. Considering the results of both models, subclinical IMI did not affect MES of dairy cows.

Addition of Curcumin in Dairy Sheep Diet in the Control of Subclinical Mastitis

Background: Curcumin, a polyphenol derived from rhizome of Curcuma longa, has been successfully used in animals and humans due to its antibacterial and anti-inflammatory properties. Recently, a study demonstrated that curcumin ointment reduced pain, mammary tension and erythema in humans with mastitis 72 h after the treatment. Mastitis in sheep is an important problem due to restrictions associated to the use of antibiotics, affecting milk production and becoming a public health issue. Thus, the aim of this study was to verify whether the addition of curcumin in the diet of six female sheep with subclinical mastitis could have curative efficacy and benefits to animal health.Cases: Six Lacaune sheep with subclinical mastitis that persisted for more than thirty days after the parturition were used. The animals received feed containing the curcumin (60 mg/animal/day) during 10 days. On days 0 and 10 of the experiment, total blood was collected to perform the hemogram and serum biochemistry (triglycerides, cholesterol, glucose, total protein (TP), albumin, globulin, aspartate aminotransferase (AST) and urea), as well as individual milk samples for centesimal composition, somatic cell count (SCC) and total bacterial count (TBC). Total leukocytes and erythrocytes, as well as hemoglobin content, were obtained using the hematological counter CELM-CC-550. Blood smears were stained by theRomanowsky method and used for leukocytes differentiation. After ten days of treatment, five sheep were negative in the California Mastitis Test (CMT). Total erythrocytes increased, while total leukocytes decreased after the treatment. Seric levels of AST, triglycerides, cholesterol and globulins decreased, while seric levels of albumin and urea increased after tendays of treatment. No differences were observed regarding milk production and composition, as well as for SCC and TBC on days 0 and 10 post treatment.Discussion: The use curcumin to prevent or to delay the occurrence of subclinical mastitis in ewes was first tested. In an experiment of lipopolysaccharide-induced mastitis in rats, researchers identified receptors type Toll 4 which are related with the induction of the synthesis of pro-inflammatory cytokines, as tumor necrosis factor alpha, interleukin-6 and 1β, molecules involved in the migration of leukocytes to the site of inflammation. According to literature, curcumin was able to reduce theactivity of myeloperoxidase in the mammary gland, an enzyme used as indicator of neutrophil infiltration, demonstrating the potent anti-inflammatory effect of curcumin. This effect can be associated to the release of interleukin-8, a cytokine that promotes the migration of neutrophils, lymphocytes and eosinophils into the site of infection, i.e., the mammary gland. It is important to highlight that neutrophils, due to its capacity to release inflammatory mediators and to delay apoptosis, has the potential to maintain a permanent inflammatory process, effects that were prevented or reduced by curcumin, specially the apoptotic and cytotoxic effects. It is important to emphasize the direct effect of curcumin on bacteria present in the mammary gland, since it exerts potent bactericidal effect. In summary, the use of curcumin in sheep with subclinical mastitis exerted beneficial effects in the dose of 60 mg/sheep/day, since it showed 83.3% of curative efficacy in the test CMT. This effect of curcumin can be direct or indirect, since the curcumin provided in the diet showed anti-inflammatory properties byreducing oxidation reactions and cell damage. Overall, the addition of curcumin in sheep diet improved the hematological and biochemical variables after only ten days of treatment. Future studies should be performed to clarify and determine whether the curcumin provided in diet can be an alternative to treat clinical or subclinical mastitis.Keywords: curcumin, mammary infection, immunity, biochemistry parameters, ovine.

Relationships among quarter milk leukocyte proportions and cow and quarter-level variables under different intramammary infection statuses.

