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2021 ◽  
Author(s):  
Hye-Ri Jung ◽  
Young Ju Lee

Abstract Background: Staphylococcus aureus, a persistent and chronic mastitis-causing pathogen, produces various virulence factors, including enterotoxins. This study analyzed the genetic characteristics of bovine mastitis-related virulence factors to evaluate potential pathogenesis in S. aureus isolated from bulk tank milk.Results: Among 93 S. aureus isolates from 396 dairy farms in six factories operated by three dairy companies in Korea, 40 (43.0%) isolates carried at least one or more enterotoxin genes and there were significant differences between factories within the same company (p < 0.05). Moreover, S. aureus carrying enterotoxin genes showed a higher prevalence in all virulence genes tested in this study except for pvl and lukM, which were not detected in any isolate, than the isolates without enterotoxin genes. In particular, the prevalence of six genes (hla, hlb, lukED, fnbA, clfA, and clfB) was significantly higher in S. aureus carrying enterotoxin genes than isolates without enterotoxin genes (p < 0.05). The most common multilocus sequence type (ST) of 40 enterotoxin-producing isolates was ST188, and all isolates of ST188 harbored the see gene. However, none of the isolates of ST1 and ST72 carried the see gene, and all isolates of ST1 carried the seh gene.Conclusions: Although S. aureus isolated from bulk tank milk, not from mastitis, had a high prevalence of enterotoxins and virulence factors simultaneously, posing a public health threat. Moreover, high enterotoxins in bulk tank milk may be reflected by poor hygiene; therefore, it is important to develop strong monitoring and sanitation programs to ensure that dairy factories produce hygienic milk.


Animals ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3552
Author(s):  
Sabrina de Freitas Vieira ◽  
Sandra Gesteira Coelho ◽  
Hilton do Carmo Diniz Neto ◽  
Hemily Cristina Menezes de Sá ◽  
Barbara Pironi Pereira ◽  
...  

The aim of this study was to evaluate the effects of bulk tank milk (BTM), WM, and PWM on the intake, ruminal parameters, blood parameters, health, and performance of dairy calves. Forty-five male crossbred dairy calves (Gyr × Holstein) were used. On their fourth day of age, animals were grouped according to body weight, serum protein levels, and genetic composition. Three treatments were assessed: BTM (n = 15), WM from cows in antibiotic treatment (n = 15), and PWM via high-temperature, short-time pasteurization (72–74 °C for 16 s) (n = 15). During the experimental period (from 4 to 60 d of age), animals were fed 6 L of milk/d, divided into two equal meals. Water and concentrate were provided ad libitum. Daily measurements were made for milk, concentrate, and water intakes, as well as for fecal and respiratory scores. Rumen fluid and blood were sampled weekly. The following parameters were evaluated: volatile fatty acids (VFAs), pH and ammonia-N in rumen fluid, and β-hydroxybutyrate (BHB) and glucose in blood. Animals were weighed at birth, 4 d of age, and weekly up to 60 d of age. At the end of the experimental period (60 ± 1 d), all animals were euthanized for pulmonary evaluation. The randomized complete design with an interaction between treatment and week was the experimental method of choice for testing the hypothesis of the treatment’s effect on all evaluated outcomes. Animals in the BTM treatment had higher milk dry matter intake (DMI), followed by WM and PWM calves. Concentrate DMI was lower for BTM in comparison to WM and PWM calves. However, total DMI showed no significant differences between treatments. The rumen fluid from calves receiving PWM had higher concentrations of acetate and propionate than that of BTM and WM animals. No differences were observed between treatments for blood glucose and BHB concentrations. Health parameters (fecal and respiratory scores) and pneumonia occurrence showed no significant difference between treatments. No differences were observed for average daily gain (ADG) or body growth. Feeding WM and PWM did not show significant negative effects on the intake, ruminal parameters, blood parameters, health, or performance of dairy calves.


2021 ◽  
Vol 12 ◽  
Author(s):  
Xiaonan Zhao ◽  
Ming Hu ◽  
Cui Zhao ◽  
Qing Zhang ◽  
Lulu Li ◽  
...  

