Microbiota in milk from healthy and mastitis cows varies greatly in diversity, species richness and composition, as revealed by PacBio sequencing

AbstractMastitis is the most economically important disease of dairy cows. This study used PacBio single-molecule real-time sequencing technology to sequence the full-length of the l6S rRNA from the microbiota in 27 milk samples (18 from mastitis and 9 from healthy cows; the cows were at different stages of lactation). We observed that healthy or late stage milk microbiota had significantly higher microbial diversity and richness. The community composition of the microbiota from different groups also varied greatly. In milk from healthy cows the microbiota was predominantly comprised of Lactococcus lactis, Acinetobacter johnsonii and Bacteroides dorei, while from mastitis cows it was predominantly comprised of Bacillus cereus, Clostridium cadaveris and Streptococcus suis. The prevalence of La. lactis and B. cereus in milk from healthy and mastitis cows was confirmed by digital droplets PCR. Differences in milk microbiota composition could suggest an important role for these microbes in protecting the host from mastitis. Based on the milk microbiota profiles, the Udder Health Index was constructed to predict the risk of bovine mastitis. Application of this predictive model could aid early identification and prevention of mastitis in dairy cows, though the model requires further optimisation using a larger dataset.

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PacBio sequencing revealed variation in the microbiota diversity, species richness and composition between milk collected from healthy and mastitis cows

Microbiology ◽

10.1099/mic.0.000968 ◽

2021 ◽

Vol 167 (7) ◽

Author(s):

Teng Ma ◽

Lingling Shen ◽

Qiannan Wen ◽

Ruirui Lv ◽

Qiangchuan Hou ◽

...

Keyword(s):

Single Molecule ◽

Type Species ◽

Cow Milk ◽

Sequencing Technology ◽

Pacbio Sequencing ◽

Content Type ◽

Milk Samples ◽

Link Type ◽

Microbiota Diversity ◽

Important Disease

Mastitis is the economically most important disease of dairy cows. This study used PacBio single-molecule real-time sequencing technology to sequence the full-length 16S rRNAs from 27 milk samples (18 from mastitis and nine from healthy cows; the cows were at different stages of lactation). We observed that healthy or late stage milk microbiota had significantly higher microbial diversity and richness. The community composition of the microbiota of different groups also varied greatly. The healthy cow milk microbiota was predominantly comprised of Lactococcus lactis , Acinetobacter johnsonii , and Bacteroides dorei , while the milk from mastitis cows was predominantly comprised of Bacillus cereus . The prevalence of L. lactis and B. cereus in the milk samples was confirmed by digital droplets PCR. Differences in the milk microbiota diversity and composition could suggest an important role for some these microbes in protecting the host from mastitis while others associated with mastitis. The results of our research serve as useful references for designing strategies to prevent and treat mastitis.

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Mycoplasmal mastitis in dairy cows in the Moghan region of Ardabil State, Iran : short communication

Journal of the South African Veterinary Association ◽

10.4102/jsava.v77i4.382 ◽

2006 ◽

Vol 77 (4) ◽

Cited By ~ 5

Author(s):

C. Ghazaei

Keyword(s):

Dairy Cows ◽

Short Communication ◽

Bovine Mastitis ◽

Subclinical Mastitis ◽

Clinical Mastitis ◽

Mycoplasma Bovis ◽

Bulk Tank Milk ◽

Milk Samples ◽

The World ◽

Bulk Tank

Mycoplasmas are an important and economically significant cause of mastitis in dairy cows in various parts of the world. The organisms are highly contagious, with the main reservoir of infection originating from cows with subclinical mastitis. In 1998 the 1st cases of bovine mastitis due to Mycoplasma bovis were diagnosed in Ardabil State, Iran. An investigation was carried out with the aim of establishing the extent of mycoplasma infections in dairy cows in Ardabil State. Milk samples obtained from 80 cows with clinical mastitis were cultured in the laboratory for the presence of mycoplasmas. Similarly, 48 bulk-tank milk samples were examined for the presence of mycoplasmas. A modified Hayflick broth was used to isolate the mycoplasmas and an immunoperoxidase test used for the species identification of the isolates. Mycoplasma bovis was isolated from 39 (48.75 %) of the clinical mastitis samples and from 48 of the bulk-tank milk samples tested. This indicated that mycoplasma udder infections were more prevalent in dairy cows in Ardabil State than previously thought.