The use of milk leukocyte differential (MLD) test has been proposed as a complement to somatic cell count (SCC) to assess the presence and the severity of intramammary infection. However, detailed information regarding the behavior of MLD under different physiological or pathological stages of the cow is nonexistent. The objective was to analyze the association between milk leukocyte proportions provided by a commercial automated MLD test and multiple cow and quarter-level variables. The study population consisted of 104 Holstein cows (32 primiparous and 72 multiparous) in one farm. Cows were categorized by days in milk as early (<50 DIM; n=29), middle (50–250 DIM; n=25), and late lactation (>250 DIM; n = 50). Milk from 416 quarters was collected and analyzed for lymphocytes (LYM), neutrophils (NEU), and macrophages (MAC) counts using an automated milk fluorescence microscopy system. Concurrently, a sterile composite milk sample was collected from each cow for pathogen identification through microbiological culture. Culture results were classified as no growth (NOG), gram-negative (GN), gram-positive (GP), or other (OTH). Milk leukocyte proportions varied depending on the level of total leukocyte counts (TLC; P < 0.001). Similarly, leukocyte ratios (NEU:LYM, NEU:MAC, and phagocyte:LYM) were different for multiple TLC categories (P < 0.05). There was no association between parity number and MLD; however, cows in early lactation had the greatest proportions of NEU and LYM. Leukocyte ratios varied depending on parity number and stage of lactation. Cows in the medium milk-yield category had the smallest proportions of NEU and LYM, and there was significant variation in leukocyte ratios, depending on the level of milk yield. In healthy quarters, MLD were not associated with quarter position; however, the NEU:MAC ratio was greater in rear quarters than in front quarters. In quarters with TLC >100,000, NEU% was greater in rear quarters than in front quarters (P = 0.03). For quarters with pathogen growth, TLC was greatest for GN followed by OTH and GP (P < 0.001). Milk LD depended on the isolated pathogen group, although the magnitudes of the differences were small. Although the changes in the proportions of leukocytes in milk were associated with categories of TLC, levels of milk yield, and mastitis-causing pathogen groups, the deviations were small in magnitude. Additional research is necessary to determine the potential applications for this methodology.

Prevalence of mastitis pathogens in South African pasture-based and total mixed ration-based dairies during 2008 and 2013.

Recent years have seen a change in the relative prevalence of environmental and contagious intramammary pathogens, as well as a change in the relative number of total mixed ration (TMR)-based and pasture (PAS)-based dairies in South Africa. The objectives of the study were to determine and compare the prevalence of mastitis pathogens in TMR and PAS dairies in South Africa during 2008 and 2013; furthermore, the within-herd prevalence of Streptococcus uberis in Str. uberis-positive herds was determined and compared. The prevalence of each pathogen, as well as the within-herd prevalence of Str. uberis, were compared between the two years and the two management systems using bacterial culture results from routinely collected composite cow milk samples submitted to the Onderstepoort Milk Laboratory, Faculty of Veterinary Science, University of Pretoria. Coagulase-negative staphylococci had the highest prevalence in both TMR and PAS dairies for both 2008 (29.60% [95.00% CI: 28.80% – 30.40%] and 26.90% [95.00% CI: 25.50% – 28.30%], respectively) and 2013 (20.20% [95.00% CI: 19.30% – 21.10%] and 22.70% [95.00% CI: 22.20% – 23.10%], respectively), which decreased significantly from 2008 to 2013 in both TMR and PAS dairies (p < 0.001). Streptococcus uberis showed an increase in prevalence in both TMR (p = 0.002) and PAS dairies (p = 0.001) from 2008 (2.36% [95.00% CI: 2.10% – 2.65%] and 2.63% [95.00% CI: 2.16% – 3.16%], respectively) to 2013 (3.10% [95.00% CI: 2.72% – 3.51%] and 3.64% [95.00% CI: 3.45% – 3.83%], respectively). Staphylococcus aureus showed a significant decrease in both TMR (p = 0.011) and PAS (p < 0.001) dairies from 2008 (4.71% [95.00% CI: 4.34% – 5.10%] and 5.62% [95.00% CI: 4.94% – 6.36%], respectively) to 2013 (3.95% [95.00% CI: 3.52% – 4.40%] and 1.71% [95.00% CI: 1.58% – 1.84%], respectively). The median within-herd prevalence of Str. uberis for the combined dairy systems showed a significant increase from 2008 (1.72% [IQR: 0.88% – 5.00%]) to 2013 (3.10% [IQR: 1.72% – 4.70%]) (p < 0.001). Statistically significant differences were found in the prevalence of most of the major contagious and environmental mastitis pathogens between 2008 and 2013 and between TMR and PAS dairies. The within-herd prevalence of Str. uberis increased from 2008 to 2013, with the highest within-herd prevalence in PAS dairies in 2013.