Staphylococcus aureus (S. aureus) is now regarded as a zoonotic agent. Methicillin-susceptible S. aureus (MSSA) ST398 is a livestock-associated bacterium that is most prevalent in China, but there are currently no data available for Shandong. Therefore, the aim of this study was to investigate the epidemiology and characterization of MSSA ST398 from retail pork and bulk tank milk (BTM) in Shandong. A total of 67 S. aureus isolates were collected from retail pork between November 2017 and June 2018. Among the isolates, high antimicrobial resistance rates were observed for penicillin (97.0%), and 92.5% of the isolates were multi-drug resistant (MDR). Eight sequence types (STs) were identified in the retail pork isolates, and the predominant type was ST15 (n=26), which was followed by ST398 (n=14). Staphylococcal protein A gene (spa) typing identified spa types t034 and t1255 in MSSA ST398 from retail pork. Using whole-genome sequencing analysis, we described the phylogeny of 29 MSSA ST398 isolates that were obtained from retail pork (n=14) and BTM (n=15). The phylogenetic tree showed that the MSSA ST398 isolates from different sources had the same lineage. Among the 29 MSSA ST398 isolates, five resistance genes were detected, and all isolates carried DHA-1. Fifteen toxin genes were detected, and all isolates carried eta, hla, and hlb. In conclusion, this study found that a high risk for MSSA ST398 was present in retail pork and BTM. These findings have major implications for how investigations of MSSA ST398 outbreaks should be conducted in the One-Health context.


2021 ◽  
Vol 8 ◽  
Author(s):  
Marie-Claire McCarthy ◽  
Luke O'Grady ◽  
Connor G. McAloon ◽  
John F. Mee

Since the abolition of EU milk production quotas in 2015, Europe's dairy industries have undergone a period of rapid expansion with possible resultant increased inter-herd transmission of endemic pathogens. The aims of this study were (1) to establish the post-2015 prevalence of antibodies to selected endemic infectious diseases and (2) to determine if prevalences differed between herds where heifers were reared at home and those where heifers were sent out for contract-rearing. Three bulk tank milk (BTM) samples were collected annually between May and August of 2018–20 inclusively from 120 Irish dairy herds. Additionally, herd vaccination status was collected by questionnaire. Milk samples were tested using commercially available ELISAs for eight pathogens: bovine viral diarrhea virus (BVDV), bovine herpesvirus 1 (BoHv-1), bovine respiratory syncytial virus (BRSV), Mycoplasma bovis, Mycobacterium avium subspecies paratuberculosis (MAP), Salmonella Dublin (S. Dublin), Leptospira Hardjo (L. Hardjo), and Neospora caninum (N. caninum). The true prevalence of each pathogen was calculated using a Rogan-Gladen estimator. The true prevalences (95% CI) of BTM antibodies in unvaccinated herds across the 3 years were as follows (i) BVDV: 57, 86, and 73% (95% CI: 40.7–65.9, 74–94, and 58–85) (n = 56, 56, and 48), (ii) BoHv-1: 47, 49, and 19% (95% CI: 26.3–69.7, 25–75, and 1–56) (n = 21, 20, and 11), (iii) L. Hardjo: 34, 59, and 73% (95% CI: 12.5–63, 33–82, and 33–99) (n = 15, 21, and 10), (iv) S. Dublin 32, 57, and 11% (95% CI: 12.21–68.1, 30.2–90.1, and 0) (n = 19, 22, and 13), (v) BRSV: 100% (95% CI: 99.5–100, 100, and 100) (n = 120, 109, and 91), (vi) MAP: 0% (95% CI: 0, 0, and 0) (n = 120, 109, and 91) (vii) N. caninum 0% (95% CI: 0, 0, and 0) (n = 120, 109, and 91) and (viii) M. bovis (ELISA) 53, 0.42, and 30% (95% CI: 3.95–6.84, 0, and 21–41) (n = 120, 109, and 91). M. bovis was detected by PCR in 0, 1, and 0% of herds in 2018, 2019, and 2020, respectively. This study showed that expanding Irish dairy herds are endemically infected with several of the studied pathogens. No differences in herd prevalence of infectious agents were observed between farms with different heifer rearing strategies (contract-rearing vs. traditional rearing).


