Capture immunoassay for the diagnosis of bovine mastitis using a monoclonal antibody to polymorphonuclear granulocytes

1992 ◽  
Vol 59 (2) ◽  
pp. 123-133 ◽  
Author(s):  
Catherine A. O'Sullivan ◽  
Patrick J. Joyce ◽  
Teresa Sloan ◽  
Alan G. Shattock
Keyword(s):  
Monoclonal Antibody ◽  
Somatic Cell ◽  
Bovine Milk ◽  
Bovine Mastitis ◽  
Cell Counting ◽  
Enzyme Linked Immunosorbent Assay ◽  
Assay Sensitivity ◽  
Milk Samples ◽  
Cell Counts ◽  
Direct Elisa

SummaryA direct capture enzyme-linked immunosorbent assay (ELISA) was developed to measure elevated polymorphonuclear granulocyte (PMN) antigens using horseradish peroxidase (EC 1.11.1.7) conjugated rabbit polyclonal anti-PMN antisera and a monoclonal antibody specific for PMN cells. Optical densities obtained in the ELISA were used to predict the cell counts of milk samples. Predicted counts were not significantly different from actual somatic cell counts (SCC). In a total of 156 bovine milk samples the correlation coefficient between somatic cell counting, taking > 500000 cells/ml as being indicative of mastitis, and the assay was 0·94, yielding an assay sensitivity of 95·2% and a specificity of 97·3%. In further trials the ELISA could detect elevated PMN antigens in milk with SCC as low as 100000 cells/ml. The results indicate that the monoclonal antibody based direct ELISA has excellent potential in the detection and determination of bovine mastitis.

scholarly journals Bacterial identification, somatic cell count, antimicrobial profile and toxigenic Staphylococcus strains search from mastitic cow milk samples on small farms properties

2019 ◽  
Vol 39 (9) ◽  
pp. 715-722
Author(s):  
Ubirajara L. Lavor ◽  
Felipe F. Guimarães ◽  
Anelise Salina ◽  
Mateus S.R. Mioni ◽  
Helio Langoni
Keyword(s):  
Public Health ◽  
Somatic Cell ◽  
Negative Impact ◽  
Bovine Milk ◽  
Bovine Mastitis ◽  
High Sensitivity ◽  
Cow Milk ◽  
Small Farms ◽  
Milk Samples ◽  
Cell Counts

ABSTRACT: Bovine mastitis has a negative impact on milk production and can pose risks to public health. The present study aimed to evaluate the quality of bovine milk from small farms in the Botucatu/SP region. Somatic cell counts (SCC), identification of pathogens involved in mastitis, and sensitivity antimicrobial profile of staphylococci isolated were performed. The presence of enterotoxin encoding genes in isolates of staphylococci obtained from milk was investigated. Milk samples from individual mammary quarters of cows were submitted to the California mastitis test (CMT) and SCC. Of the 239 dairy cows from 21 dairy herds evaluated (mean = 11.4 animals/property), two cows (0.8%) presented clinical mastitis and 86 (35.9%) subclinical mastitis. Bacterial culture was performed in 177 quarter milk samples. Staphylococci were identified in 55 (31.1%), corynebacteria in 45 (25.4%), streptococci in 25 (14.1%) and coliforms in four (2.3%) milk samples. Average SCC from culture-positive samples was 1598x103 cells/mL, in case of staphylococci was 1362x103 cells/ml, streptococci was 2857x103 cells/mL, corynebacteria was 976x103 cells/mL and in the cases of coliforms 1161x103 cells/mL were obtained. Staphylococci showed a high sensitivity (>95%) to cephalothin, cotrimoxazole, enrofloxacin, and gentamicin, with a 41.2% resistance to penicillin and 11.8% to oxacillin. Both coagulase positive (CPS) and negative staphylococci (CNS) carried genes encoding enterotoxins in 21.6% of the first group and 41.9% in the second. The sea gene was the most detected 45.8% (n=24) between them, followed by seb with 29.2% and sec with 25.0%. The sed gene was not identified. We highlight the potential risk to public health in the possibility of strains of Staphylococcus spp. enterotoxin-producing genes that can cause staphylococcal food poisoning.

