Indicator Of Subclinical Mastitis Research Articles

Somatic cell count as an indicator of subclinical mastitis and increased inflammatory response in asymptomatic lactating women.

This pilot study suggests that SCC at a level of (greater than or equal to) 250,000 cells mL-1, as used in the dairy industry, is a suitable index to identify asymptomatic subclinical mastitis in lactating women since it reflects a significant increase in the inflammatory response compared to milk samples from healthy women. Using this index should aid studies into the short- and long-term consequences of subclinical mastitis for mother and infant.

162 Evaluating the Potential of a Thermal Imaging System to Identify Subclinical Mastitis in Dairy Cattle

Abstract The objective was to evaluate the potential for the use of a thermal imaging system for the identification of subclinical mastitis in dairy cows. Multiparous Holstein dairy cows (n = 131) within 150 days in milk (DIM; range 65 to 155) were enrolled in the study at a commercial dairy farm in central Texas. Milk samples (10 mL) were collected from each quarter separately and evaluated for somatic cell counts (SCC; n*1000), fat (%), protein (%), lactose (%), SNF (%), BHBA (mM/L), acetone (mM/L), and milk urea nitrogen (MUN; mg/dL). A minimum of four thermal images per cow representing all four quarters were captured using a handheld infrared camera (Fluke TiS 75+) from approximately one-meter distance from the cow. All images were taken inside the milking parlor and after the morning milking during February 2023 to minimize the impact of daily ambient temperatures. Skin temperatures from each quarter (maximum, minimum, average) were extracted using Fluke Connect software. A total of 524 quarter-level milk samples and corresponding udder images were obtained, and 10 images were discarded from analyses due to the lack of appropriate resolution and focus. To prevent collinearity, a correlation matrix was developed using PROC CORR in SAS and variables with significant correlations were removed from the model. Subclinical mastitis was defined at the quarter level when SCC > 250,000. Statistical analyses were performed using mixed linear regression models (PROC MIXED) with cow and DIM as random effects in the model, and logistic regression model (PROC LOGISTIC) was fitted to evaluate the probability of the disease based on temperature and milk variables. Maximum and average udder temperatures did not demonstrate a significant relationship with subclinical mastitis status whereas minimum udder temperature was greater in sick quarters (29.6 ± 0.52 vs 27.5 ± 0.45; P = 0.003). Increased minimum udder temperature was also associated with BHBA (P < 0.0001), acetone (P < 0.0004), and MUN (P < 0.0007) concentrations in milk. Cows with subclinical mastitis had decreased BHBA (0.14 ± 0.037 vs 0.28 ± 0.031; P = 0.0036) and acetone (0.017 ± 0.003 vs 0.044 ± 0.003; P < 0.001). Odds of subclinical mastitis were associated with MUN values in milk (OR 0.94; CI 0.89 - 0.98) but not with any udder surface temperature variables. We conclude that an increase in the minimum temperature of the udder could be used as an indicator of subclinical mastitis but studies with larger datasets and using advanced image analysis techniques are warranted to validate these results.

The importance of somatic cell count in dairy technology

Mastitis is an important infection that affects herd health, udder health and milk yield of individual animals. Somatic cells (SCs) naturally present in milk are used as an indicator of subclinical mastitis, but also to assess milk quality and safety. Somatic cell count is the main monitoring variable used in milk quality and safety assessment. This review especially focuses on the effect of SCs on milk as food and also their effect on the dairy technology. The most significant effect of SCC on dairy technology is provided by the enzymes released from these cells. Alternatively, a limited number of conclusions were reported that SCs may assist dairy technology. SCs pass to milk and continue inhibitory effect on bacteria.

Chemometric Approaches to Analyse the Composition of a Ewe's Colostrum.