Use of MALDI-TOF to characterize staphylococcal intramammary infections in dairy goats

The most common pathogens causing intramammary infections (IMI) in dairy goats are staphylococci. Gene sequencing has been the reference method for identification of staphylococcal species, but MALDI-TOF mass spectrometry could represent a rapid and cost-effective alternative method. The objectives were to evaluate the typeability and accuracy of partial gene sequencing and MALDI-TOF for identifying staphylococci isolated from caprine milk samples, and to evaluate the relationship between staphylococcal species IMI, milk somatic cell score (SCS), and milk yield (MY). A composite (goat-level) milk sample was collected from all 940 lactating goats in a single herd. Dairy Herd Information Association test-day data for parity, days in milk, SCS, and MY were retrieved from Dairy Herd Information Association records. Milk samples were cultured on Columbia blood agar, and isolates from samples that yielded a single colony type of a presumptively identified Staphylococcus spp. were identified by PCR amplification and partial sequencing of rpoB, tuf, or 16S-rRNA, and MALDI-TOF. Mixed linear models were used to evaluate the relationship between staphylococcal IMI, SCS, and MY. The goat-level prevalence of staphylococcal IMI based on isolation of a single colony type was 24.4% (213/874). Seventeen goats had a contaminated sample. Among the remaining goats (n = 857), the most common species causing single colony-type IMI were Staphylococcus simulans (7.9%), Staphylococcus xylosus (3.5%), Staphylococcus caprae (3.6%), Staphylococcus chromogenes (2.9%), and Staphylococcus epidermidis (2.2%). The typeability of staphylococcal isolates with partial housekeeping gene sequence analysis (rpoB, complemented by tuf and 16S as needed) was 97.7%. The typeability and accuracy of MALDI-TOF were 84 and 100%, respectively. Overall, only Staphylococcus chromogenes IMI was associated with a higher SCS than goats with no growth. After adjusting for parity and stage of lactation, staphylococcal IMI status was not significantly associated with MY. For the staphylococci isolated from goats in this herd, MALDI-TOF proved an accurate method of speciation with a relatively high typeability. An association between staphylococcal IMI, SCS, and MY was not defined using goat-level data with the exception of S. chromogenes IMI, which was associated with a higher SCS than goats with no growth.

Influence of sampling technique and bedding type on the milk microbiota: Results of a pilot study