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Yeasts as a causative agent of bovine mastitis in Greece

Journal of the Hellenic Veterinary Medical Society ◽

10.12681/jhvms.15257 ◽

2018 ◽

Vol 54 (2) ◽

pp. 105 ◽

Cited By ~ 3

Author(s):

E. BOURTZI-HATZOPOULOU (Ε. ΜΠΟΥΡΤΖΗ-ΧΑΤΖΟΠΟΥΛΟΥ) ◽

A. ZDRAGAS (Α. ΖΔΡΑΓΚΑΣ) ◽

E. PETRIDOU (Ε. ΠΕΤΡΙΔΟΥ) ◽

G. FILIOUSIS (Γ. ΦΙΛΙΟΥΣΗΣ)

Keyword(s):

Mammary Gland ◽

Dairy Cows ◽

Bovine Mastitis ◽

Economic Loss ◽

Geotrichum Candidum ◽

Clinical Mastitis ◽

Antibacterial Treatment ◽

Northern Greece ◽

Milk Samples ◽

Mastitic Milk

The aim of this study was to isolate fungi from mastitic milk of dairy cows and to identify fungal microorganisms involved in bovine mastitis. A total of 608 milk samples from clinical mastitis quarters from 580 animals in Northern Greece were collected, during the years 1997-2001. Antibacterial treatment was administrated to 50% of the sampled animals. Forty two (42) fungi were isolated and identified using mycological media and methods. From the 42 fungi isolates, 38 were yeasts and 4 moulds. The yeasts isolated were classified into the genera Candida, Geotrichum, Rhodotorula. From the thirty four (34) Candida species, 14 were identified as G tropicalis and 6 as G krusei. Furthermore, 4 isolates were classified as G pseudotropicalis, 4 as G albicans, 3 as G parapsilosis and 3 as G rugosa. Geotrichum candidum and Rhodotorula spp. were represented with 2 isolates. The isolated moulds were classified into the genus Aspergillus. Fungi were isolated in pure culture from 38 milk samples and in mixed culturewith bacteria from 4. In 510 samples only bacteria were cultured, while in 56 samples no growth of microorganism was observed. The results of the present study indicate that a level of 6.9% of mycotic mastitis is significant and yeasts are apparently implicated in mammary gland pathology causing economic loss.

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Tryptophan, Kynurenine and Kynurenic Acid Concentrations in Milk and Serum of Dairy Cows with Prototheca Mastitis

Animals ◽

10.3390/ani11123608 ◽

2021 ◽

Vol 11 (12) ◽

pp. 3608

Author(s):

Mariola Bochniarz ◽

Tomasz Piech ◽

Tomasz Kocki ◽

Mateusz Iskra ◽

Henryk Krukowski ◽

...

Keyword(s):