Colostrum is a major source of immunity in lambs and, in general, in all newborn ruminants. It allows the transfer of antibodies from the ewe to the lamb, and it becomes the exclusive source of nutrients for a newborn. Among the most significant Pearson correlations, the positive correlation between the Brix refractive index (Brix) and protein (0.90) should be noted. Both parameters (protein percentage and Brix) were then positively correlated with the percentage of fat (0.38 and 0.41), urea (0.81 and 0.67), calcium (0.39 and 0.29), and magnesium (0.58 and 0.59), as well as the yellowness (0.78 and 0.75). Somatic cell count (SCC) and pH, parameters which are indicators of subclinical mastitis, were positively correlated (0.49), and SCC was positively correlated with sodium (0.37) and negatively correlated with potassium (-0.28). Among the macroelements in colostrum-calcium, potassium, magnesium, and sodium-the correlations were largely positive. With respect to the colour parameters, yellowness was negatively correlated with lightness (-0.41) and redness (-0.45). The factor analysis split the total of variance into three latent factors. The first factor was named "Colostrum quality of grazing sheep" because it was positively correlated primarily with SCC, pH, Poly Unsatured Fatty Acids (PUFA), and the sum of the omega-3 Fatty Acids (FAs). The second factor, named "Good quality colostrum", was positively correlated primarily with the refractive index, protein and fat percentages, urea content, phosphorus, Mono Unsatured Fatty Acids (MUFA), and yellowness. The third factor was positively correlated primarily with calcium, potassium, magnesium, and sodium, and it was therefore termed "Mineral component of colostrum". Stepwise discriminant analysis showed that the protein percentage, calcium, and magnesium were able to explain more than 85% of the Brix refractive index, which remains a good parameter for simply evaluating the nutritional quality of sheep colostrum at the level of a farm.

Epidemiological investigation of mastitis in lactating buffalo cows in Nineveh governorate, Iraq

The study's objective was to investigate the incidence and predisposing factors of mastitis in lactating buffalo cows in the Nineveh governorate. A total of 160 recently calved buffalo cows were followed up in a prospective study during the first three months of lactation. Cases of clinical mastitis (CM) were investigated, and milk was sampled to identify the increase in somatic cell count (SCC) as an indicator of subclinical mastitis (SCM). Several epidemiological factors that can predispose to mastitis were also investigated. The incidence of CM was 9.4%, mostly recorded during the first ten days (4 days on average). The average CM duration per case was 35 ± 4.9 days. Different changes and clinical findings were revealed in milk and affected udder. The incidence of SCM (i.e., SCC > 200,000 per mL) was 83.75%, mostly recorded during the first month of lactation. Furthermore, the odds of SCM were high in winter. The study concluded that the incidence of CM in buffalo cows is very low. The chronic form characterizes CM, SCM is more common than CM, and winter is a predisposing factor for the infection. Examination of several control measures of mastitis is important to improve the production and hygiene of buffalo milk in Nineveh governorate.

Relationship between total and differential quarter somatic cell counts at dry-off and early lactation.

Mastitis is a most common disease of dairy cows and causes tremendous economic loss to the dairy industry worldwide. Somatic cell counts (SCC) reflect the inflammatory response to infections and is a metric used as key indicator in mastitis screening programs, typically within the framework of national milk recording schemes. Besides the determination of total SCC, the differentiation of cell types has been described to be beneficial for a more definite description of the actual udder health status of dairy cows. Differential somatic cell count (DSCC) represents the combined proportion of polymorphonuclear leukocytes (PMN) and lymphocytes expressed as a percentage of the total. The aim of this study was to investigate the relationship between SCC and differential somatic cell count (DSCC) in individual quarter milk samples collected at different time points: at dry-off, after calving and at the lactation peak. We used individual quarter data from farms representing the specialized production system of Parmigiano Reggiano cheese in Northern Italy. Average DSCC values ranged between 44.9% and 56.3%, with higher values (60.4%-72.1%) in milk samples with ≥ 1 million SCC/ml (where the proportion of samples with DSCC > 70% can be as high as 0.73). Moderate overall correlations between DSCC and log(SCC) were estimated (Pearson = 0.42, Spearman = 0.38), with a clear increasing trend with parity and around the lactation peak (e.g. Pearson = 0.59 at 60 DIM in parity 4). Taking SCC values as indicators of subclinical mastitis, DSCC would diagnose mastitis with 0.75 accuracy. Data editing criteria do have an impact on results, with stricter filtering leading to lower correlations between log(SCC) and DSCC. In conclusion DSCC and SCC provide different descriptions of the udder health status of dairy cows which, at least to some extent, are independent. DSCC alone doesn’t provide more accurate information than SCC at quarter level but, used in combination with SCC, can be of potential interest within the framework of milk recording programs, especially in the context of selective dry-cow therapy (SDCT). However, this needs further investigation and updated threshold values need to be selected and validated.

