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Effect of stress on blood leucocyte and milk somatic cell counts in dairy cows.

Blood and milk samples from Holstein cows were examined for total blood leucocyte count, differential blood leucocyte count, milk quality test, and somatic cell count in milk while the cows were stressed by corticotropin injection, confinement in a heat-humidity chamber, or environmental-heat stress by exposure during the hot summer months of June through November in southern Arizona. All three stressing conditions resulted in a moderate blood leucocytosis. Modest increases in somatic cell counts of milk were associated with corticotropin injection and environmental-heat stress. Positive correlations were recorded between blood leucocytes and somatic cell counts of milk in mastitis-free cows injected with corticotropin and between percent blood neutrophils and somatic cell counts of milk in environmental-heat stressed cows with no evidence of current mastitis.

Adrenocorticotropic Hormone

Milk somatic cells in dairy cows kept on pasture or confined indoors during the summer.

During the summer of 1975 (from June 30th to July 28th) weekly milk samples were obtained from 49 cows kept on pasture during the daytime, and from 48 cows which were confined indoors in stanchion barns and fed grass twice a day. The cows on pasture were exposed to relatively moderate temperatures, which were somewhat lower than for the indoor group (Table I). In the cows kept on pasture, an increase in the milk somatic cell content (MSCC) was observed throughout the experimental period, whereas no corresponding increase was recorded in the indoor group (Table III). This increase in MSCC appeared to occur primarily in non-infected quarters (Table IV). The data from the present investigation seem to indicate that the seasonal increase in MSCC during the summer, which has been reported by several other investigators, cannot be directly related to environmental temperature per se.

Animal Feed

Influence of storage and preservation of milk samples on microscopic and Fossomatic somatic cell counts.

Milk somatic cell counting was carried out by Fossomatic and microscopically. In unpreserved milk samples Fossomatic counts increased by up to 100% during the first 24 hours after milking. During the next 24 hours the counts increased by 5% whereafter they remained stable until at least 80 hours after milking, when the samples were stored at 5 degress C. On microscopical counting using methylene blue for staining, the results were stable from shortly after milking. In counting, attention had to be paid to the fact that within the first 24 hours a certain part of the cells would stain but faintly. After preservation with potassium bichromate the Fossomatic counts increased more rapidly. Stable results were found 5--8 hours after preservation. The final level of the Fossomatic counts was found to be app. 10% higher in preserved than in unpreserved samples. A smaller increase was found by microscopic counting. The rise of the cell counts during the first 24 hours after milking is probably due to inadequate stainability of living cells with ethidium bromide, resulting in a certain part of them being recorded by the Fossomatic. During the first day the vitality of the cells diminishes whereby they become stainable and countable. This process in hastened by potassium bichromate treatment.

Animals

Relation between mastitis test score, mineral composition of milk, and blood electrolyte profiles in Holstein cows.

The relationship between mastitis, mineral composition of milk, and blood electrolyte profiles was investigated in 54 Holstein cows. Sodium potassium, calcium, magnesium, and chloride in blood and milk were compared under two indices of mastitis, a milk quality test and a milk somatic cell count. Milk from cows with evidence of udder infection had higher sodium and chloride and lower potassium than cows free of mastitis. Although there was a correlation between blood calcium and milk calcium and between milk calcium and milk somatic cell count, as well as differences in mean potassium and calcium in blood between mastitic and nonmastitic cows, there was no direct relationship between the indices of mastitis and electrolyte profiles of blood.

Animals

Effect of bovine neutrophil maturity on phagocytosis.

The ability of immature circulating neutrophils to phagocytose foreign particles was determined. One quarter of each of 4 cows, which were free of mastitis and pathogens and had milk somatic cell counts of less than 1 X 10(6) cells/ml, was infused with endotoxin of Escherichia coli (0.5 mg in 3 ml of saline solution) to induce a milk leukocytosis and a shift to the left in circulating leukocytes. Milk samples were taken from the infused and one uninfused quarter before and after infusion. Blood samples were collected at 9 hours after infusion, at 2-hour intervals from 13 to 21 hours, and at 36 hours after infusion. Phagocytosis was determined by incubating whole blood with yeast cells and determining the precentage of neutrophils phagocytosing and the number of yeast cells per neutrophil. Milk somatic cells increased from an average of less than 1.0 X 10(6) cells/ml before infusion to 37.0 X 10(6) cells/ml at the next milking (postinfusion hour 11). Circulating immature neutrophils increased from 7.6% before infusion to 56.6% at postinfusion hour 21. The percentage of neutrophils phagocytosing was 99 to 100 throughout the study. Number of yeast cells per neutrophil (mean phagocytosis) decreased from 6.5 before infusion was done to 5.2 after infusion. However, there was no correlation between percentage of immature neutrophils and mean phagocytosis. The decrease in yeast cells per neutrophil was attributed to the decrease in concentration of neutrophils and yeast cells in the in vitro procedure (r = 0.50). Apparently the immature neutrophils that appear after depletion of the mature form are as phagocytic as the mature neutrophils.

