Retrospective evaluation of udder recovery of cows with subclinical mastitis following treatment with acoustic pulse technology (APT) on commercial dairy farms and its economic impact.

Retrospective evaluation of udder recovery following treatment of the inflamed quarter with acoustic pulse technology (APT) of cows with subclinical mastitis was done on 4 Israeli commercial dairy farms. Here, we evaluated the APT treatment as a tool to manage subclinical mastitis and its economic consequences in commercial farms. Recovery of the infected glands following APT treatment was compared to the customary no-treatment (NT) for cows with subclinical mastitis. Over 2 years, 467 cows with subclinical mastitis were identified. Subclinical mastitis was defined by elevated somatic cell count (SCC; >1 × 106 cells/mL) in the monthly test-day milk sample; 222 cows were treated with APT and 245 cows were not treated and served as control. Differences between treatment groups in culling, milk quality, milk yield and bacterial elimination were analyzed. After treatment, cure from bacteria was calculated only for cows with pre-isolated bacteria. The percentage of sampled cows determined as cured (no bacterial finding) in the NT group was 32.7% (35/107) (30.9% Gram negative; 32.4% Gram positive) and in the APT-treated group, 83.9% (42/55) (89.4% Gram negative; 80.6% Gram positive). Culling rate due to mastitis was significantly lower (>90%) in the APT-treated vs. NT group. Recovery was 66.0% in the APT group compared to 11.5% in the NT group at 90 d post-treatment. Average milk volume per cow in the APT-treated group was 16.1% higher compared to NT cows. Based on the study, savings incurred by using APT to treat only subclinical cows per 100-cow herd can total $15,106/y, or $309 per treated subclinically infected cow.

Management of high cows-share-contribution of SCC to the bulk milk tank by acoustic pulse technology (APT).

A cow with mastitis has a high somatic cell count (SCC) in its milk. Cow-share-contribution of somatic cells to the bulk milk tank (BMTSCC) refers to the relative addition made by each cow's milk to the bulk tank's SCC. Since bulk milk is graded and priced according to the BMTSCC, high-yielding cows with mastitis are the main contributors to penalizations in milk price. The benefits of acoustic pulse technology (APT) application to tissues are well documented, including its anti-inflammatory effect and restoration of tissue function by triggering natural healing processes. An APT-based device was developed specifically for treating mastitis in dairy cows. It enables rapid and deep penetration of the acoustic pulses over a large area of the udder in a single session. A study was performed on six farms with a total of 3,900 cows. One unit of cow-share-contribution equaled the addition of 1,000 cells to each mL of the bulk milk volume above the mean BMTSCC. A total of 206 cows were selected: 103 were treated with APT and 103 served as controls. All of the cows contributed over 1.5 units to the BMTSCC at the time of treatment. Seventy-five days after APT treatment, 2 of the 103 treated cows (1.9%) were culled, compared to 19 (18.5%) of the 103 control cows, as well as infected quarter dry-off in 5 others (4.85%). Overall success was defined as a decrease of >75% in cow-share-contribution from treatment time in two of the three monthly milk recordings following treatment. Results indicated 57.3% success for the APT-treated cows vs. 14.6% for the untreated control groups. Highest share-contribution provide an additional tool for the farmer's decision of how to control BMTSCC. Because the cow-share-contribution value is relative to herd size and BMTSCC, this study included a similar number of cows, with similar SCC and milk yield from each of the six herds.

Physiological response of mammary glands to Escherichia coli infection:A conflict between glucose need for milk production and immune response.