Infant Anthropometry and Growth Velocity Before 6 Months are Associated with Breastfeeding Practices and the Presence of Subclinical Mastitis and Maternal Intestinal Protozoa in Indigenous Communities in Guatemala.

ABSTRACTBackgroundThe possibility that maternal health status and breastfeeding practices contribute to growth faltering before 6 mo is underexplored.ObjectivesThis longitudinal study investigated whether indicators of subclinical mastitis (SCM) and breast inflammation, maternal fecal-oral contamination, and/or breastfeeding practices were associated with infant anthropometry or growth velocity before 6 mo.MethodsIndigenous Mam-Mayan mother-infant dyads (n = 140) were recruited. Breast milk was collected at early (<6 wk) and established (4–6 mo) lactation when maternal and infant anthropometry were measured. Milk Na:K ratio as an indicator of SCM and concentrations of 4 proinflammatory cytokines were measured. Maternal stool samples were examined for the presence of intestinal parasites including nonpathogenic protozoa (Endolimax nana, Iodamoeba bütschlii, Entamoeba coli, Blastocystis hominis). Questionnaires characterized breastfeeding and hygiene practices. Multiple linear regression identified factors associated with infant growth attainment [weight-for-age z-score (WAZ), length-for-age z-score (LAZ), and head circumference-for-age z-score (HCAZ)] and growth velocity (expressed as change per day from early to established lactation). Multiple logistic regression identified factors associated with increased odds of underweight, stunting, and low head circumference at both lactation stages.ResultsA higher Na:K ratio, individual nonpathogenic protozoa, and specific breastfeeding and hygiene practices were associated with impaired growth before 6 wk and at 4–6 mo in exclusively breastfed infants. Growth velocity for weight was inversely associated with Entamoeba coli but cranial growth was associated positively with Iodamoeba bütschlii whereas feeding colostrum in early lactation was protective and decreased the odds of an HCAZ < −2 SD. Finally, the presence of SCM in early lactation increased the likelihood of both WAZ < −2 SD and LAZ < −2 SD by 6 wk.ConclusionsPrevention of SCM can improve early infant weight, but measures that promote the feeding of colostrum and reduce exposure to fecal-oral contamination might be required to minimize infant growth faltering in breastfed infants.

Application of udder surface temperature by infrared thermography for diagnosis of subclinical mastitis in Holstein cows located in tropical highlands

Several reports have indicated that udder surface temperature (UST) can be a useful indicator of subclinical mastitis (SCM). The objective was to evaluate UST by infrared thermography (IRT) as a diagnostic tool for SCM and intramammary infection (IMI), and to assess the influence of environmental conditions in the potential diagnosis of this disease in dairy cows located at high-altitude tropical regions. A total of 105 cows (397 quarters) from 3 dairy farms with mechanical and manual milking methods were enrolled in the study. Subclinical mastitis was diagnosed when quarter samples had a somatic cell count (SCC) ≥200 × 103 cells/mL, microbial growth (MG) was defined when a major pathogen (≥1 cfu/plate) or Corynebacterium spp. (≥10 cfu/plate) was isolated, and IMI was defined as the presence of MG and SCC ≥100 × 103 cells/mL. Infrared images were taken with a thermal camera placed 1 m away from the udder, and shots of the rear and left and right lateral view were made during the morning milking, before any manipulation of the udder and employing dark cardboard on the contralateral side to avoid artifacts in the background. A multilevel mixed effects linear regression model clustered within cows and herd was performed to evaluate the associations with UST. Clinical performance was evaluated using the Youden index to establish the optimum UST thresholds, which were set at 32.6°C for any case definition when milking was by hand, at 33.7°C for MG, and at 34°C for SCM and IMI in machine-milked quarters. Sensitivity (Se), specificity (Sp), area under curve (AUC), and positive likelihood ratio (+LR) were also assessed. Test agreement was assessed by kappa coefficient (κ). The UST of healthy quarters ranged between (95% CI) 32.4 and 32.6°C, lower than SCM quarters (n = 88) at 32.9°C (95% CI: 32.7-33.1 °C), MG quarters (n = 56) at 33.5°C (95% CI: 33.3-33.7°C), and IMI quarters (n = 50) at 33.5°C (95% CI: 33.2-33.7 °C). The UST was also related to the milking method: higher temperatures were observed for hand milking (n = 90) compared with machine milking (n = 185). No relation between environmental conditions such as wind speed, atmospheric temperature, relative humidity, and temperature-humidity index and UST were observed during this study. For hand milking, the optimal UST threshold was 32.6°C; for SCM, Se = 0.53, Sp = 0.89, AUC = 0.71, κ = 0.4; for MG, Se = 0.83, Sp = 0.93, AUC = 0.88, κ = 0.77; and for IMI, Se = 0.82, Sp = 0.92, AUC = 0.87, κ = 0.74. The machine milking threshold for SCM resulted in Se = 0.42, Sp = 0.97, AUC = 0.70, κ = 0.47; for MG, Se = 0.82, Sp = 0.89, AUC = 0.85, κ = 0.60; and for IMI, Se = 0.82, Sp = 0.98, AUC = 0.90, κ = 0.79. These findings suggest that UST determined by IRT is higher in machine-milked cows and in quarters with MG and IMI than in healthy quarters; therefore, UST by IRT is a reliable, clinically useful method for MG and IMI diagnosis.