Dairy Cows ◽

Cell Count ◽

Somatic Cell Count ◽

Kynurenic Acid ◽

Bovine Mastitis ◽

Dairy Herds ◽

Serum Samples ◽

Milk Samples ◽

California Mastitis Test ◽

The Difference

The aim of this work was to investigate serum and milk levels of tryptophan (TRP), kynurenine (KYN), and kynurenic acid (KYNA), as well as the activity of indoleamine 2,3-dioxygenase (IDO) in cows with mastitis due to Prototheca algae. The study was prompted by previous research showing a link between the KYN pathway of TRP metabolism and bovine mastitis of bacterial etiology. The study was carried out over a 2-year period (2018–2019) and included quarter milk and serum samples collected from six dairy herds in Poland. The samples were obtained from healthy cows and cows with Prototheca mastitis of either clinical and subclinical manifestation, as determined upon direct measurement of the somatic cell count or indirectly by performing a California Mastitis Test on suspected quarters. Both TRP and KYN concentrations were significantly lower in milk of mastitic cows compared to healthy animals (0.8 vs. 8.72 µM, p = 0.001; 0.07 vs. 0.32 µM, p = 0.001, respectively). The difference in TRP and KYN concentrations in the sera of the two animal groups was much less pronounced (25.55 vs. 27.57 µM, 3.03 vs. 3.56 nM, respectively). The concentration of KYNA was almost at the same level in milk (1.73 vs. 1.70 nM) and in serum (80.47 vs. 75.48 nM) of both mastitic and healthy cows. The data showed that the level of TRP and its metabolites in serum was conspicuously higher compared to milk in all cows under the study. The activity of IDO was significantly higher in milk of cows with Prototheca mastitis compared to healthy animals (71.4 vs. 40.86, p < 0.05), while in serum it was pretty much the same (135.94 vs. 124.98, p > 0.05). The IDO activity differed significantly between serum and milk both for mastitic (135.94 vs. 71.4, p < 0.05) and healthy cows (124.98 vs. 40.86, p < 0.001). In conclusion, low values of TRP and KYN concentrations or elevated IDO activity in milk samples might be used as markers of mastitis due to infectious causes, including Prototheca spp.

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Changes of physicochemical indicators during mastitis and the effects of milk ejection on their sensitivity

Journal of Dairy Research ◽

10.1017/s0022029904000366 ◽

2004 ◽

Vol 71 (3) ◽

pp. 316-321 ◽

Cited By ~ 26

Author(s):

Rupert M Bruckmaier ◽

Daniel Weiss ◽

Martin Wiedemann ◽

Susanne Schmitz ◽

Georg Wendl

Keyword(s):

Escherichia Coli ◽

Dairy Cows ◽

Milk Ejection ◽

Udder Health ◽

Somatic Cell Counts ◽

Milk Samples ◽

Cell Counts ◽

Experimental Inflammation ◽

First Contact ◽

The Relationship

We examined the relationship between physicochemical indicators and somatic cells in the milk of dairy cows during experimentally induced mastitis and their significance as indicators for use in controlling udder health. We were concerned particularly with the effect of alveolar milk ejection on the sensitivity of these indicators. In Expt 1, Escherichia coli lipopolysaccharide (Esch. coli LPS) was injected into the left rear quarter to induce an inflammatory reaction in one quarter in each of six cows. The contralateral control quarter was injected with a solution of NaCl (9 g/l). Nine milk samples were taken from both quarters until 60 h after injection. In Expt 2, repeated milk samples were taken every 20 s from one quarter during a 120-s teat stimulation in 20 cows with different somatic cell counts (SCC). Quarters were clustered for low (<5·0 log cells/ml), mid (5·0–5·7 log cells/ml) and high (>5·7 log cells/ml) SCC of the sample taken at t=0 s. Samples were analysed for SCC, electrical conductivity (EC) and Na+ and Cl− concentrations. During the experimental inflammation SCC, EC, Na+ and Cl− peaked at 12 h from LPS administration and values in treated quarters (T) at this time were elevated to 7900, 157, 501 and 169% of the values in untreated quarters, respectively. In Expt 2, SCC, EC, Na+ and Cl− in high SCC quarters were 2520, 121, 283 and 141% of low SCC quarters at the start of stimulation (t=0 s), respectively. Highly significant (P<0·001) differences in EC, Na+ and Cl− between high and low SCC quarters disappeared owing to the onset of alveolar milk ejection 100 s after the first contact with the teat. In conclusion, SCC in cows' milk provided the strongest amplitude in the case of an intramammary inflammation. EC, Na+ or Cl− were useful tools only if the measurements were performed in cisternal milk before the start of alveolar milk ejection.