Streptococcus spp. from bulk-tank milk and milking machine teatcups on small ruminant farms, and factors potentially associated with their isolation

2020 ◽  
Vol 87 (3) ◽  
pp. 277-281
Author(s):  
Dimitris C. Chatzopoulos ◽  
Daphne T. Lianou ◽  
Charalambia K. Michael ◽  
Dimitris A. Gougoulis ◽  
Vasia S. Mavrogianni ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Small Ruminant ◽  
Milk Composition ◽  
Cell Counting ◽  
Bulk Tank Milk ◽  
Milk Samples ◽  
Streptococcus Uberis ◽  
Cell Counts ◽  
Bulk Tank ◽  
Milking Machine

AbstractThe objectives of this work were (a) to determine the presence of streptococci in samples from small ruminant dairy farms (bulk-tank milk and, where possible, teatcup swabs), (b) to investigate the potential adverse effects of streptococci on milk quality and (c) to investigate the importance of some husbandry factors for the isolation of streptococci. Bulk-tank milk samples and teatcups swab samples were examined bacteriologically for the presence of streptococci. Somatic cell counting and milk composition measurements were also performed. The husbandry factors present in each farm were assessed for potential associations with the isolation of streptococci. Streptococci were isolated from milk samples from 31.4% of sheep and 17.4% of goat farms and from 4.8% of sheep and 5.9% of goat teatcups. Streptococci were isolated more frequently from the upper part than the lower part of teatcups: 5.0% vs. 1.9%. Most isolates (57.9%) were identified as Streptococcus uberis. Most isolates (68.4%) were slime-producing; slime-production was more frequent among isolates from teatcups (83.3%) than from bulk-tank milk (55.0%). Somatic cell counts and milk composition did not differ between farms in which streptococci were or were not isolated. Machine-milking was associated with the isolation of streptococci from bulk-tank milk samples. The initial stage of the milking period (first two months) was found to be associated with the isolation of streptococci from milking machine teatcups in sheep farms only.

Detection of Zearalenone By Tandem Immunoaffinity-Enzyme-Linked Immunosorbent Assay and Its Application to Milk

1990 ◽  
Vol 53 (7) ◽  
pp. 577-580 ◽  
Author(s):  
JUAN I. AZCONA ◽  
MOHAMED M. ABOUZIED ◽  
JAMES J. PESTKA
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Monoclonal Antibody ◽  
Initial Sample ◽  
Average Recovery ◽  
Milk Samples ◽  
Coefficients Of Variation ◽  
Direct Elisa ◽  
Commercial Milk

A hybridoma-based method utilizing tandem affinity chromatography and enzyme-linked immunosorbent assay (ELISA) was devised to detect zearalenone. A zearalenone specific monoclonal antibody was attached to Sepharose for initial sample clean-up. Zearalenone was eluted with methanol and then quantified by competitive direct ELISA. Average ELISA recoveries from the column for water spiked with zearalenone at levels of 1, 5, 10, 25, and 50 ng/ml were 107, 86, 95, 95, and 92%, respectively, with a mean recovery of 95%. Mean interwell and interassay coefficients of variation were 9.7 and 8.9%, respectively. Average recovery by the method from milk spiked with zearalenone at levels of 1, 5, 10, 25, and 50 ng/ml was 187, 113, 107, 110, and 112%, respectively, with a mean recovery of 126%. Mean interwell and interassay coefficients of variation were 14.5 and 9.1%, respectively. Zearalenone was not detectable in 12 commercial milk samples assayed by the tandem method.