The objective of this pilot study was to evaluate the influence of sampling technique and exposure to different bedding types on the milk microbiome of healthy primiparous cows. Primiparous Holstein cows (n = 20) with no history of clinical mastitis or monthly somatic cell counts >150,000 cells/mL were selected for this study. From each enrolled cow, a composite milk sample was aseptically collected from all 4 mammary quarters (individual quarter somatic cell counts <100,000 cells/mL), 1 individual quarter milk sample was collected using conventional aseptic technique, and 2 individual quarter milk samples were collected directly from the gland cistern using a needle and vacuum tube. All milk samples were cultured using standard milk microbiological techniques and DNA was extracted. Extracted DNA was subjected to PCR and next-generation sequencing for microbiota determination. All samples yielded relatively little total DNA. Amplification of PCR was successful in 45, 40, and 83% of composite, conventional, and cisternal samples, respectively. Bacteria were successfully cultured from 35% of composite milk samples but from none of the quarter milk samples collected using conventional or cisternal sampling techniques. Bacterial DNA sequences were assigned to operational taxonomic units (OTU) based on 97% sequence similarity, and bacterial richness and diversity were determined. Most samples were dominated by low-prevalence OTU and of the 4,051 identified OTU, only 14 were prevalent at more than 1% each. These included bacteria typically recovered from environmental sources. Chao richness was greatest in composite samples and was 636, 347, and 356 for composite, conventional quarter, and cisternal milk samples, respectively. Shannon diversity was similar among sample types and ranged from 3.88 (quarter) to 4.17 (composite). Richness and diversity did not differ by bedding type among cisternal samples, but the power of this pilot study was limited due to small sample size. Despite the small sample size, for milk samples collected from the gland cistern, overall bacterial community composition differed among bedding types. These results demonstrate that sampling technique and bedding type may be associated with the microbiota detected in bovine milk, and we suggest that these variables should be considered in designing and reporting studies about the milk microbiota.

Latent class analysis of real time qPCR and bacteriological culturing for the diagnosis of Streptococcus agalactiae in cow composite milk samples

Streptococcus agalactiae (S. agalactiae) has re-emerged as a mastitis pathogen among Norwegian dairy cows. The Norwegian cattle health services recommend that infected herds implement measures to eradicate S. agalactiae, this includes a screening of milk samples from all lactating cows. The performance of the qPCR-test currently in use for this purpose has not been evaluated under field conditions. The objective of this study was to estimate the sensitivity and specificity of the real-time qPCR assay in use in Norway (Mastitis 4 qPCR, DNA Diagnostics A/S, Risskov, Denmark) and compare it to conventional bacteriological culturing for detection of S. agalactiae in milk samples. Because none of these tests are considered a perfect reference test, the evaluation was performed using latent class models in a Bayesian analysis. Aseptically collected cow-composite milk samples from 578 cows belonging to 6 herds were cultured and tested by qPCR. While 37 (6.4%) samples were positive for S. agalactiae by bacteriological culture, 66 (11.4%) samples were positive by qPCR. The within-herd prevalence in the six herds, as estimated by the latent class models ranged from 7.7 to 50.8%. At the recommended cut-off (cycle threshold 37), the sensitivity of the qPCR was significantly higher at 95.3 (95% posterior probability interval [PPI] [84.2; 99.6]) than that of bacteriological culture at 58.2 (95% PPI [43.8; 74.4]). However, bacterial culture had a higher specificity of 99.7 (95% PPI [98.5; 100.0]) compared to the qPCR at 98.5 (95% PPI [94.6; 99.9]). The median estimated negative predictive values of qPCR was consistently higher than those of the BC at all estimated prevalences, and the superiority of the qPCR increased with increasing within-herd prevalence. The median positive predictive values of BC was in general higher than the estimates for the qPCR, however, at the highest prevalence the predictive ability of both tests were similar.

Prediction of sheep milk chemical composition using milk yield, pH, electrical conductivity and refractive index.

