Italian Journal of Animal Science

Cross-sectional study on the prevalence of contagious pathogens in bulk tank milk and their effects on somatic cell counts and milk yield


Source: Alfonso ZecconiORCID Icon,Francesca dell’Orco,Nicoletta Rizzi,Diego Vairani,Micaela Cipolla,Paolo PozziORCID Icon & Lucio Zanini

Highlights: First study on a large sample of Italian dairy herds on the prevalence of contagious pathogens in bulk tank milk samples. The prevalence value observed exceeded 50%.

First study estimating the prevalence of M. bovis in bulk tank milk in a large sample of Italian dairy herds, and the prevalence observed was 1.5%.

Prevalence of contagious pathogens has a significant influence on milk yield and SCC.

Bulk tank milk SCC confirmed to have a low accuracy to identify infected herds.

Keywords: Herd health, mastitis, somatic cell count, milk yield, antimicrobials
ABSTRACT: Data on the prevalence of major contagious pathogens in bulk tank milk (BTM) in Italy are generally not available. The availability of Real-Time PCR procedures (qPCR) to perform BTM analysis by represents an important step to define herd health status. Therefore, a cross-sectional epidemiological study was designed to assess the prevalence of contagious pathogens and Prototheca spp in BTM samples. The study was performed on 581 herds from four districts in the west Lombardy region of Italy. Additionally, the relationship between pathogens in BTM and SCC or milk yield; the presence of an association between four risk factors (district, herd size, average milk yield and SCC) with pathogens in BTM were assessed. The overall data showed that S. aureus was recovered in 42% of the herds, Str. agalactiae in 10%, Prototheca spp in 11% and M. bovis in 1.5% of the herds. The GLM model applied showed a significant influence of BTM results, district, herd size and their interactions on SCC and on milk yield variance. Particularly, S. aureus or Str. agalactiae have a significant effect on milk yield variability and, in a lesser extent, on SCC. The very high prevalence of contagious pathogens significantly affects milk characteristics and yield, thus affecting economic sustainability of the herds, and suggests the need to implement control programmes to decrease the prevalence of contagious pathogens, This will also allow to decrease the use of antimicrobials and to improve cow welfare.

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ARLA Newsletter The Golden Milk, 2019 Vol. 7

HOW TO REDUCE MEDICINE COST?


Mastitis therapy is often made very complicated, because it is started that you need to look at sensitivity testing of all bacteria you find.
Let me speak directly –Forget sensitivity testing except in cases of Klebsiella or Pseudomonas. all other mastitis causes just need treating for the bacteria detected and in most cases do not need antibiotics at all !!
This makes mastitis therapy very simple and cheap. Not only that, it also follows guidelines on prudent use antibiotics – using almost exclusively narrow spectrum antibiotics such as Pencilin G or 1st generation Cephalosporin.

The drug required depends on the bacteria detected. (So keep it simple). It is all about cure rates and the concentration of the drug at the infection site in the quarter.
You have different drugs of 3rd or 4th gen Cephalosporin and Fluoroquinolon that show big inhibition circles at sensitivity testing leading to speculation that you should use these drugs, but look closer and you will find that cure rates are not better, as the concentration of the drug at the infection site is toolow, and these drugs are often more expensive.

Reed more on page 2 and 3

Dairy Sci. 2019, 102

Association between teat skin colonization and intramammary infection with Staphylococcus aureus and Streptococcus agalactiae in herds with automatic milking systems.


Source: Line Svennesen1, Søren S. Nielsen2, Yasser S. Mahmmod3, Volker Krömker, Karl Pedersen4, Ilka C. Klaas2.