Associations of freestall design and cleanliness with cow lying behavior, hygiene, lameness, and risk of high somatic cell count

The objective of this study was to investigate associations of freestall design and cleanliness with cow lying behavior, hygiene, lameness, and risk of new high somatic cell count (SCC). Cows from 18 commercial freestall dairy herds (22 ± 15 cows/farm; mean ± SD) in Ontario, Canada, were enrolled in a longitudinal study. Four hundred focal cows that were <120 d in milk, had no mastitis treatment in the last 3 mo, and had an SCC <100,000 cells/mL at their most recent milk test were selected for the study. Data on SCC were collected through Dairy Herd Improvement Association milk testing (at ~5-wk intervals). Each farm was visited 5 ± 3 d (mean ± SD) after each milk test until 3 tests were completed (~105 d), for a total of 3 observation periods per cow. Elevated SCC was used as an indicator of subclinical mastitis. An incident of new high SCC was defined as a cow having SCC >200,000 cells/mL at the end of an observation period, when SCC was <100,000 cells/mL at the beginning of that period. Lying behavior was recorded for 6 d after each milk sampling, using electronic data loggers. Cows were scored during each period for lameness (5-point scale, with scores ≥3 = lame), body condition score (BCS; 5-point scale; 1 = thin to 5 = fat), and hygiene (4-point scale). Stall cleanliness was assessed during each period with a 1.20 × 1.65-m metal grid, containing 88 squares. The grid was centered between stall partitions of every tenth stall on each farm, and the squares containing visible urine or fecal matter (or both) were counted. Cow lying time averaged 10.9 ± 1.9 h/d. On average, cows with low BCS (≤2.5) spent 37 ± 16.6 min/d less time lying down than high-BCS cows (≥4.0). On average, cows tended to spend 36 ± 18.3 min/d more time lying down in deep-bedded versus mattress-based stalls. Mean proportion of soiled squares per stall was 20.1 ± 0.50%. Across farms, cow lying time decreased as the proportion of soiled squares per stall increased. A difference in daily lying time of ~80 more min/d was modeled for cows housed in barns with the cleanest stalls compared with those with the dirtiest stalls. Higher neck rail height [for every 1 SD (10 cm) increase] increased the odds (odds ratio = 1.5) of cows having a dirty upper leg-flank and udder. The odds of a cow having a dirty upper leg-flank, udder, and lower legs were 1.5, 2.0, and 1.9 times greater, respectively, for cows housed with dirtier stalls. Also, cows housed on farms with dirtier stalls had 1.3 times greater odds of being lame at the time of observation. Over the study period, 50 new high-SCC cases were detected, resulting in an incidence rate of 0.45 cases of new high SCC per cow-year at risk. No measured factors were detected to be associated with risk of a new high SCC. Overall, our results confirm that cows lie down longer in cleaner and more comfortable environments. Further, these results highlight the need for improved stall cleanliness to optimize lying time and potentially reduce lameness.