This Research Communication addresses the hypothesis that fat, protein, lactose and total solids content can be predicted using daily milk yield (DMY), pH, electrical conductivity (MEC) and refractive index (RI) of milk as predictors. It also addresses the possibility of these measurements being used for on-farm benchmarking activities towards selecting the highest yielding animals and flocks regarding milk quality traits (MQT). A total of 308 purebred Frizarta ewes were used for the study. From each individual ewe, a composite milk sample was collected. pH, MEC and RI of milk were measured and the samples were assayed for fat, protein, lactose and total solids content, using an automatic infrared milk analyser. The predictive value of DMY, pH, MEC and RI of milk on its MQT was assessed using multiple linear regression analysis. Significant regression equations were produced for all of the studied traits. RI and MEC were significant and reliable predictors for all studied MQT, whereas DMY was a significant predictor for most MQT with the exception of protein content. pH was a marginally significant predictor for some of the MQTs at the initial development of the equations but proved unreliable after bootstraping. Using these equations a number of ewes varying from 75 (for fat) to 97 (for protein) out of the 100 highest MQT yielders were correctly predicted, whereas, none of the ewes out of the 100 lowest MQT yielders was mispredicted as a high yielder for protein-, lactose- and total solids- content. Three out of 100 lowest fat-yielders were mispredicted as high fat-yielders. Similar equations can be used for benchmarking activities towards selecting the highest protein-, fat-, lactose- and total solids- yielding animals and flocks in cases where laboratories for MQT analyses are not readily available or the cost of chemical analyses is high. The method can be regarded as a handy tool for the dairy industry to readily assess milk quality at the farm level.

Review on California Mastitis Test (CMT)

The California Mastitis Test (CMT) is a quick, simple test that accurately predicts the somatic cell count of milk from individual quarters or on composite milk samples [1]. The California Mastitis Test is a simple, inexpensive, rapid screening test for mastitis. The test is based upon the amount of cellular nuclear protein present in the milk sample. Since inflammatory cells associated with mastitis are the predominant cell type present in milk the CMT reflects the SCC level quite accurately and is a rel iable indicator of the severity of infection. The test is appropriate for cow - side evaluation of udder health and the procedure can be taught quickly to producers and the milking crew. With proper training test results are very repeatable among those condu cting the test. It is necessary that producers also be made aware of the limitations of the CMT and the proper application of management decisions based on CMT results.

Prevalence and bacterial etiology of subclinical mastitis in goats reared in organized farms.

Aim:Assessment of the status of subclinical mastitis (SCM) in Jamunapari and Barbari goats in Indian organized farms, the involvement of bacterial pathogens and their sensitivity to antibiotics.Materials and Methods:A total of 181 composite milk samples were aseptically collected from the apparently healthy Barbari (n=95) and Jamunapari (n=86) goats. The California mastitis test (CMT) and somatic cell count (SCC) were used to diagnose SCM. The milk samples with CMT scores of 0 and +1 were considered as negative, while the samples with the score of +2 or +3 were taken as positive, and further, the positive samples were used for the bacteriological examination. An antibiotic sensitivity test was performed by disk diffusion method using seven commercially available antibiotic discs.Results:All the samples having CMT score of +2 or +3 demonstrated SCC more than 1 million. Overall, the prevalence of SCM in the goats was assessed as 19.89% (36/181). The prevalence of SCM in Barbari and Jamunapari goats was found as 24.21% (23/95) and 15.12% (13/86), respectively. Out of 11 isolates of Staphylococci, 9 isolates were identified as coagulase-negative Staphylococci (CNS), whereas 2 isolates were found as Staphylococcus aureus. The identified bacterial isolates (n=30) did not show antibiotic resistance.Conclusion:The current investigation showed the considerable prevalence of SCM among Jamunapari and Barbari goats which may have a negative impact on quantity and quality of the milk. CNS was found as the most prevalent cause of SCM in the goats. Negligible antibiotic resistance was found among the identified udder pathogens.

Prepartum application of internal teat sealant or intramammary amoxicillin on dairy heifers: Effect on udder health, survival, and performance