1 Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg C, Denmark. Electronic address: line.svennesen@sund.ku.dk.
2 Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg C, Denmark.
3 Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg C, Denmark; Infectious Diseases, Department of Animal Medicine, Faculty of Veterinary Medicine, Zagazig University, 44511-Zagazig, Sharkia Province, Egypt.
4 Department of Microbiology, University of Applied Sciences and Arts, 30453 Hannover, Germany.
5 National Veterinary Institute, Technical University of Denmark, 2800 Kongens Lyngby, Denmark
ABSTRACT: The objective of this study was to investigate the association between teat skin colonization and intramammary infection (IMI) with Staphylococcus aureus or Streptococcus agalactiae at the quarter level in herds with automatic milking systems. Milk and teat skin samples from 1,142 quarters were collected from 300 cows with somatic cell count >200,000 cells/mL from 8 herds positive for Strep. agalactiae. All milk and teat skin samples were cultured on calf blood agar and selective media. A subset of samples from 287 quarters was further analyzed using a PCR assay (Mastit4 PCR; DNA Diagnostic A/S, Risskov, Denmark). Bacterial culture detected Staph. aureus in 93 (8.1%) of the milk samples and 75 (6.6%) of the teat skin samples. Of these, 15 (1.3%) quarters were positive in both the teat skin and milk samples. Streptococcus agalactiae was cultured in 84 (7.4%) of the milk samples and 4 (0.35%) of the teat skin samples. Of these, 3 (0.26%) quarters were positive in both the teat skin and milk samples. The PCR detected Staph. aureus in 29 (10%) of the milk samples and 45 (16%) of the teat skin samples. Of these, 2 (0.7%) quarters were positive in both the teat skin and milk samples. Streptococcus agalactiae was detected in 40 (14%) of the milk samples and 51 (18%) of the teat skin samples. Of these, 16 (5.6%) quarters were positive in both the teat skin and milk samples.

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Preventive Veterinary Medicine

Accuracy of qPCR and bacterial culture for the diagnosis of bovine intramammary infections and teat skin colonisation with Streptococcus agalactiae and Staphylococcus aureus using Bayesian analysis


Source: Line Svennesen a, Yasser S.Mahmmod ab1, Nanna K.Skjølstrup a, Louise R.Mathiasen a, JørgenKatholm c, Karl Pedersen d, Ilka C.Klaas a2, Søren S.Nielsen a.

a Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, DK-1870 Frederiksberg C, Denmark
b Infectious Diseases, Department of Animal Medicine, Faculty of Veterinary Medicine, Zagazig University, 44511, Zagazig, Sharkia Province, Egypt
c DNA Diagnostic A/S, Voldbjergvej 14, 8240 Risskov, Denmark
d National Veterinary Institute, Technical University of Denmark, 2800, Kongens Lyngby, Denmark

ABSTRACT: Streptococcus agalactiae (Strep. agalactiae) and Staphylococcus aureus (Staph. aureus) are originally regarded as contagious mastitis pathogens, however, both pathogens have recently been isolated from extramammary and environmental sites, indicating that other sites than the udder might contribute to the spread of these pathogens potentially causing intramammary infections. Diagnostic tools to identify pathogens at extramammary sites are available but still needs to be validated. The objective of this cross-sectional field study was to estimate the diagnostic sensitivity (Se) and specificity (Sp) of the commercially available Mastit4 qPCR assay and bacterial culture (BC) in identifying Strep. agalactiae and Staph. aureus from milk and teat skin samples. We randomly selected 30–40 cows with high somatic cell counts from eight Danish Strep. agalactiae-positive dairy herds with automatic milking systems. Teat skin samples and aseptic milk samples were collected from right rear quarters (n = 287) for BC and PCR analysis. Se and Sp were estimated in a Bayesian latent class analysis. For milk samples, the Se and Sp of qPCR for Strep. agalactiae were estimated to 0.97 and 0.99, respectively, whereas the Se and Sp of BC were 0.41 and 1.00, respectively. The Se and Sp of qPCR for Staph. aureus were estimated to 0.95 and 0.99, respectively, whereas the Se and Sp of BC were 0.54 and 0.77, respectively. For teat skin samples,

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Journal of Dairy Science

Elimination of selected mastitis pathogens during the dry period


SOURCE: Anri A. E. Timonen 1, Jørgen Katholm1, Anders Petersen 1,Toomas Orro2, Kerli Mõtus 2 and Piret Kalmus 1.

1 Chair of Clinical Veterinary Medicine, Institute of Veterinary Medicine and Animal Science, Estonian University of Life Sciences, 51014 Tartu, Estonia
2 DNA Diagnostic A/S, Voldbjergvej 16, 8240 Risskov, Denmark.

ABSTRACT: We aimed to evaluate the elimination of 4 different mastitis pathogens, Streptococcus agalactiae, Myco-plasma bovis, Staphylococcus aureus, and Streptococcus uberis, from infected udder quarters during the dry period using quantitative PCR. The second purpose of this study was to evaluate the association between milk haptoglobin (Hp) concentration and the presence of udder pathogens (Strep. agalactiae, Staph. aureus, M. bovis, and Strep. uberis) in udder quarter milk samples before and after dry period. Aseptic udder quarter milk samples (n = 1,001) were collected from 133 dairy cows at dry off and at the first milking after calving from 1 large dairy herd. Bacterial DNA of Strep. agalactiae, Staph. aureus, Strep. uberis, and M. bovis in the udder quarter milk samples was identified with commercial quantitative PCR analysis Mastitis 4B (DNA Diag-nostic A/S, Risskov, Denmark). Milk Hp concentration (mg/L) was measured from udder quarter milk samples. The elimination rates during the dry period for M. bo-vis, Staph. aureus, Strep. agalactiae, and Strep. uberis were 86.7, 93.6, 96.2, and 100.0%, respectively. The new IMI rate was 3.0% for M. bovis, 2.9% for Staph. aureus, 2.4% for Strep. agalactiae, and 3.1% for Strep. uberis. The milk Hp concentration was significantly higher in udder quarter milk samples with blood and in samples positive for Strep. agalactiae at dry off and for Staph. aureus postcalving.