Microbiota characterization of sheep milk and its association with somatic cell count using 16s rRNA gene sequencing.

This work aimed to use 16S ribosomal RNA sequencing with the Illumina MiSeq platform to describe the milk microbiota from 50 healthy Assaf ewes. The global observed microbial community for clinically healthy milk samples analysed was complex and showed a vast diversity. The core microbiota of the sheep milk includes five genera: Staphylococcus, Lactobacillus, Corynebacterium, Streptococcus and Escherichia/Shigella. Although there are some differences, some of these genera are common with the microbiota core pattern of milk from other species, especially with dairy cows. The microbial composition of the studied samples, based on the definition of amplicon sequence variants, was analysed through a correlation network. A preliminary analysis by grouping the milk samples based on their somatic cell count (SCC), which is considered an indicator of subclinical mastitis (SM), showed certain differences for the core of the samples identified as SM. The differences in the microbiota diversity pattern among samples might also suggest that subclinical mastitis would be associated with the significant increase in some genera that are inhabitants of the mammary gland and a remarkable concomitant reduction in the microbial diversity. Additionally, we have also presented here a preliminary analysis to assess the impact of the sheep milk microbiome on SCC, as an indicator of subclinical mastitis. The results here reported provide a first characterization of the sheep milk microbiota and settle the basis for future studies in this field.

Genetic analysis of subclinical mastitis in early lactation of heifers using both linear and threshold models

Subclinical mastitis (SCM) causes economic losses for dairy producers by reducing milk production and leading to higher incidence of clinical mastitis and premature culling. The prevalence of SCM in first-lactation heifers is highest during early lactation. The objective of this study was to estimate genetic parameters for SCM in early lactation in first-parity Holsteins. Somatic cell count test-day records were collected monthly in 91 Canadian herds participating in the National Cohort of Dairy Farms of the Canadian Bovine Mastitis Research Network. Only the first test-day record available between 5 and 30 d in milk was considered for analysis. The final data set contained 8,518 records from first lactation Holstein heifers. Six alternative traits were defined as indicators of SCM, using various cutoff values of SCC, ranging from 150,000 to 400,000 cells/mL. Both linear and threshold animal models were used. Overall prevalence of SCM using the 6 traits ranged from 13 to 24%. Heritability estimates (standard error) from linear and threshold models ranged from 0.037 to 0.057 (0.015 to 0.018) and from 0.040 to 0.051 (0.017 to 0.020), respectively. We found strong genetic correlations (standard error) among alternative SCC traits, ranging from 0.90 to 0.99 (0.013 to 0.069), indicating that these 6 traits were genetically similar. Despite low heritability, based on estimated breeding values (EBV) predicted from both models, we noted exploitable genetic variation among sires. Higher EBV of SCM resistance corresponded to sires with a higher percentage of daughters without SCM. Based on a linear model (all 6 traits), percentage of daughters with SCM ranged from 5 to 13% and from 19 to 33% for the top 10% and worst 10% of 69 sires with minimum 20 daughters in at least 5 herds, respectively. Spearman's rank correlations among EBV of sires predicted from linear (from 0.75 to 0.95) and threshold (from 0.74 to 0.95) models were moderate to high, respectively. Very high rank correlations (0.98 to 0.99) between EBV predicted for the same trait from linear and threshold model indicated that reranking of sires based on model used was minimal. In conclusion, despite low heritability, we found utilizable genetic variation in early lactation of heifers. Hence, genetic selection to improve genetic resistance to SCM in early lactation of heifers was deemed possible.

Lactoferrin and IgG levels in ovine milk throughout lactation: Correlation with milk quality parameters

The aim of this work has been to determine the levels of lactoferrin and IgG in ovine milk during the two first months of lactation and to compare with some milk quality parameters. Milk samples were collected from ewes of an Assaf flock. The levels of lactoferrin were determined by radial immunodiffusion. For this purpose, lactoferrin was isolated from ovine milk by chromatographic methods and polyclonal antibodies against it were obtained in rabbits. IgG levels were determined by a commercial sandwich ELISA. The mean concentration value of lactoferrin at the first day of lactation was 0.74 mg/ml and that for IgG of 9.25 mg/mL. The levels dropped sharply during the first three days of lactation and at the third week of lactation, the concentration in mature milk was of 0.07 mg/ml and 0.25 mg/ml for lactoferrin and IgG, respectively. There was no correlation between the levels of lactoferrin and IgG concentrations in ovine milk samples with those of total protein and bacterial count. A low positive correlation between lactoferrin and somatic cell counts (SCC) was found, which was more evident when samples were classified according to SCC levels. However, no significant differences were found with respect to IgG levels in milk samples, for high or low levels of SCC. Consequently, it seems that only high lactoferrin levels could be used as an indicator of subclinical mastitis.