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Assessment of udder health in lame cows

Acta Veterinaria Brno ◽

10.2754/avb202089020133 ◽

2020 ◽

Vol 89 (2) ◽

pp. 133-140

Author(s):

Vladimír Hisira ◽

Pavel Slovák ◽

Paulína Marčeková ◽

Marián Kadaši ◽

Pavol Mudroň

Keyword(s):

Dairy Cows ◽

Subclinical Mastitis ◽

Clinical Mastitis ◽

Coagulase Negative Staphylococci ◽

Digital Dermatitis ◽

White Line ◽

Udder Health ◽

Milk Samples ◽

California Mastitis Test ◽

Sole Ulcers

The objective of this study was to assess the udder health in lame dairy cows. The study was performed on 35 dairy cows which were admitted to the Clinic of Ruminants. The most frequent claw diseases were white line abscess (28.6%) and toe necrosis (28.6%), followed by digital dermatitis (17.1%), toe ulcers (14.3%), and sole ulcers (11.4%). The prevalence of mastitis in lame cows was 74.3% (26); clinical mastitis was detected in 5 (19.2%) and subclinical mastitis in 21 (80.8%) dairy cows. Distribution of mastitis was similar in terms of front (52.54%) and rear quarters (47.46%). For statistical analyses of mastitis grade in lame dairy cows, a six-point Mastitis Score was created based on physical examination of the mammary gland and results of California Mastitis Test. The Mastitis Score tended to be higher in cows with corium inflammation than in those with digital dermatitis (10.0 and 6.5, respectively). Microbiological culture of 24 positive milk samples was performed and intra-mammary pathogens were isolated in 18 (75%) samples. The most prevalent bacteria were coagulase negative staphylococci (50%), followed by Enterococcus spp. (16.7%), Proteus spp. (11.1%), and Aerococcus viridians (11.1%). Of the total bacteria, contagious pathogens were determined in two milk samples (11.1%). In conclusion, high prevalence of mastitis was detected in lame cows suffering from claw diseases without a significant effect of the type of claw diseases on the mastitis grade. Moreover, the dominating isolation of environmental pathogens may be a result of longer lying period of animals affected with lameness.

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A polyol mixture or molasses treated beet pulp in the silage based diet of dairy cows: II. The effect on the lactoperoxidase and thiocyanate content of milk and the udder health

Agricultural and Food Science ◽

10.23986/afsci.71949 ◽

1977 ◽

Vol 49 (5) ◽

pp. 330-345

Author(s):

Hannu Korhonen ◽

Olli Rintamäki ◽

Matti Antila ◽

Mikko Tuori ◽

Esko Poutiainen

Keyword(s):

Dairy Cows ◽

Cell Count ◽

Control Group ◽

Carbohydrate Diet ◽

Cell Content ◽

Mean Values ◽

Udder Health ◽

Milk Samples ◽

Beet Pulp ◽

The Mean

The study investigated the effect of a diet containing either a polyol mixture (polyol group) or molasses (molasses group) on the lactoperoxidase (LP) and thiocyanate (SCN- ) content of milk and the udder health of dairy cows during a 12 week trial period. The control group received no extra carbohydrate feed. On the basis of the weekly milk samples from all test animals the polyol group had on an average the highest LP content (17.8 µg/ml), the lowest SCN- content (0.87 mg/l), and the lowest somatic cell count (152 000 cells/ml). The mean values for the molasses group were; LP: 12.6 µg/ml, SCN- ; 1.01 mg/ml and cell count: 626 000 cells/ml. The same values for the control group were 11,7 µg/ml, 0.91 mg/l and 285 000 cells/ml, respectively. The polyol group yielded milk with an average of 51.5 % more lactoperoxidase daily than the molasses group, and 42.5 % more than the control group. These differences were, however, not found to derive from the different carbohydrate diet, because no significant change in the LP level in any group occurred during the test feeding. The LP and SCN- contents varied considerably from one cow to the other. The degrees of correlation between the factors tested varied markedly among the test groups. The overall values for r were as follows: LP: SCN- = —0.049, LP: cell count = 0.222 and SCN-: cell count = 0.080. On the basis of cell content and the occurrence of mastitis cases, the polyol group had the best and the molasses group the worst udder health. The possible effect of the LP/SCN- /H2O2 antimicrobial system on mastitis resistance is evaluated.