Read more in open access paper Journal of Dairy Science

JIA 2018, 06

Evaluation of a new qPCR test to identify the organisms causing high total bacterial count in bulk tank milk



SOURCE: Jørgen Katholm1, Lene Trier Olesen2, Anders Petersen1, Snorri Sigurdsson3

1 DNA Diagnostic A/S, Risskov 8240, Denmark
2 ARLA Foods amba, Viby 8260, Denmark
3 SEGES, Aarhus N 8200, Denmark
ABSTRACT: Milk quality in bulk tank milk (BTM) is measured by flow cytometry technology as total bacterial count (TBC) and somatic cell count (SCC). To investigate SCC problems, culture or PCR can be used to identify mastitis causing bacteria, e.g., Mastit 4, a commercially available qPCR test. TBC in BTM can be investigated further using culture-based methods such as standard plate count, laboratory pasteurization count, coliform count, and spore counts. To our knowledge, no qPCR addressing the bacteria involved in TBC has been commercially introduced. The aim of this study is to evaluate a recently introduced 3-h qPCR test, TBC 4. The TBC 4 qPCR detects four target groups, Pseudomonas, Streptococci, Enterobacteriacea/ Enterococcus, and Bacillus/Clostridia. These target groups relate to problems on the farm such as cooling, mastitis, environment, and silage. We will continue with new research to compare the TBC 4 qPCR test with traditional culture. For this study, BTM samples from different TBC intervals were selected based on BactoCount results found at routine payment investigation at Eurofins laboratory (Vejen, Denmark). These samples were analyzed using TBC 4 qPCR assay within 24 h. In total, 346 BTM samples were divided into six different intervals of colony forming units (CFU).

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Preventive Veterinary Medicine

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



SOURCE: Ingrid H. Holmøy, Nils Toft, Hannah J. Jørgensen, Tormod Mørk, Liv Sølverød,. and Ane Nødtvedt


ABSTRACT: Streptococcus agalactiae (S. agalactiae) has re-emerged as a mastitis pathogen among Norwegian dairy cows. The Norwegian cattle health services recommend that infected herds implement measures to eradicate S. agalactiae, this includes a screening of milk samples from all lactating cows. The performance of the qPCR-test currently in use for this purpose has not been evaluated under field conditions. The objective of this study was to estimate the sensitivity and specificity of the real-time qPCR assay in use in Norway (Mastitis 4 qPCR, DNA Diagnostics A/S, Risskov, Denmark) and compare it to conventional acteriological culturing for detection of S. agalactiae in milk samples. Because none of these tests are considered a perfect reference test, the evaluation was performed using latent class models in a Bayesian analysis. Aseptically collected cow-composite milk samples from 578 cows belonging to 6 herds were cultured and tested by qPCR. While 37 (6.4%) samples were positive for S. agalactiae by bacteriological culture, 66 (11.4%) samples were positive by qPCR.

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Journal of Dairy Science

Within-herd prevalence thresholds for herd-level detection of mastitis pathogens using multiplex real-time PCR in bulk tank milk samples



SOURCE: J. B. Soltau, E. Einax, K. Klengel, J. Katholm, K. Failing, A. Wehrend, and K. Donat
ABSTRACT: The objective of the study was to assess the value of quantitative multiplex real-time PCR examination of bulk tank milk samples for bovine mastitis pathogens as a tool for herd level diagnosis. Using a logistic regression model, this study is aimed at calculating the threshold level of the apparent within-herd prevalence as determined by quarter milk sample cultivation of all lactating cows, thus allowing the detection of a herd positive for a specific pathogen within certain probability levels. A total of 6,335 quarter milk samples were collected and cultured from 1,615 cows on 51 farms in Germany. Bulk tank milk samples were taken from each farm and tested by bacterial culture as well as the commercial PCR assay Mastit 4A (DNA Diagnostic A/S, Risskov, Denmark) identifying Staphylococcus aureus, Streptococcus dysgalactiae, Streptococcus agalactiae, and Streptococcus uberis.