Animals

From buffalo to human: Klebsiella pneumoniae in high-somatic cell count milk as an overlooked link in the one health chain.

High somatic cell count (SCC) is a critical indicator of udder health and milk quality in buffalo milk production. However, in many low-income regions, SCC monitoring is often underemphasized, allowing a proportion of high-SCC buffalo milk to enter the food chain and potentially compromising food safety and public health. Klebsiella pneumoniae (K. pneumoniae) is a common zoonotic pathogen found in high-SCC milk, yet systematic investigations into the prevalence and characteristics in high-SCC buffalo milk remain limited. In this study, 23 K. pneumoniae strains were screened out from 460 bacterial isolates obtained from high-SCC buffalo milk samples from Guangxi, China, with an isolation rate of 5.0%. These isolates were comprehensively characterized using whole-genome sequencing and comparative genomic analyses. The results revealed that 78.26% (18/23) of the isolates shared high genomic similarity with the human reference strain ATCC 13883, and the ST37 clone exhibited a pronounced potential of cross-species transmission. All isolates harbored core adhesion factors and intrinsic resistance genes. Notably, several strains displayed high-risk features: strain 419 carried the K1 capsular serotype, strain 326 possessed a complete yersiniabactin synthesis gene cluster, and strain 320 exhibited a multidrug-resistant phenotype. Phenotypic assays further demonstrated a positive correlation between biofilm formation capacity and virulence in Galleria mellonella. Metabolic pathway enrichment analyses suggested that K. pneumoniae has undergone substantial adaptation to the nutrient-rich buffalo milk environment. Collectively, these findings confirm that raw high-SCC buffalo milk serves as a significant reservoir for high-risk zoonotic K. pneumoniae. While industrial thermal processing effectively eliminates viable pathogens, the resilient antimicrobial resistance determinants within these isolates pose a persistent risk of horizontal gene dissemination along the food chain, providing critical evidence for enhancing pre-processing milk quality regulations within a One Health framework.

Animals

Protein, casein, and noncasein protein percentages in milk with high somatic cell counts.

More than 1000 milk samples of individual cows with somatic cell counts ranging from 20,000 to 20,000,000 per ml were analyzed by amido black dye binding for total protein, whey protein, and casein. Ten classes of varying somatic cell counts were established. The mean total protein percentage for normal samples (counts less than 500,000/ml) was 3.2% and different from all the other protein class means, ranging from 3.4 to 3.9%. Total protein content increased with somatic cell count. There was no significant difference among any of the ten classes for casein values. Whey protein percentages for normal samples were different from the rest. Whey protein content increased with somatic cell count.

Animals

Cell volume to aid analysis and technique of somatic cell counts in milk.

In conjunction with a Coulter Counter, somatic cells in milk were sized by electronic analysis. Quarter milk from cows with mastitis had a cell volume peak with a modal cell volume of 102 mu3 while milk from healthy quarters had no peak. Bulk milks with a peak had higher cell counts than milks where there was no peak. Dimensions of peak from bulk milks were the same as from quarter milks. Modal cell volume and cell count varied after milks were fixed with varying concentrations and types of fixative. The concentration of fixative recommended by the International Dairy Federation was sufficient, but only marginally so, and it should be increased.

Animals

Enzymic methods for estimation of the somatic cell count in bovine milk. 1. Development of assay techniques and a study of their usefulness in evaluating the somatic cell content of milk.

Assay procedures were developed for a number of enzymes in milk which apparently originate from leucocytes. The enzymes studied were acid phosphatase, N-acetyl-beta-D-glucosaminidase, beta-glucuronidase, arylsulphatase, alpha-mannosidase, and catalase. Quarter-milk samples were analysed for enzyme activity and results compared with the electronic cell count and the Wisconsin Mastitis Test. All enzymes measured except acid phosphatase and alpha-mannosidase showed good correlation with the electronic cell count. Of the other 4 enzymes tested, beta-glucuronidase and arylsulphatase were unsuitable as diagnostic aids owing to the lengthy incubation periods required in their assay procedures. The assay of catalase, which involved the measurement of the initial rate of release of O2 using an O2 analyser apparatus, was rapid, sensitive and reasonably reliable, if fresh milk samples were used. The assay procedure for N-acetyl-beta-D-glucosaminidase was considered to be the most reliable, simple and rapid enzymic method for estimating the number of somatic cells in milk.

Acetylglucosaminidase

Long non-coding RNAs link DNA methylation to immune regulatory networks in bovine subclinical mastitis.

