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Council Directive 92/46/EEC. Laying down the health rules for the production and placing on the market of raw milk, heat- treated milk and milk-based products. 2004. Bava L, Zucali M, Sandrucci A, Brasca M, Vanoni L, Zanini L, Tamburini A. Effect of cleaning procedure and hygienic condition of milking equipment on bacterial count of bulk tank milk. J Dairy Res. 2011;78:211–9. https://doi.org/10.1017/S002202991100001X. This study aimed to investigate the temporal dynamics of the microbiota in the bovine udder. The ten cows used for this purpose were divided in two groups based on the level of SCC before the first sampling. Five of the cows (L1–L5) had a stable low SCC (< 100,000 SCC/mL) on the three days leading up to the first sampling, while the remaining five cows (H1–H5) had a higher SCC (> 100,000 SCC/mL) in the same period. Quarter milk samples were collected from all ten cows at six samplings during a period of five months (January to May). Of the 240 quarter samples that were collected, six were missing during collection and were not included in the analysis. To study the microbial composition of the 234 remaining samples, amplicon sequencing of the 16S rRNA genes were performed for all the samples. The average depth of sequencing was 49,093 sequences per sample before filtering and 18,880 sequences per sample after filtering. In total 9132 high quality SVs were obtained from 234 samples. 14 samples were filtered out of the analysis because they did not pass the quality filtering of the Dada2 pipeline used to analyse the 16S data after sequencing. Of the 9,132 high quality SVs, 6962 SVs were used for taxonomy search, and 553 SVs were successfully assigned to family level. 61 quarter samples were classified as having an IMI based on definition “A” from Dohoo et al. [ 23]. This definition states that a quarter sample where > 10 colonies are cultured per 0.1 mL is defined as having an IMI. Eighty-nine percent of these were from group H and 11% were from group L. A limit of 100,000 SCC/mL was selected to classify the quarter samples as high or low SCC during samplings. 30 quarter samples had high SCC, 204 low SCC, and 6 samples had no recorded SCC. Of the 30 samples with a recorded high SCC, 93% were from group H and 7% from group L. Other additional data about the health, parturition, and recorded mastitis were retrieved from the Norwegian Cattle Health Recording System (Additional file 1). No cows were recorded to have mastitis during the sampling period, while two cows (L1 and H2) were treated for mastitis caused by Streptococcus dysgalactiae and Streptococcus uberis, respectively, after this period. The samplings occurred between 19 and 193 days in milk for the 10 cows (Additional file 2). This period encompasses the early and mid-lactation stages. Five of the cows (H4, H5, L3, L4, L5) were in their first parturition, while the remaining five (H1, H2, H3, L1, L2) were in their second parturition. Diversity analysis of the quarter samples
Tarekgne E, Skeie S, Rudi K, Skjerdal T, Narvhus JA. Staphylococcus aureus and other Staphylococcus species in milk and milk products from Tigray region, Northern Ethiopia. Afr J Food Sci. 2015;9:567–76. Mastitis is characterized by mild or severe pathological changes in udder tissue, increased somatic cells count (SCC) and abnormal milk quality ( 4). Dairy cow mastitis is the most expensive infectious disease in dairy cow breeding worldwide. In terms of the impact on animal husbandry in the United States alone, the annual losses caused by the decline of milk production and quality, the soaring cost of veterinary treatment and the increasing cost of farm management are as high as billions of dollars ( 5). Cow mastitis is usually divided into clinical mastitis and subclinical mastitis according to the visible changes of udder and milk ( 6), and there are 137 kinds of pathogenic bacteria in dairy cow mastitis ( 7, 8), and Staphylococcus aureus, Escherichia coli, Streptococcus are the most common pathogens that cause mastitis ( 9, 10). The correlation between TBC and CC (r = 0.5) in this study corroborated previous studies that reported a moderate to high correlation [ 13, 15, 56, 69]. This indicates that TBC could be used as a single microbiological hygienic indicator for total aerobic bacteria present in raw milk [ 15]. Mild mastitis: Abnormality of the milk is the main sign with little evidence of change in the udder and no systemic signs such as dullness and loss of appetite.Hohmann M, Wente N, Zhang Y, Krömker V. Bacterial load of the teat apex skin and associated factors at herd level. Animals. 