Intramammary infusion of antimicrobials (IA) and application of an internal teat sealant (ITS) at the end of the lactation is a common strategy to prevent and treat intramammary infections during the dry period. In heifers, precalving IA has been reported to improve udder health and milk quality; however, the effect of ITS in heifers housed in freestall barns is still unknown. The objective of this study was to investigate the effect of prepartum application of ITS or IA on udder health of dairy heifers. A total of 886 heifers were randomly allocated into 1 of 4 treatment groups when they were 245 ± 3 d pregnant. Control heifers (CON) did not receive any treatment. Heifers in the ITS group received the application of ITS containing 2.6 g of bismuth subnitrate. Animals in the IA group received intramammary administration of 62.5 mg of amoxicillin. Heifers in IA+ITS group received intramammary administration of amoxicillin followed by ITS application. The follow-up period was 270 d in milk (DIM). Clinical mastitis was diagnosed and treated by trained farm personnel throughout the study period. Composite milk somatic cell counts (SCC) were determined monthly for the first 9 mo of lactation by Dairy Herd Improvement Association. Subclinical mastitis was defined as a cow having a SCC >200,000 cells/mL for at least 1 test day and not diagnosed with clinical mastitis during the study period. At 7 ± 3 DIM, composite milk samples were collected for bacteriological analysis for a subset of the study animals. The presence of pathogens in milk at 7 ± 3 DIM was more frequently detected in the CON heifers compared with animals enrolled in the treatment groups. Heifers in IA+ITS group had a decreased incidence of clinical mastitis compared with CON heifers (12.9 vs. 21.4%). Additionally, the incidence of subclinical mastitis was decreased for IA+ITS heifers compared with CON counterparts (20.1 vs. 43.8%). During the first 9 mo of lactation, IA and IA+ITS heifers had lower SCC linear scores compared with ITS and CON counterparts. Although IA+ITS treatment successfully improved udder health, it did not result in better milk yield, fertility, and survivability. In conclusion, ITS alone was not effective in improving udder health; however, the combination of ITS and IA decreased the incidence of clinical and subclinical mastitis during the first 9 mo of lactation. Additionally, IA+ITS had a long-term effect on the SCC linear score throughout the study period.

Association of milk quality parameters with teat and udder traits in Tharparkar cows

The present experiment was carried out to study the association of milk quality traits (fat, protein, lactose, solid not fat (SNF), total solids (TS) and somatic cell count (SCC)) with teat traits ((teat length (TL), teat diameter (TD), distance between teats (DbT) and teat tip height from ground (THFG)) and udder traits ((udder circumference= UC, udder depth (UD), udder width (UW) and udder height from ground (UHFG)), in Tharparkar cows. The experiment was carried out on 32 tharparkar cows of different parities and lactation stages. Morning and evening measurement of teat and udder traits was carried out before milking and approximately 30 ml of representative milk samples were taken from each of the functional quarters and one composite milk sample of each experimental animal fortnightly from the date of calving of the animal in clean sampling bottles. Fat% and SNF% showed a negative, significant (P&gt;0.05) to highly significant (P&gt;0.01) correlation with TD, UL and FUD. Protein% expressed a positive, significant (P&gt;0.05) to highly significant (P&gt;0.01) correlation with UC, FUD and DbT. Further, Fat%, SNF% and TS% expressed a positive, significant (P&gt;0.05) to highly significant (P&gt;0.01) correlation with THFG. SCC (×105/ml) showed a positive, significant (P&gt;0.05) to highly significant (P&gt;0.01) correlation with TL, TD, DbT, UC and UL, whereas it had a negative correlation with THFG, rear-udder depth (RUD) and UW. It was further observed that Fat% expressed a positive highly significant (P&gt;0.01) phenotypic correlation with SNF% and TS%. SNF%, Lactose%, Protein% and TS% expressed a negative phenotypic correlation with SCC.