Long non-coding RNAs (lncRNAs) are emerging as important regulators of inflammatory and immune signaling, yet their contribution to bovine subclinical mastitis remains poorly defined. Here, we characterized the lncRNA expression landscape associated with disease in milk somatic cells of healthy and subclinical mastitic Vrindavani cattle. We identified 11,403 high-confidence lncRNAs, of which 104 were differentially expressed in subclinical mastitis (adjusted P&#x2009;<&#x2009;0.05; |log2FC| &#x2265; 1), with the vast majority upregulated in mastitic samples. Predicted cis- and trans-associated target analyses identified 637 non-redundant genes, and KEGG analysis identified 8 significantly enriched cis-associated pathways and 152 significantly enriched trans-associated pathways (adjusted P&#x2009;<&#x2009;0.05), predominantly enriched for immune and inflammation-related pathways. These findings prioritized a subset of mastitis-associated lncRNAs for subsequent methylation and interaction-network analyses. A subset of these lncRNAs further overlapped differentially methylated regions (DMRs), suggesting a potential association between lncRNA expression changes and DNA methylation alterations. Integration of lncRNA-miRNA and miRNA-mRNA interactions identified lncRNA-miRNA-mRNA interaction networks involving DMR-associated lncRNAs. Among the prioritized candidates, MSTRG.28878.1 showed overlap with a hypomethylated promoter-associated DMR, increased expression, and multiple connections within the predicted interaction network. Together, these findings identify candidate lncRNAs, methylation-associated loci, and predicted molecular interactions associated with bovine subclinical mastitis and provide a resource for future functional investigation of candidate non-coding RNA-associated mechanisms in disease.

Animals

Effects of parturition and lactation on blood and milk cell concentrations, corticosteroids, and neutrophil phagocytosis in the cow.

Blood differential leukocytes, erythrocytes, milk somatic cells, and neutrophil phagocytosis in 14 Holstein-Friesian cows were measured at 2-day intervals for 2 weeks before parturition, on the day of parturition, on postpartum days 1,2,5,10,15, and 30, and at monthly intervals throughout lactation. Circulating corticosteroids were measured in 6 cows on days -6, -4, -2, on the day of parturition, and on days +1, +2, +5, +10, +15, +30, and +60. Circulating neutrophils on day -2 on the day of parturition, on days 2, 10, and 15 averaged 3,363, 5,889, 3,085, 4,116, and 3,558/mm, respectively. The increase in neutrophils at parturition was accompanined by an increase in corticosteroids (2.7, 9.0, 11.1, 6.1, and 5.4 ng/ml of blood plasma on days -4 and -2, on the day of parturition, and on days +1 and +2, respectively). The phagocytic ability of circulating neutrophils was determined by incubating yeast cells with whole blood (30 minutes, 37 C) and microscopically counting yeast cells per neutrophil. Mean yeast cells per neutrophil decreased (P less than 0.01) during the 2nd week after parturition (7.4, 6.5, 6.6, and 7.2 on days +2, +10, +15, and +30, respectively). But this decrease was compensated for by an increase in circulating neutrophils during the same period.

Adrenal Cortex Hormones

Relationship of the estrous cycle to milk composition.

The relationship of the estrous cycle to milk composition was examined in 15 Holstein cows in first lactation which were approximately 45 days postpartum. Composite milk samples were collected twice daily and analyzed for progesterone, fat, protein, total solids, sodium, potassium, magnesium, calcium, and somatic cells. Milk progesterone was minimal (less than ng/ml) from day -1 to day 2 (day 0 = estrus) of the estrous cycle and then increased at a relatively constant rate through day 15. Milk yield and sodium and magnesium concentrations of the milk varied with days of the estrous cycle. Mean milk yield was highest and sodium and magnesium concentrations were lowest on day 1. However, sodium concentration of the milk was the only component that varied significantly during the 3 days centered on estrus (days -1, 0, and 1). No other milk component changed significantly during the estrous cycle. Although milk yield and composition varied during the estrous cycle, none of these components appeared to be a practical indicator of estrus.

Animals

Cell counts in bulked milk supplies from dairy farms of northern Nigeria.

Somatic cells in 596 milk samples collected from bulked supplies of 3 northern Nigeria dairy farms were counted by an electronic method following a standard method for the preparation of the milk samples. Mean somatic cell counts per ml indicating low level of infection were 158,597, 166,742 and 155,032 for the 3 farms, without any significant differences. Mean somatic cell counts per ml indicating herd mastitis averaged 354,768 +/- 66,348 and the pathogenic organisms isolated were Escherichia coli and Staphylocucus aureus. Counts useful for future regular monitoring of somatic cells in bulked milk supplies in northern Nigeria are presented.

Animals

Improved automated optical somatic cell counting method for raw milk: collaborative study.

A collaborative study was conducted to compare the improved automated optical somatic cell counting procedure (OSCC II) with the direct microscopic somatic cell counting method (DMSCC) in raw milk. Samples were prefixed with formaldehyde and introduced into an Auto-Analyzer system. Dilution, clarification, and cell counting were performed automatically. Five collaborators participated in the study; they analyzed 48 samples in duplicate, using 2 different sampling rates. The results were compared with DMSCC counts reported by 3 different analysts. Statistical results show that the standard deviation for the DMSCC method is 0.0825 and for the OSCC II, at a sampling rate of 80/hr, 0.0434. When results are compared, the OSCC II procedure is as accurate as, and is significantly more precise than, the DMSCC method. The faster sampling rate of the OSCC II at 120/hr has some effect on precision but little effect on accuracy. The method has been adopted as official first action.

Animals