Intramammary infections and somatic cell counts in meat and pelt producing ewes with clinically healthy udders

Mastitis in sheep is common and is important both from an economic and from a welfare point of view. It can be either clinical or subclinical, bacterial or, more seldom, lentiviral. There are no data on the national prevalence of subclinical mastitis (SCM) or intramammary infection (IMI), and causative pathogens, in ewes in Sweden.Diagnosis of SCM and/or IMI is based on inflammatory indicators, measured in milk and additional bacteriology. Somatic cell count (SCC) is the most widely used indicator of SCM and/or IMI and can be measured either directly or indirectly with e.g. California Mastitis Test (CMT). However, the use of SCC as an indicator for mastitis and/or IMI in ewes is not fully evaluated, especially not in meat- and pelt producing herds. The aim of this study was therefore to investigate the prevalence of IMI and bacterial panorama in ewes with clinically healthy udders, in meat- and pelt producing herds in Sweden. Moreover, we wanted to define a cut-off for SCC and CMT for detecting ewes with IMI that would be suitable for use under field conditions.Udder half milk samples (n=2134) were collected at weaning and lambing from meat- and pelt producing ewes (n=773), in 22 farms in Sweden. Only clinically healthy animals were included in the study. Milk samples were analysed for bacteriology and SCC and were given a CMT score.Intramammary infection was found in 30% of the ewes and in 14% of the udder half milk samples. No bacteria were found in 74% of the milk samples, and mixed flora was found in 12% of the samples. Among the 287 milk samples where IMI was identified, coagulase negative staphylococci (CoNS) were most prevalent (58%) followed by Staphylococcus (S.) aureus (9%) and Mannheimia (M.) haemolytica (6%). Among the 165 CoNS findings S. simulans was the most common (26%), followed by S. warneri, S. equorum, S. xylosus, S. haemolyticus and S. chromogenes. A CMT score of ≥3 and a SCC of ≥500,000 cells/ml at weaning or CMT score ≥3 and a SCC of ≥400,000 cells/ml after lambing gave the highest possible Se at the same time as the highest possible Sp for identifying udder halves with IMI and may be used for screening for IMI in Swedish meet and pelt producing herds.To conclude, one third of Swedish meat- and pelt producing ewes without clinical signs in the udder had IMI in one or both udder halves at weaning and or after lambing, and CoNS was the most common bacterial finding. A high SCC (≥400,000–500,000 cells/ml) was associated with IMI and a difference in having a high (CMT 3–5) or low (CMT 1–2) between udder halves can be used as an indicator of IMI under field conditions.

Use of milk amyloid A in the diagnosis of subclinical mastitis in dairy ewes

Subclinical mastitis (SM) is one of the most important diseases affecting dairy ewes worldwide, with negative impact on the animal health, farm income and public health. Animals with SM often remain untreated because the disease may not be revealed. Increase in somatic cell count (SCC) and positive bacteriology for mastitis pathogens in milk samples are indicative of SM but the evidence of only one of these alterations must suggest an uncertain SM (UM). UM is defined when positive bacteriological examination (Latent-SM) or SCC>500000cells/ml (non-specific-SM) are detected in milk. Nevertheless, SCC and bacteriological examination are expensive, time consuming and are not yet in use at the farm level in dairy ewes. Recently, a sensitive acute phase protein, amyloid A, displaying multiple isoforms in plasma and different body fluids including mammary secretion (milk amyloid A-MAA), has been investigated as a marker of mastitis in cows and, in a few studies, in sheep. The aim of this trial was to compare the concentration of MAA of single udder-halves in ewes with healthy udder-halves (HU-control group) and naturally occurring subclinical mastitis, both confirmed (SM group) and uncertain (UM groups: Latent-SM and non-specific-SM), for monitoring udder health. The reliability of a specific ELISA kit for the measurement of MAA was also tested. During a 3-month trial period, 153 udder halves were assigned to the experimental groups based on their health status: 25 with SM, 40 with UM (11 with latent-SM and 29 with non-specific-SM) and 88 HU. SCC and bacteriological analysis were performed to establish the control and subclinical mastitis groups. MAA concentrations in milk samples were measured using a specific commercially milk ELISA kit. The data were submitted to statistical analysis. Significant (P<0·05) differences among the groups SM, non-specific-SM and HU were detected with the SM having the highest level and HU the lowest. MAA concentration is affected by the udder health status and is a useful indicator of subclinical mastitis and increased SCC in sheep.