2021 ◽  
Author(s):  
Lene Idland ◽  
Erik G. Granquist ◽  
Marina Aspholm ◽  
Toril Lindbäck

Abstract Aims: This study explores how different dairy farm operating systems influence the occurrence of zoonotic bacteria in raw milk. Methods and Results: Samples from bulk tank milk, milk filters, feces, feed, teats and teat milk were collected from eleven farms with loose housing and seven with tie-stall housing every second month over a period of 11 months and analyzed for the presence of Campylobacter spp., L. monocytogenes and STEC. Campylobacter spp., L. monocytogenes and STEC were abundant in samples from the farm environment and were also detected in 4%, 13% and 7% of the milk filters, respectively, and in 3%, 0% and 1% of bulk tank milk samples. Four STEC isolates carried the eae gene, which is linked to the capacity to cause more severe human disease. Conclusion: The results indicate a higher prevalence of L. monocytogenes and Campylobacter spp. in samples collected from loose housed herds compared to tie-stalled herds suggesting that the operating system can influence the food safety of raw milk. Significance and Impact of the study: This study highlights that zoonotic bacteria can be present in raw milk independent of hygienic conditions at the farm and what hosing system is used. Altogether, this study provides an important knowledge base for evaluating the risk of drinking unpasteurized milk.


Biology ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 1016
Author(s):  
Daphne T. Lianou ◽  
Efthymia Petinaki ◽  
Peter J. Cripps ◽  
Dimitris A. Gougoulis ◽  
Charalambia K. Michael ◽  
...  

The objectives of this work were to study prevalence and characteristics of resistance to antibiotics of staphylococcal isolates from the bulk-tank milk of sheep flocks across Greece, to assess possible associations of the presence of antibiotic resistance with the quality of milk in these flocks and to evaluate flock-related factors potentially associated with antibiotic resistance among these isolates. A cross-sectional study was performed in 325 sheep flocks in Greece. Bulk-tank milk samples were collected for bacteriological examination; staphylococcal isolates were evaluated for resistance to 20 antibiotics. Oxacillin-resistant staphylococcal isolates, isolates resistant to any antibiotic, and multi-resistant isolates were recovered from 8.0%, 30.5%, and 12.0% of flocks, respectively. Of 232 isolates, 11.6% were resistant to oxacillin, 46.1% were resistant to at least one antibiotic, and 16.4% were multi-resistant. Resistance was seen more frequently among coagulase-negative (50.6%) than among Staphylococcus aureus (31.5%) isolates. Resistance was more frequent against penicillin and ampicillin (34.1% of isolates), clindamycin (17.7%), and fosfomycin (14.2%). An association was found between biofilm formation by staphylococci and resistance to fosfomycin. For recovery of oxacillin-resistant isolates, the lack of experience by farmers emerged as a significant factor; respective factors for the isolation of staphylococci resistant to any antibiotic or multi-resistant isolates were the early stage of the lactation period (0th–1st month) and the intensive management system applied in the flocks, respectively.


Antibiotics ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 1225
Author(s):  
Daphne T. Lianou ◽  
Efthymia Petinaki ◽  
Peter J. Cripps ◽  
Dimitris A. Gougoulis ◽  
Charalambia K. Michael ◽  
...  

The objectives of this work were to study the prevalence and the patterns of antibiotic resistance of staphylococcal isolates from bulk-tank milk of goat herds across Greece, to assess possible associations of the presence of antibiotic resistance with the quality of milk in these herds and to evaluate herd-related factors potentially associated with the presence of antibiotic resistance among these staphylococcal isolates. A cross-sectional study was performed on 119 goat herds in Greece. Bulk-tank milk samples were collected for bacteriological examination; staphylococcal isolates were evaluated for resistance to 20 antibiotics. Oxacillin-resistant, resistant to at least one antibiotic, and multi-resistant staphylococcal isolates were recovered from 5.0%, 30.3%, and 16.0% of herds, respectively. Of 80 isolates, 7.5% were resistant to oxacillin, 50.0% were resistant to at least one antibiotic and 27.5% were multi-resistant. Resistance was seen more frequently among coagulase-negative staphylococci (59.3%) than among Staphylococcus aureus (23.8%). Resistance was more frequent against penicillin and ampicillin (41.3% of isolates) and fosfomycin (27.5%). No association was found with biofilm formation by staphylococci. For recovery of oxacillin-resistant isolates, the presence of working staff in the herds emerged as a significant factor; respective factors for the isolation of staphylococci resistant to at least one antibiotic were part-time farming and high (>10) number of systemic disinfections in the farm annually. The same three factors concurrently were also identified to be significant for the recovery of multi-resistant isolates.