Preparation and Use of Somatic Cell Count Samples (SCCS) for Comparison of Milk Somatic Cell Counting Methods

1987 ◽  
Vol 50 (2) ◽  
pp. 132-135 ◽  
Author(s):  
T. J. LINTNER ◽  
A. L. LANGE ◽  
C. W. HEALD ◽  
R. J. EBERHART
Keyword(s):  
Somatic Cell ◽  
Cell Count ◽  
Somatic Cell Count ◽  
Bovine Milk ◽  
Coulter Counter ◽  
Cell Counting ◽  
Cell Content ◽  
Cell Counts ◽  
Counting Methods ◽  
Before And After

Somatic cell count samples (SCCS) for use in comparison of milk somatic cell counting methods were prepared from the cell sediment deposited in a creamery milk separator. Bovine milk somatic cells were resuspended from the sediment, and serial cell dilutions were prepared in bronopol-preserved milk diluent. Over a 1-year period, sets of SCCS were prepared each month and sent to milk-testing laboratories in the U.S.A., Canada and Europe, and counted by the methods in use at those Laboratories: (a) direct microscopic somatic cell count (DMSCC), (b) Fossomatic counter and (c) Coulter counter. Cell counts were normalized to eliminate the effect of month to month variation in the cell content of the SCCS. Counts obtained by the three methods were similar, although Coulter counter results tended to be lower, and significantly lower (P< 0.05) in SCCS with cell counts greater than 700,000 cells/ml than those counts by the other two methods. The effect of shipping on SCCS stability was assessed for SCCS samples sent to and returned from other laboratories, and counted by the Fossomatic method on their return. Counts were similar before and after shipping, except that results for SCCS with cell counts greater than 1,000,000 cells/ml were significantly higher (P<0.05) after their return.

Collaborative Study to Determine Effects of Milk Composition on Somatic Cell Counts

1990 ◽  
Vol 53 (1) ◽  
pp. 67-71 ◽  
Author(s):  
J. S. HOGAN ◽  
K. LARRY SMITH ◽  
D. A. TODHUNTER ◽  
P. S. SCHOENBERGER
Keyword(s):  
Somatic Cell ◽  
Protein Content ◽  
Collaborative Study ◽  
Fat Content ◽  
Cell Counting ◽  
Bulk Tank Milk ◽  
Somatic Cell Counts ◽  
Milk Samples ◽  
Cell Counts ◽  
Bulk Tank

Quarter, composite, and bulk tank milk samples were analyzed in a three laboratory collaborative study to determine the relationship of milk fat and protein content with milk somatic cell counts. Milk somatic cell counts were determined by two Coulter counters, a Fossomatic counter, and by direct microscopic somatic cell counting. In general, variability among somatic cell counts measured by different procedures was not related to protein or fat content of milk. The greatest percentage of variation between counts that could be explained by fat content of milk was 20.2% between a Coulter and direct microscopic somatic cell counts. The greatest percentage of variation between counts that could be explained by protein content of samples was 12.9% between a Coulter and Fossomatic counts. Breed of cow from which samples were collected also had little influence on differences among milk somatic cell counts. Differences among milk somatic cell counts due to counting methods did vary among quarter, composite, and bulk tank milk samples.

scholarly journals Relationships of Alizarin Test Scores with Somatic Cell Count in Bovine Milk

2020 ◽  
Vol 8 (10) ◽  
pp. 2089-2091
Author(s):  
Baqdaulet Sadikov ◽  
Savaş Atasever
Keyword(s):  
Somatic Cell ◽  
Cell Count ◽  
Correlation Coefficient ◽  
Test Scores ◽  
Somatic Cell Count ◽  
Bovine Milk ◽  
Cell Counting ◽  
Milk Samples ◽  
Ph Values

The aim of this study was to reveal the association of alizarin test scores (ATS) with somatic cell scores (SCC) in bovine milk. The milk samples (n=60) were collected from three milk selling points in Samsun province of Turkey in two test days (TD). To obtain ATS, change of colour in milk after adding dye solution were used (ATS 1: red-brown/normal, ATS 2: yellow/acidic, and ATS 3: violet/alkali). To test for reliability of ATS by acidity, pH values of the samples were measured with a standard pH meter. In SCC analyses, direct microscopic cell counting was applied. Of examined parameters, only SCC was differed by TD, and both pH and SCC were differed by the location. According to ATP groups, pH and SCC means were also changed. Finally, estimated weak (r=0.247) correlation coefficient between ATS and SCC pointed out that ATS data are not suitable to detect quality degree of bovine milk samples.