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Journal of Dairy Science

Within-herd prevalence of intramammary infection caused by Mycoplasma bovis and associations between cow udder health, milk yield, and composition

Published online: June 7, 2017

SOURCE: Anri A.E. Timonen, Jørgen Katholm, Anders Petersen, Kerli Mõtus, Piret Kalmus
ABSTRACT: Subclinical mastitis is one of the major health problems in dairy herds due to decreased milk production and reduced milk quality. The aim of this study was to examine the within-herd prevalence of subclinical intramammary infection caused by Mycoplasma bovis and to evaluate associations between M. bovis and cow daily milk yield, udder health, and milk composition. Individual cow composite milk samples (n = 522) were collected from all lactating dairy cows in 1 Estonian dairy farm in November 2014. Daily milk yield, days in milk, and parity were recorded. Collected milk samples were analyzed for somatic cell count, milk protein, fat, and urea content.

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ICAR The Global Standard For Livestock Data

Proceedings of the 40th ICAR Biennial Session held in Puerto Varas, Chile, 24-28 October 2016


SOURCE: J. Katholm and A. Pedersen, DNA Diagnostic
The use of Real-time PCR tests to identify mastitis pathogenes are growing, as they are faster and more sensitive than conventional bacteriological culturing, especially for Mycoplasma detection. Since 2014 a quantitative real-time PCR test kit (Mastit 4, DNA diagnostic) has been commercially available. the objective of this study was to investigate the correlation between Ct values of the Mastit 4 PCR test kit for and bacterial colony forming units (CFU) in fresh milk samples for the 11 bacteria that can be detected by the Mastit 4 BDF, CFU, PCR test kit.

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NMC Annual Meeting Proceedings (2017)

Providing Routine PCR Testing as Part of Official Milk Recordings Options


SOURCE: Angela Dawn Coburn, Agsource Cooperative Service, Verona, Wisconsin USA
Introduction: Obtaining Somatic Cell Count (SCC) value from routine milk sample analysis is, historically, a primary reason U.S. dairy producers participate in official milk recordings program. With improves management of environmental conditions and general cow health, SCC levels have shown a steady decline over the past 4 years. Among Agsource herds, weighted average SCC for Holstein have dropped from 263,000 in 2012 to 227,000 in 2016. The percent of cows with SCC over 200,000 SCC has dropped from 24,3 % to 20,4 %.

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Jena at IDF Nantes Mastitis Conference (2016)

Jena at IDF Nantes Mastitis conference


SOURCE: J. B. Soltau, E. einax, K. Klengel, K. Donat, Animal Health Service, Thuringian animal Diseases Fund, Jena, Thuringia Germany. J. Katholm, DNA diagnostic, Risskov Denmark. K. Failing, Unit for Biomathermatics and Data Processing, Justus-liebig University, Giessen, Hesse Germany. A. Wehrend, Clinic of Obstetrics, gynaecology and Andrology for small and large Animals, Justus-Liebig University, Giessen, Hesse Germany.
Introduction: The aim of the study was to analyse the value of a polymerase chain reaction (PCR) examination of bulk tank samples for bovine mastitis pathogens as a tool to estimate the within herd prevalenceof these pathogens.
A total of 6,335 quarter milk samples were collected from 1,615 cows farms in Germany. Additionally, two bulk tank milk samples were collected from each farm. The quarter milk samples were cultivated in the laboratory of Animal Disease Fund (Jena, Thuringia, Germany). The bulk tank milk samples were tested by the commercial PCR assay Mastit 4 (DNA Diagnostic, Denmark) that identified Staphylococcus aureus, Streptococcus dysgalactiae, Streptococcus agalactiae and Streptocuccus uberis.

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NMC Annual Meeting Proceedings (2016)

Test characteristics of the qPCR test Mastit 4 to identify Major Pathogenes in spiked and originally infected Milk samples.


SOURCE: Torben W. Bennedsgaard, Line Svennesen, Ilka C. Klaas Aarhus University, Foulum Denmak and University of Copenhagen Denmark
Introduction: Real-time PCR tests are increasingly used to identify mastitis pathogenes as they are faster and more sensitive than conventional bacteriogical culturing. Recently a new commercially availeble quantitative real-time PCR test kit (Mastit 4, DNA Diagnostic) was introduced in Denmak and is mainly used to test cows prior to dry -off for subclinical intramammary infections or in herd screenings for eradication of Streptocuccus agalactiae. The objective of our study was to investigate the association between ct values of the Mastit 4 PCR test kit for and colony forming units in fresh milk samples from quaters without clinical mastitis for Streptocuccus agalactiae, Staphylocuccus aureus, Streptocuccus uberis and Streptococcus dysgalactiae.