Relationship between intramammary infection prevalence and somatic cell score in commercial dairy herds

We examined consistency of the relationship between intramammary infection (IMI) and somatic cell score (SCS) across several classes of cow, herd, and sampling time variables. Microbial cultures of composite milk samples were performed by New York Quality Milk Production Services from 1992 to 2004. SCS was from the most recent Dairy Herd Improvement test before IMI sampling. Records were analyzed from 79,308 cows in 1,124 commercial dairy herds representing a broad range of production systems. Three binary dependent variables were presence or absence of contagious IMI, environmental IMI, and all IMI. Independent variables in the initial models were SCS, SCS2, lactation number, days in milk, sample day milk yield, use of coliform mastitis vaccine, participant type (required by regulation or voluntary), production system (type of housing, milking system, and herd size), season of sampling, year of sampling, and herd; also the initial models included interactions of SCS and SCS2 with other independent variables, except herd and milk yield. Interaction terms characterize differences in the IMI-SCS relationship across classes of the independent variables. Models were derived using the Glimmix macro in SAS (SAS Institute Inc., Cary, NC) with a logistic link function and employing backward elimination. The final model for each dependent variable included all significant independent variables and interactions. Simplified models omitted SCS2 and all interactions with SCS. Interactions of SCS with days in milk, use of coliform mastitis vaccine, participant type, season, and year were not significant in any of the models. Interaction of SCS with production system was significant for the all IMI model, whereas interaction of SCS with lactation number was significant for the environmental and all IMI models. Each 1-point increase in SCS (or doubling of somatic cell count) was associated with a 2.3, 5.5, and 9.1% increase in prevalence of contagious, environmental, and all IMI, respectively. Empirical receiver operator characteristic curves and areas under the curve were derived for final and simplified models. The areas under the curve for simplified and final models within each type of IMI differed by 0.009 or less. We concluded that the relationship of IMI with SCS was generally stable over time and consistent across seasons, production systems, and cow factors.

The association between subclinical mastitis around calving and reproductive performance in grazing dairy cows

The objective of this study was to evaluate the effect of subclinical mastitis (SCM) on calving-to-first-service interval (CFS), calving-to-conception interval (CC), and on the number of services per conception (S/C) in grazing Holstein and Normande cows. Primiparous (n=43) and multiparous (n=165) cows were selected from five dairy herds. Two composite milk samples were aseptically collected from each cow at drying-off, and then every week during the first postpartum month. One sample was used for somatic cell count (SCC), and the other one for bacteriological analysis. Cows were followed up to 300 d after calving. Non-parametric and parametric survival models, and negative binomial regression were used to assess the association between SCM, evaluated by SCC and milk culture, and reproductive indices. Staphylococcus aureus, CNS, and Streptococcus uberis were the most frequent isolated pathogens. Subclinical mastitis in the first month of lactation was not associated with CFS; however, the CC interval was longer in cows with SCM compared to healthy cows, the former also had a higher number of S/C.

Prevalence of Subclinical Mastitis and Distribution of Pathogens in Dairy Farms of Rubavu and Nyabihu Districts, Rwanda.

A cross-sectional study was conducted from May 2016 to January 2017 in Rubavu and Nyabihu districts, Western Rwanda, aiming at estimating the prevalence of subclinical mastitis (SCM) and identifying its causative bacteria. Management practices and milking procedures were recorded through a questionnaire. 123 crossbreed milking cows from 13 dairy farms were randomly selected and screened for SCM using California Mastitis Test (CMT). Composite CMT positive milk samples were processed for bacterial isolation and identification. The overall SCM prevalence at cow level was 50.4%. 68 bacterial isolates were identified by morphological and biochemical characteristics. They included, Coagulase Negative Staphylococci (51.5%), Staphylococcus aureus (20.6%), Streptococcus species (10.3%), Bacillus species (10.3%), Streptococcus agalactiae (5.8%), and Escherichia coli (1.5%). About 67.1% of the farmers checked for mastitis; of these, 58.9% relied on clinical signs and only 6.8% screened with CMT. Only 5.5% and 2.7% of the farmers tried to control mastitis using dry cow therapy and teat dips, respectively. Thus, to reduce the prevalence of SCM, farmers in the study area need to be trained on good milking practices, including regular use of teat dips, application of dry cow therapy, and SCM screening. This will improve their sales and their financial status.