Associations of herd- and cow-level factors, cow lying behavior, and risk of elevated somatic cell count in free-stall housed lactating dairy cows

Recent evidence suggests that the risk of intramammary infection in dairy cows is related to lying patterns. The objectives of this study were to quantify the standing and lying behavior of dairy cows milked 3×/d, determine the cow- and herd-level factors associated with these behaviors, and relate these findings to the risk of an elevated somatic cell count (SCC). Five commercial free-stall dairy herds in Eastern Ontario, milking 3×/d, were enrolled in a longitudinal study. Forty Holstein-Friesian cows/herd were randomly selected as focal animals based on days in milk (<200d) and SCC (<100,000cells/mL). Farms were followed for 4, 5-week periods. Individual-cow SCC was recorded at the beginning of each period and end of the final period. Elevated SCC (eSCC) was used as an indicator of subclinical mastitis. A new incident eSCC was defined as an individual cow that started the period with a SCC <100,000cells/mL but whose next SCC exceeded 200,000cells/mL. Lying behavior was recorded 5d after each milk sampling using data loggers. For these 5d, individual milking times and feeding times were also recorded. On d1 of each recording period 2 trained observers scored focal cows for hygiene and lameness. Throughout the course of the study, cows averaged 11.2h/d of lying time, split into 8.6 lying bouts/d that were on average 84.6min in length. Later lactation cows had longer daily lying times that were split into fewer lying bouts of longer duration than cows earlier in lactation. Lame cows had longer daily lying times and lying bout durations than non-lame cows. Cows with greater milk yield had lower lying times than lower producing cows. Average post-milking standing time across the study herds was 103min. Manipulation of feed (feed delivery or push-up) by the stockperson, in the hour before milking or shortly thereafter, resulted in the longest post-milking standing times. Over the study period, 48 new eSCC were detected, resulting in a mean herd incidence rate of 0.91 eSCC/cow-year at risk for all study herds. A non-linear relationship between post-milking standing time and eSCC incidence was found; compared to those cows that lie down <90min after milking, cows that lie down for the first time >90min after milking had a lower risk of acquiring a new eSCC. The risk of experiencing an eSCC was also increased in multiparous cows, and in those cows with a higher SCC at the beginning of the study. These results indicate that management practices that promote post-milking standing time, such as the manipulation of feed delivery around milking times, should be encouraged to reduce the risk of cows experiencing new eSCC.

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.

Associations of dairy cow behavior, barn hygiene, cow hygiene, and risk of elevated somatic cell count