The mammary immune and physiological responses to distinct mammary-pathogenic E. coli (MPEC) strains were studied. One gland in each of ten cows were challenged intra-mammary and milk composition (lactose, fat, total protein, casein), biochemical (glucose, glucose-6-phosphate (Glu6P), oxalate, malate, lactate, pyruvate and citrate, malate and lactate dehydrogenases, lactate dehydrogenase (LDH), nitrite, lactic peroxidase, catalase, albumin, lactoferrin, immunoglobulin) and clotting parameters were followed for 35 days post-challenge. Challenge lead to clinical acute mastitis, with peak bacterial counts in milk at 16-24 h post-challenge. Biochemical and clotting parameters in milk reported were partially in accord with lipopolysaccharide-induced mastitis, but increased Glu6P and LDH activity and prolonged lactate dehydrogenase and Glu6P/Glu alterations were found. Some alterations measured in milk resolved within days after challenge, while others endured for above one month, regardless of bacterial clearance, and some reflected physiological responses to mastitis such as the balance between aerobic and anaerobic metabolism (citrate to lactate ratios). The results suggest that E. coli mastitis can be divided into two stages: an acute, clinical phase, as an immediate response to bacterial infection in the mammary gland, and a chronic phase, independent of bacteria clearance, in response to tissue damage caused during the acute phase.

Correlation between Milk Bacteriology, Cytology and Mammary Tissue Histology in Cows: Cure from the Pathogen or Recovery from the Inflammation.

The aim of the current study was to verify the existence of a significant correlation between bacterial isolation (or not) and mammary gland inflammation, using traditional bacterial culturing and PCR, milk leucocytes distributions, and tissue histology. Twenty-two cows were tested at the level of the individual gland for bacteriological culture and real-time PCR (RT-PCR), milk composition, somatic cells count (SCC), and cell differentiation. Post-slaughter samples of teat-ends and mammary tissues were tested for histology and bacteriology by RT-PCR. The 88 glands were assigned to either outcome: 1. Healthy-no inflammation and no bacterial finding (NBF) (n = 33); 2. Inflammation and NBF (n = 26); 3. Inflammation and intra-mammary infection (n = 22) with different bacteria. Bacteriology of milk samples and that of the RT-PCR showed 91.4% agreement. In the lobule's tissues of healthy glands, ~50% were milk producers and the other glands had dry areas with increased fat globules with a low number of leukocytes. In contrast, ~75% of the infected glands were identified as inflamed, but with no isolation of bacteria. Infiltration of mononuclear cells and neutrophils into the connective tissue was observed but not in the lobule's lumen. In summary, the study confirms that not every mastitis/inflammation is also an infection.

Milk quality and milk transformation parameters from infected mammary glands depends on the infecting bacteria species.

The current study measured the influence of milk of subclinically infected glands by different bacteria species on the cow's milk. The effects of bacterial infection or inflammation on gland milk yield were related to the bacteria species that caused the infection. The volume of milk of the inflamed gland from the cow's milk yield was significantly lower (P<0.001) for the glands previously infected by Escherichia coli (PIEc) and those infected with Streptococcus dysgalactiae. Coagulation properties, rennet clotting time (RCT) and curd firmness (CF) also depended on the bacteria causing the infection. RCT values of all the inflamed glands were significantly longer (P<0.001) and CF values were significantly lower than that of the healthy ones. Moreover, in the whole milk, CF was also significantly lower and not proportional to the volume of the milk from the inflamed gland of the cow's milk. Calculation of the predicted 40% dry matter curd weight (PCW) on the cow level, including the healthy and inflamed glands or the healthy glands alone, showed that for 10 of 13 PIEc cows, the presence of the affected gland's milk in the whole cow milk resulted in a lower PCW value. Likewise, 7 of 20 cows infected by S. dysgalactiae had negative delta values. Unlike the latter bacteria, PCW from milk of glands infected with CNS increased, although in a lower magnitude than in the healthy glands. No correlation was found between logSCC in the whole cow milk (healthy and inflamed glands) and PCW.

The Interrelationships between Lactose Intolerance and the Modern Dairy Industry: Global Perspectives in Evolutional and Historical Backgrounds.