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Relationships among macro-minerals, other selected serum markers of bone profile and milk components of dairy cows during late-lactation

Annals of Animal Science ◽

10.2478/aoas-2020-0094 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Ryszard Mordak ◽

Zbigniew Dobrzański ◽

Robert Kupczyński

Keyword(s):

Dairy Cows ◽

Milk Fat ◽

Dry Matter ◽

Inorganic Phosphorus ◽

Serum Markers ◽

Laboratory Diagnostics ◽

Lactation Period ◽

Milk Samples ◽

Milk Components ◽

Macro Mineral

AbstractTesting blood and milk parameters as well as analysing the relationships among these markers is very useful for monitoring the internal homeostasis and health in high-yielding dairy cows during various production periods. The aim of the study was to assess the correlations (relationships) among macro-minerals, such as calcium (Ca), inorganic phosphorus (P), magnesium (Mg), other selected bone profile markers, such as total protein (TP), albumin, activity of alkaline phosphatase (ALP) measured in serum and selected milk components such as number of somatic cells (SCC), colony-forming units (CFU), milk fat (MF), milk protein (MP), milk lactose (ML), dry matter (DM), non-fat dry matter (FDM) and milk production in late-lactation cows. Both blood and milk samples were collected from 11 clinically healthy milking cows during the late-lactation period. The cows were examined once a day for 3 consecutive days resulting in 33 sets of blood and milk samples for laboratory and statistical analysis. Significant correlations were observed between: Mg and MP, Mg and FDM, ALP and SCC, TP and SCC, TP and MP, TP and FDM, albumin and MP, albumin and FDM, P and Mg, Mg and albumin, and between TP and albumin. When monitoring macro-mineral homeostasis and mammary gland health, especially in intensively fed high-yielding dairy cows correlations between these markers should be considered. The revealed correlations can allow for deeper comparative laboratory diagnostics of homeostasis and can be especially useful for laboratory monitoring of the potential risk of subclinical macro-mineral deficiency in high-yielding dairy cows.

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The Effectiveness of Amoxicillin and Cefalexin in Two Species from Staphylococcus and Bacillus Isolated from Bovine Mastitis in Sudan

Journal of Regenerative Biology and Medicine ◽

10.37191/mapsci-2582-385x-1(1)-004 ◽

2019 ◽

Author(s):

Reem Rabie Mohammed Salih

Keyword(s):

Bacillus Cereus ◽

Bovine Mastitis ◽

Bacillus Coagulans ◽

Staph Aureus ◽

Milk Samples ◽

Sensitivity Tests ◽

Bacillus Spp ◽

Staphylococcus Spp

In this study 60 milk samples collected from mastitic cows to isolates and identify bacterial agents. The percentage of isolates was as follows: Staphylococcus spp 52%, Bacillus spp 26%, Enteroccocispp 3%; microccus 3%, Nesseria 2%, Branhamella 3%, Clostridium 1%, Bordetella 2%, Enterobacteria 3%, Aeromonas 4%, Fusobacterium 1%. In sensitivity tests used two antibiotics Amoxicillin and Cephalexin against two highest isolates in this study (Staph spp and Bacillus spp and found: Staph. aureus and Staph. hyicus were resistant to Amoxicllin and cephlaxin relatively, Bacillus coagulans and Bacillus cereus were susceptible to Amoxicllin and cephlaxin.

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