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NMC Annual Metting Proceedings (2015)

Bulk Tank milk Surveillance For Contagios Mastitis Pathogenes: Comparison Of Two Commercial Real-Time PCR Test Kits


SOURCE: Erik Rattenborg, Carl O. Paulrud, Søren K. S. Jensen and Jørgen Katholm
Knowledge center for agriculture, Cattle, Aarhus N, Denmak
Eurofins, Vejen, Denmark
DNA diagnostic, Risskov, Denmark
As part of the Danish Surveillance program for Streptococcus agalactia. bulk tank milk (BTM) samples from all dairy herds are tested annually. since 2009 the bulk tank samples has been analyzed using real-time PCR PathoProof Mastitis assay (Thermo fisher Scientific, Vantaa, Fiinland) In 2009 and 2010 the analyses were carried out with the complete 12 Kit and in 2011-2013 with the complete 16 kit. Indseptember 2014 the real-time PCR Mastit 4 B (DNA Diagnostic, risskov, Denmark) was made commercially availeble. the objective of this study was to compare the PathoProof Mastitis Complete 16 kit with the Mastit 4b kit regarding the detection of four species of bacteria tested by both test (Staphylococcus aureus, Streptococcus agalactiae, Streptococcus uberis and Mycoplasma bovis) the comparison reveals essential knowledge tha may be used in designing the future surveillance program.

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Veterinary Microbiology, 242, March 2020, 108608

Dynamics of the within-herd prevalence of Mycoplasma bovis intramammary infection in endemically infected dairy herds.


Anri Aino Elisa Timonen, Tiina Autio, Tarja Pohjanvirta, Liidia Häkkinen, Jørgen Katholm, Anders Petersen, Kerli Mõtusa, Piret Kalmus.


A B S T R A C T

We aimed to identify the dynamics of the within-herd prevalence of Mycoplasma (M.) bovis intramammary infection (IMI) in four dairy herds, estimate prevalence of M. bovis in colostrum and clinical mastitis cases and compare M. bovis strains from calves’ respiratory and cow clinical mastitis samples.

Within a six-month study period, cow composite milk samples (CMS) were collected three times during routine milk recording, first milking colostrum samples from all calving cows and udder quarter milk samples from clinical mastitis cases. Calf respiratory samples were collected from calves with respiratory disease. Pooled milk samples were analysed for M. bovis with the Mastitis 4B polymerase chain reaction (PCR) test kit (DNA Diagnostic A/S).

Prevalence estimates were calculated with Bayesian framework in R statistical programme. cg-MLST was used for M. bovis genotyping. In Herd I and II first testing M. bovis IMI within-herd prevalence (95 % credibility interval (CI)) was 4.7 % (2.9; 6.8) and 3.4 % (2.3; 4.6), changing to 1.0 % (0.1; 1.7) and 0.8 % (0.1; 1.4) in Herd I and 0.4 % (0.0;0.7) in Herd II at the next samplings. In Herd III and IV first testing M. bovis IMI within-herd prevalence was 12.3% (9.7; 15.2) and 7.8 % (6.2; 9.5), changing to 4.6 % (3.0; 6.4) and 3.2 % (1.9; 4.8) in Herd III and to 2.8 % (1.9;3.8) and 4.9 % (3.6; 6.4) in Herd IV at the next samplings. The estimated prevalence of M. bovis in colostrum ranged between 1.7 % (0.2; 2.8) and 4.7 % (2.7; 7.1) and in clinical mastitis cases between 3.7 % (1.7; 6.4) and 11.0 % (7.5; 15.2) in the study herds. M. bovis strains isolated from cows and calves clustered within herds indicating possible transmission of M. bovis between dairy cows and calves. Prevalence of M. bovis in colostrum and clinical mastitis cases as well as the within-herd prevalence of M. bovis IMI was low in endemically infected dairy herds.

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Who is DNA Diagnostic A/S

DNA Diagnostic A/S is a privately owned Danish biotech company established in 1992 developing and producing several PCR and qPCR test kits.

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DNA Diagnostic A/S
Voldbjergvej 14
DK-8240 Risskov
Denmark

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Phone: +45 87 32 30 50
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