Humans learned to exploit ruminants as a source of milk about 10,000 years ago. Since then, the use of domesticated ruminants as a source of milk and dairy products has expanded until today when the dairy industry has become one of the largest sectors in the modern food industry, including the spread at the present time to countries such as China and Japan. This review analyzes the reasons for this expansion and flourishing. As reviewed in detail, milk has numerous nutritional advantages, most important being almost an irreplaceable source of dietary calcium, hence justifying the effort required to increase its consumption. On the other hand, widespread lactose intolerance among the adult population is a considerable drawback to dairy-based foods consumption. Over the centuries, three factors allowed humans to overcome limitations imposed by lactose intolerance: (i) mutations, which occurred in particular populations, most notably in the north European Celtic societies and African nomads, in which carriers of the lactose intolerance gene converted from being lactose intolerant to lactose tolerant; (ii) the ability to develop low-lactose products such as cheese and yogurt; and (iii) colon microbiome adaptation, which allow lactose intolerant individuals to overcome its intolerance. However, in a few examples in the last decade, modern dairy products, such as the popular and widespread bio-cultured yogurts, were suspected to be unsuitable for lactose intolerant peoples. In addition, the use of lactose and milk-derived products containing lactose in non-dairy products has become widespread. For these reasons, it is concluded that it might be important and helpful to label food that may contain lactose because such information will allow lactose intolerant groups to control lactose intake within the physiological limitations of ~12 g per a single meal.

Influence of intramammary infection of a single gland in dairy cows on the cow's milk quality.

Intramammary infection (IMI), comprises a group of costly diseases affecting dairy animals worldwide. Many dairy parlours are equipped with on-line computerised data acquisition systems designed to detect IMI. However, the data collected is related to the cow level, therefore the contribution of infected glands to the recorded parameters may be over estimated. The present study aimed at evaluating the influence of single gland IMI by different bacteria species on the cow's overall milk quality. A total of 130 cows were tested 239 times; 79 cows were tested once and the others were examined 2-8 times. All of the analysed data refer to the number of tests performed, taking into account the repeated testing of the same cows. Of the cows tested ~50% were free of infection in all 4 glands and the others were infected in one gland with different coagulase negative staphylococci (CNS), Streptococcus dysgalactiae, or were post infected with Escherichia coli (PIEc), i.e., free of bacterial infection at the time of sampling but 1-2 months after clinical infection by E. coli. Overall, infection with bacteria had significant effects on somatic cell count (SCC) and lactose concentration. Examining each bacterium reveals that the major influence on those parameters was the sharp decrease in lactose in the PIEc and curd firmness in PIEc and Strep. Individual gland milk production decreased ~20% in Strep. dysgalactiae- and ~50% in PIEc-infected glands with respect to glands with no bacterial findings. Significant differences were found in lactose, SCC, rennet clotting time and curd firmness in the milk of infected glands and among those, these parameters were significantly higher in Strep. dysgalactiae and PIEc than in CNS infected cows. The current results using quarter-milking reinforces the importance of accurate IMI detection in relation to economic and welfare factors, and moreover, emphasises the need for technical sensing and constant reporting to the farmer about changes in the milk quality of every animal.

Milk metabolites as indicators of mammary gland functions and milk quality.

The assumption, that metabolites derived from the activity of the mammary gland epithelial cells reflect changes in milk secretion and its coagulation properties, was tested in dairy cows. The experiment included cows with uninfected udders and cows with one of the glands infected by different bacteria specie. Analysis were carried at the cow level (including all four glands), or at the gland level. High and significant correlations among the concentrations of lactose, glucose, glucose-6-posphate, milk related respiratory index (the ratio between the concentrations of citrate/lactate+malate in milk) and milk-derived glycolytic index (the ratio between glucose-6-phosphate and glucose in milk) and milk clotting parameters were found. The physiological basis for these relations and their ability to predict the deterioration in milk quality in subclinically infected glands and in glands previously clinically infected with Escherichia coli are discussed.