2020;10:1647. For the analysis of the microbiota, the bacterial pellet was obtained from 40 mL of each quarter milk sample, as previously described [ 2]. Briefly, 40 mL of milk was centrifuged at 8000× g for 10 min, the fat layer removed with a sterile cotton swab, and the supernatant removed. The pellet was then washed twice with 2% citrate water, and DNA was extracted from each pellet using the DNeasy PowerFood Microbial Kit (Qiagen, Düsseldorf, Germany) starting from step 3 in the detailed protocol of DNeasy Powerfood Microbial Kit Handbook. For increased efficiency of lysis of difficult species additional 5 min of vortex time were added to step 6, bringing the total vortex time to 15 min. DNA was finally eluted in 50 µl elution buffer before storage at − 20 °C. Library preparation for amplicon sequencing using the Illumina Miseq platform was performed as described previously [ 2]. The V3 and V4 region of the 16S rRNA was amplified using the primers Uni340F (CCTACGGGRBGCASCAG) and Bac806R (GGACTACYVGGGTATCTAAT). PCR reagents and conditions were identical to the one described by Porcellato et al. [ 2]. Negative controls were included to monitor for contamination during DNA extraction and during library preparation. The final library concentration was then measured using Qubit 2 with the dsDNA HS kit (ThermoFischer Scientific) and quantitated using the KAPA Library Quantification kit (Illumina) before being sequenced on an Illumina MiSeq platform (Illumina) using the 2 × 300 bp V3 kit (Illumina). Sequence data analysis and statistical testing
Millogo V, Sjaunja KS, Ouédraogo GA, Agenäs S. Raw milk hygiene at farms, processing units and local markets in Burkina Faso. Food Control. 2010;21:1070–4. Our Skin Milk Udder Cream is also used in Oncology hospitals and clinics across Australia as a moisturising cream for dry and damaged skin following radiation treatment and chemotherapy.Fig 5: There is little sense in treating an older cow with chronic Staph aureus and a persistently high cell count Antibiotics All farms should have an individualised standard operating procedure (SOP) for the treatment of mastitis with antibiotics. This should include details such as when to use intramammary antibiotics, when to use systemic therapy and when to use combined treatment, as well as guidance on when not to treat because the cow is unlikely to respond (a high SCC cow with a history of Staph. aureus infection). It should also detail the products available to the staff that have been prescribed for mastitis treatment on that farm. All staff milking cows should be familiar with this SOP. Hamiroune M, Berber A, Boubekeur S, Smar O. Evaluation of the bacteriological quality of raw cow’s milk at various stages of the milk production chain on farms in Algeria. Rev Sci Tech Off Int Epiz. 2016;35:1–26. Piepers S, Zrimšek P, Passchyn P, Vliegher S De. Manageable risk factors associated with bacterial and coliform counts in unpasteurized bulk milk in Flemish dairy herds. J Dairy Sci. 2014;97:3409–19. http://dx.doi.org/10.3168/jds.2013-7203. The hygiene of the cows was evaluated based on the visual cleanliness scores described by Schreiner and Ruegg [ 39]. Evaluation was performed in the legs, flanks, abdomen, and udder of each animal. Score 1 (VC) indicates very clean, score 2 (C) indicates clean, score 3 (D) indicates dirty, and score 4 (VD) indicates very dirty. Farms were considered clean if the number of milking cows with cleanliness scores of VC and C were equal to or more than 50% of the total milking cows in the herd and taken as dirty if the number of milking cows with cleanliness scores of D and VD were equal to or more than 50% of the total milking cows in the herd. Udder scoring followed the scoring system described by Schreiner and Ruegg [ 39], where score 1 was free of dirt, score 2 was slightly dirty (2–10% of surface area), score 3 was moderately covered with dirt (10–30% surface area) and score 4 was covered with caked on dirt (> 30% of surface area). Enumeration of contaminating bacteria
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