Frequency of individual udder quarters with elevated CMT scores in cows' milk samples with low somatic cell counts

2004 ◽  
Vol 155 (7) ◽  
pp. 213-213 ◽  
Author(s):  
I. Berglund ◽  
G. Pettersson ◽  
K. Svennersten‐Sjaunja ◽  
K. Östensson
Keyword(s):  
Somatic Cell ◽  
Somatic Cell Counts ◽  
Milk Samples ◽  
Cell Counts

scholarly journals Extensive Countrywide Field Investigation of Somatic Cell Counts and Total Bacterial Counts in Bulk-Tank Raw Milk in Sheep Flocks in Greece

Foods ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 268
Author(s):  
Daphne T. Lianou ◽  
Charalambia K. Michael ◽  
Natalia G. C. Vasileiou ◽  
Efthymia Petinaki ◽  
Peter J. Cripps ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Geometric Mean ◽  
Cell Counting ◽  
Lactation Period ◽  
Mechanical Ventilators ◽  
Bulk Tank Milk ◽  
Bacterial Counts ◽  
Somatic Cell Counts ◽  
Cell Counts ◽  
Bulk Tank

Objectives were to investigate somatic cell counts (SCC) and total bacterial counts (TBC) in the raw bulk-tank milk of sheep flocks in Greece, to study factors potentially influencing increased SCC and TBC in the bulk-tank milk of sheep and to evaluate possible associations of SCC and TBC with milk content. Throughout Greece, 325 dairy sheep flocks were visited for collection of milk sampling for somatic cell counting, microbiological examination and composition measurement. Geometric mean SCC were 0.488 × 106 cells mL−1; geometric mean TBC were 398 × 103 cfu mL−1; 228 staphylococcal isolates were recovered form 206 flocks (63.4%). Multivariable analyses revealed annual incidence risk of clinical mastitis, age of the farmer and month into lactation period (among 53 variables) to be significant for SCC > 1.0 × 106 cells mL−1 and month into lactation period at sampling and availability of mechanical ventilators (among 58 variables) to be significant for TBC > 1500 × 103 cfu mL−1. Negative correlation of SCC with fat, total protein and lactose and positive correlation of SCC with added water were found. With SCC > 1.0 × 106 cells mL−1, significant reduction of protein content (2%) was observed, whilst in flocks with SCC > 1.5 × 106 cells mL−1, significantly lower annual milk production per ewe (42.9%) was recorded.

scholarly journals Rapid Detection of Mycoplasma bovis, Staphylococcus aureus and Streptococcus agalactiae in Cattle Bulk Tank Milk in Cyprus and Relations with Somatic Cell Counts

Pathogens ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 841
Author(s):  
Maria Liapi ◽  
George Botsaris ◽  
Costas Arsenoglou ◽  
Nikolas Markantonis ◽  
Christodoulos Michael ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Somatic Cell ◽  
Real Time ◽  
Streptococcus Agalactiae ◽  
Mycoplasma Bovis ◽  
Bulk Tank Milk ◽  
Somatic Cell Counts ◽  
Milk Samples ◽  
Cell Counts ◽  
Bulk Tank

One hundred and seventy-seven (177) bulk tank milk samples were analyzed with a commercially available real-time polymerase chain reaction kit and 11 (6.21%), 41 (23.16%), and 58 (32.77%) tested positive for Mycoplasma bovis, Staphylococcus aureus, and Streptococcus agalactiae, respectively. Statistical analysis revealed a significant relationship between the presence of S. aureus and S. agalactiae. Enumeration of somatic cells was performed in the same samples by flow cytometry. The somatic cell counts were found higher in S. aureus and S. agalactiae positive samples. No association was found between M. bovis presence and somatic cells counts. Low internal assay control Ct values were found to be related with high somatic cell counts. Noticeably, this is the first report for the presence of M. bovis in Cyprus. Therefore, its presence was confirmed by bulk tank milk culture, conventional PCR, and next generation sequencing. Furthermore, M. bovis was typed with multilocus sequencing typing and was allocated to sequence type 29 (ST 29). Real-time PCR in bulk tank milk samples is a useful tool to detect mammary infections, especially for neglected pathogens such as M. bovis.

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