Subclinical mastitis in goats is associated with upregulation of nitric oxide-derived oxidative stress that causes reduction of milk antioxidative properties and impairment of its quality.

The aim of this study was to verify the existence of a nitric oxide (NO) cycle in goat milk and to study how changes in it affect milk composition during subclinical mastitis. Fifteen lactating dairy goats in which one udder-half was free from bacterial infection and the contra-lateral one was naturally infected with various species of coagulase-negative staphylococci were used. In comparison to uninfected glands, subclinical mastitis was associated with a decrease in milk yield, lactose concentration, and curd yield and an increase in nitrite and nitrate concentrations and with measurements reflecting increased formation of NO-derived free-radical nitrogen dioxide. The occurrence of NO cycling in goat milk was largely confirmed. The increase in the NO-derived stress during subclinical infection was not associated with significant increase in oxidatively modified substances, 3-nitrotyrosine, and carbonyls on proteins, but with increased levels of peroxides on fat. However, the relatively modest nitrosative stress in subclinically infected glands was associated with significant reduction in total antioxidant capacity and vitamin C levels in milk. We concluded that subclinical mastitis in goats caused by coagulase-negative staphylococci imposes negative changes in milk yield, milk quality for cheese production, and negatively affects the nutritional value of milk as food. Thus, subclinical mastitis in goats should be considered as a serious economic burden both by farmers and by the dairy industry.

Early mammary gland metabolic and immune responses during natural-like and forceful drying-off in high-yielding dairy cows.

The present work compared metabolic and immune responses in genetically high-producing cows that produced a low amount of milk before expected involution and in cows with the same genetic potential that produced copious amounts of milk before their scheduled drying-off. Ten multiparous lactating Israeli Holstein cows producing approximately 10,500 L in the current lactation, without bacterial infection and scheduled for drying-off approximately 60 d before their expected parturition, were studied. Five of the cows that exhibited a sharp, spontaneous reduction in milk yield at the end of their lactation and produced less than ~14L/d were defined as cows approaching natural involution (ANI), and 5 cows that produced between 25 and 35 L/d were defined as cows approaching forced involution (AFI). Three days before scheduled drying-off, milking was stopped and milk samples were collected from each quarter. After milking cessation, only modest swelling was observed in the udders of the ANI cows. In the ANI cows, lactose and fat concentrations decreased and the fat:lactose concentration ratio indicated that on d 1 and 2 fat concentrations decreased faster than lactose concentration, whereas on d 3, the rate of reduction was about the same for lactose and fat. In contrast, in AFI cows, fat concentrations increased on d 1 and the fat:lactose ratio indicated that changes in fat secretion were minor compared with those of lactose secretion. Rennet clotting time of milk after drying-off in the ANI cows increased, whereas curd firmness decreased rapidly, such that mammary secretions did not coagulate on d 3. In the AFI cows, such significant changes were observed only on d 3. The inflammatory response increased in both groups, but at each stage the increase was greater in ANI cows than in AFI cows. On d 1, the increase in leukocyte numbers in the ANI cows was made up of mononuclear cells (i.e., T lymphocytes and macrophages). In contrast, in the AFI cows, we observed a marked increase in leukocyte numbers, mainly in the form of polymorphonuclear cells. Our data indicate that the abrupt mammary involution induced in AFI cows provoked signs of distress, which were associated with neutrophilia in milk. In contrast, in the ANI cows, cessation of milking occurred without evidence of engorgement of the udder. Physiological differences in ANI and AFI cows are distinct and are reflected in the differences in the leukocyte populations in milk.

Tissue-type plasminogen activator and plasminogen embedded in casein rule its degradation under physiological situations: manipulation with casein hydrolysate.