Poor dairy cow hygiene has been consistently associated with elevated somatic cell count (SCC) and the risk of subclinical mastitis. The objective of this study was to determine the associations between dairy cow standing and lying behavior, barn hygiene, cow hygiene, and the risk of experiencing elevated SCC. Lactating Holstein dairy cows (n=69; 86±51 DIM; parity: 2.0±1.2; means ± SD), kept in 1 of 2 groups, were monitored over a 4-mo period. Each group contained 61±1 (mean ± SD) cows over the study period; complete data were obtained from 37 and 32 animals within each respective group. Cows were housed in a sand-bedded, freestall barn with 2 symmetrical pens, each with a free cow traffic automatic milking system. To vary barn hygiene, in 4 consecutive 28-d periods, alley manure scrapers in each of the 2 pens were randomly assigned to frequencies of operation of 3, 6, 12, and 24 times per day. During the last 7dof each period, cow hygiene (upper leg/flank, lower legs, and udder; scale of 1 = very clean to 4 = very dirty) and stall hygiene (number of 0.15×0.15-m squares contaminated with manure in a 1.20×1.65-m grid) were recorded. Standing and lying behavior of the cows were collected during those days using data loggers. Individual-cow SCC was recorded at the beginning and end of each 28-d period. Elevated SCC was used as an indicator of subclinical mastitis; incidence of elevated SCC was defined as having a SCC >200,000 cells/mL at the end of each 28-d period, when SCC was <100,000 cells/mL at the beginning of the period. Less frequent scraping of the barn alleys was associated with cows having poorer hygiene. Poor udder hygiene was associated with poor stall hygiene. Longer lying duration was associated with poor hygiene of the upper legs/flank and udder. Greater premilking standing duration was associated with poor udder hygiene and decreased frequency of lying bouts was associated with poor hygiene of the lower legs. Higher milk yield was associated with poor hygiene of the udder and lower legs; multiparous cows had poorer hygiene of the upper legs/flank and udder. Over the study period, 24 new cases of elevated SCC were detected. No associations existed for the risk of experiencing an elevated SCC with alley scraping frequency or cow behavior patterns. However, increased odds of occurrence of elevated SCC were noted for cows of lower milk yield as well as for multiparous cows. In summary, these results show that cow hygiene is affected by the standing and lying behavior of cows and by the cleanliness of the cow’s environment. These findings emphasize the need for cows to be provided clean standing and lying environments. The results also show that frequent cleaning of barn alley floors will help improve cow hygiene.

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.

Assessment of Lactate Dehydrogenase, Alkaline Phosphatase and Aspartate Aminotransferase Activities in Cow’s Milk as an Indicator of Subclinical Mastitis

This study examined the activities of lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in the milk of lactating Holstein cows in association with subclinical mastitis (SCM). A total of 94 milk samples were collected from 58 lactating dairy cows representing stages of lactation from the second to the tenth week after calving. Those which were classified as positive by California mastitis test (CMT) were deemed to have subclinical mastitis. All the milk samples were skimmed by centrifugation at 10 000g at 0 degrees C and were used for enzyme activities estimations. The mean activities of LDH and ALP were higher in the milk from udders with SCM than in the milk from healthy udders (p < 0.05). There were no significant differences in AST values. The maximum agreement rates between the CMT results and LDH and ALP values were seen at thresholds of > 180 IU/L and > 40 IU/L respectively (kappa values 0.65 and 0.79, respectively). However, the sensitivity of the tests for identifying SCM at these thresholds was higher for ALP (96.4%) than for LDH (68.5%). In this study, LDH and ALP tests were standardized for cow's milk and results showed that only the ALP test was reliable in the early diagnosis of subclinical mastitis.

Breastmilk RNA viral load in HIV-infected South African women: effects of subclinical mastitis and infant feeding.

To investigate determinants of breastmilk RNA viral load among HIV-infected South African women, with particular attention to infant feeding mode and subclinical mastitis. Observational, longitudinal study. Information on current infant feeding practice and a spot milk sample from each breast were obtained from 145 HIV-infected lactating women at 1, 6 and 14 weeks postpartum. The sodium/potassium (Na+/K+) ratio in milk was taken as an indicator of subclinical mastitis. The association between milk RNA viral load and maternal and infant characteristics was investigated using uni- and multivariate models. Milk viral load was below the limit of detection of the HIV RNA assay (< 200 copies/ml) in 63/185 (34.1%), 73/193 (37.8%) and 68/160 (42.5%) of samples at 1, 6 and 14 weeks, respectively. Multivariate models predicted between 13 and 26% of variability in milk viral load in the first 14 weeks. Low blood CD4 cell count (< 200 x 10(6) cells/l) during pregnancy and raised milk Na+/K+ ratio were significantly associated with raised milk RNA viral load at all times, but there were no consistent associations between infant feeding mode and RNA viral load in milk. There was a non-significant trend for the six infants known to be infected postnatally, compared with the 88 infants who remained uninfected, to have been exposed to breastmilk of higher viral load at each time point. Breast milk HIV RNA viral load in the first 14 weeks of life varied; high levels were associated with subclinical mastitis and severe maternal immunosuppression. Multivariate models had limited predictive value for milk RNA viral load, illustrating the multiple contributors to viral load.