The aims of this study were to test the assumption that tissue-type plasminogen activator (t-PA) and plasminogen (PG) are closely associated with the casein micelle and form a functional complex that rules casein degradation. This assumption was essentially verified for bovine milk under conditions wherein the plasmin system was activated by treatment with casein hydrolysate. It was also shown that urokinase-type PA (u-PA), the second type of plasminogen activator present in milk, was not involved in casein degradation. In agreement with previous studies, we show that treatment with casein hydrolysate precipitously reduced mammary secretion, disrupted the tight junction integrity (increase in Na+ and decrease in K+ concentrations), induced hydrolysis of casein, and activated various elements of the innate and acquired immune system. In the present study, we have identified t-PA as the principal PA, which is responsible for the conversion of PG to plasmin. It was found that t-PA and plasminogen are present in freshly secreted milk (less than 10 min from its secretion), suggesting that they are secreted as a complex by the mammary gland epithelial cells. Further research is needed to provide the direct evidence to verify this concept.

Real-time evaluation of milk quality as reflected by clotting parameters of individual cow's milk during the milking session, between day-to-day and during lactation.

Real-time analysis of milk coagulation properties as performed by the AfiLab™ milk spectrometer introduces new opportunities for the dairy industry. The study evaluated the performance of the AfiLab™ in a milking parlor of a commercial farm to provide real-time analysis of milk-clotting parameters -Afi-CF for cheese manufacture and determine its repeatability in time for individual cows. The AfiLab™ in a parlor, equipped with two parallel milk lines, enables to divert the milk on-line into two bulk milk tanks (A and B). Three commercial dairy herds of 220 to 320 Israeli Holstein cows producing ∼11 500 l during 305 days were selected for the study. The Afi-CF repeatability during time was found significant (P < 0.001) for cows. The statistic model succeeded in explaining 83.5% of the variance between Afi-CF and cows, and no significant variance was found between the mean weekly repeated recordings. Days in milk and log somatic cell count (SCC) had no significant effect. Fat, protein and lactose significantly affected Afi-CF and the empirical van Slyke equation. Real-time simulations were performed for different cutoff levels of coagulation properties where the milk of high Afi-CF cutoff value was channeled to tank A and the lower into tank B. The simulations showed that milk coagulation properties of an individual cow are not uniform, as most cows contributed milk to both tanks. Proportions of the individual cow's milk in each tank depended on the selected Afi-CF cutoff. The assessment of the major causative factors of a cow producing low-quality milk for cheese production was evaluated for the group that produced the low 10% quality milk. The largest number of cows in those groups at the three farms was found to be cows with post-intramammary infection with Escherichia coli and subclinical infections with streptococci or coagulase-negative staphylococci (∼30%), although the SCC of these cows was not significantly different. Early time in lactation together with high milk yield >50 l/day, and late in lactation together with low milk yield<15 l/day and estrous (0 to 5 days) were also important influencing factors for low-quality milk. However, ∼50% of the tested variables did not explain any of the factors responsible for the cow producing milk in the low - 10% Afi-CF.

Effects of intra-mammary bacterial infection with coagulase negative staphylococci and stage of lactation on shedding of epithelial cells and infiltration of leukocytes into milk: comparison among cows, goats and sheep.

The effects of mammary gland bacterial infection and stage of lactation on leukocyte infiltration into the mammary gland were compared among cows, goats and sheep. Animals were at two stages of lactation: mid or late. In mid-lactation animals, bacterial-free glands and coagulase negative Staphylococcus (CNS)-infected glands were compared. In late lactation only uninfected glands were studied. Of mid-lactation bacteria-free animals, goats had the highest number of leukocytes and % polymorphonuclears (PMNs), whereas sheep had the lowest and leukocytes number in cows were intermediate between sheep and goats. Based on %PMN, two cell clusters were found in sheep, which overlapped with the parallel cell clusters of cows and goats, but with a slightly higher number of leukocytes in each cell cluster. At late lactation, goats had higher values for %PMN and leukocyte numbers in comparison to cows, which had a similar cellular profile to sheep. The cellular immune response to CNS infection was similar for the three animal species, although the number of cells was different, while the basal cell level at mid-lactation and especially at the end of lactation was species specific.

Hydrogen peroxide-dependent conversion of nitrite to nitrate as a crucial feature of bovine milk catalase.

The enzyme catalase is well-known to catalyze the disintegration of hydrogen peroxide to water and oxygen; however, this study shows that its main function in bovine milk is oxidation of nitrite to nitrate. This process depends on hydrogen peroxide, of which the main source appears to be hydrogen peroxide formation that is coupled to the conversion of purines--xanthine in the present study--to uric acid by milk xanthine oxidase. However, additional secondary sources of hydrogen peroxide appear to be important during the relatively long storage of milk in the gland cistern. This paper demonstrates that the oxidation of nitrite to nitrate is necessary to prevent accumulation of free radicals and oxidative products during storage of milk in the gland and during the unavoidable delay between milking and pasteurization in dairy plants. Recommendations for minimizing the deterioration in milk quality during commercial storage are presented.

Aetiology of intramammary infection and its effect on milk composition in goat flocks.

Although there has been little study of the origin of intramammary infection (IMI) in goats, a common view is that most bacterial infection in goats occurs during milking. In the present study, the dynamics of occurrence of udder infection during and between lactations in three Anglo-Nubian goat farms in Israel was monitored. Coagulase-negative staphylococci were the predominant bacteria in the IMIs. We found that about 15% of the yearling does were already infected with bacteria when they joined the flock, whereas about 8% of the goats that dried-off returned with new IMIs. Moreover, virtually none of the goats acquired infection during lactation. Thus, our study showed that the aetiology of IMI in goats is very similar to that in dairy cows. A preventive treatment during the dry period should, therefore, be considered as an effective means of reducing the current rate of bacterial infections in goats.

Effect of subclinical intramammary infection on somatic cell counts, NAGase activity and gross composition of goats' milk.

The study was aimed at identifying the pathogens causing subclinical udder infections in representative Israeli dairy goat herds and determining their effect on milk quality. Five hundred goats in ten flocks of various breeds and crossbreeds were surveyed. Of the 500 goats, 13.4% were in their first lactation, 36.4% were in their second lactation and 50.2% were in their third or higher lactation. Percentages of udder halves with subclinical intramammary infection in the flocks ranged from 35 to 71%. The effect of the bacteriological infection on somatic cells count (SCC) was significant (P<0.001). Various species of coagulase-negative staphylococci (CNS), mainly Staphylococcus caprae and Staphylococcus epidermidis, were the main pathogens in infected udder halves. Lactation number did not significantly influence either infection rate of udder halves or SCC, although the percentage of udder halves with no bacteriological findings was higher at the first lactation than at the third lactation. Milk composition (fat, protein and lactose) varied among flocks, with lower mean total protein in uninfected halves than in infected ones and higher lactose in uninfected than infected halves.

Thermal inactivation of Escherichia coli in camel milk.

Camels subsist and produce milk in desert pastures not utilized by other domesticated herbivores. Developing the camel milk industry can improve the economy of desert inhabitants. To comply with sanitary ordinances, camel milk is pasteurized by procedures specified for bovine milk. It is widely accepted that milk composition might affect bacterial thermal death time (TDT). Camel and bovine milks markedly differ in their chemical composition, yet data regarding TDT values of bacteria in camel milk is missing. As a first step toward developing specific heat treatments appropriate for camel milk, TDT curves of Escherichia coli in artificially contaminated camel and cow milks have been compared. Heating the milks to temperatures ranging from 58 to 65 degrees C yields similar thermal death curves and derived D- and z-values. These findings suggest that, in this temperature range, E. coli might behave similarly in bovine and camel milk. Additional TDT studies of various pathogenic species in camel milk are required before establishing pasteurization conditions of camel milk.