2021 ◽  
Vol 74 (1) ◽  
Author(s):  
Hye-Ri Jung ◽  
Koeun Kim ◽  
Young Ju Lee

Abstract Background This study was conducted to analyze the genetic characteristics of 41 β-lactam-resistant Escherichia coli isolates, which are one of the common causes of environmental mastitis, isolated from the bulk tank milk of 290 dairy farms in five factories operated by three dairy companies in Korea. Results Analysis of the phenotypic and genotypic characteristics of β-lactam-resistant E. coli isolates revealed differences between factories even within the same company. Isolates from factory A1 and C1 showed high resistance to cephalothin (76.9 and 100%, respectively), which is a first-generation cephalosporins, whereas resistance to tetracycline was showed by only the isolates from factories B1 (60.0%), C2 (66.7%), and C3 (100%). Although all the 41 β-lactam-resistant E. coli isolates were positive for blaOXA-1, blaTEM-1 was highly prevalent in isolates from factories C2 (100%) and C3 (100%). Among 17 isolates resistant to both β-lactams and aminoglycosides, the most common multilocus sequence type was ST399 (13isolates, 76.5%). Furthermore, 2 (11.8%) and 12 (70.6%) isolates belonged to the phylogenetic groups B2 and D, respectively, which are invasive strains that cause intestinal infections, respectively. The predominant serogroup was O15 (70.6%), which is a globally distributed extraintestinal pathogen. Interestingly, one isolate from factory A1 belonged to O157 and carried six virulence genes, simultaneously. Conclusions Although E. coli isolates were isolated from bulk tank milk, and not the clinical mastitis samples, the presence of the phylogenetic groups B2 and D, and the serogroups O15 and O157, which harbor antimicrobial resistance genes and virulence factors, can pose a threat to public health.


Author(s):  
Min Yap ◽  
David Gleeson ◽  
Paul W O’Toole ◽  
Orla O’Sullivan ◽  
Paul D. Cotter

Cleaning of the production environment is vital to ensure the safety and quality of dairy products. Although cleaning with chlorine-based agents is widely adopted, it has been associated with detrimental effects on milk quality and safety, which has garnered increasing interest in chlorine-free cleaning. However, the influence of these methods on the milk microbiota is not well documented. This study investigated the factors that influence the raw milk microbiota, with a focus on the differences when chlorine-based and chlorine-free cleaning of milking equipment are used. Bulk tank raw milk was sampled at three sampling months (Apr, Aug and Nov), from farms across Ireland selected to capture the use of different cleaning methods, i.e., exclusively chlorine-based (n = 51) and chlorine-free cleaning (n = 92), and farms that used chlorine-free agents for the bulk tank and chlorine-based cleaning agents for the rest of the equipment (n = 28). Shotgun metagenomic analysis revealed the significant influence of seasonal and geographic factors on the bulk tank milk microbiota, indicated by differences in diversity, taxonomic composition, and functional characteristics. Taxonomic and functional profiles of samples collected in November clustered separately from other months. In contrast, cleaning methods only accounted for 1% of the variation in the bulk tank milk bacterial community, and samples collected from farms using chlorine relative to chlorine-free cleaning did not differ significantly, suggesting that chlorine-free approaches used did not negatively impact microbiological quality. This study shows the value of shotgun metagenomics in advancing our knowledge of the raw milk microbiota. IMPORTANCE The microbiota of raw milk is affected by many factors that can control or promote the introduction of undesirable microorganisms. Chlorine-based cleaning agents have been commonly used due to their effectiveness in controlling undesirable microorganisms, but have been associated with the formation of chlorine residues that are detrimental to product quality and may impact consumer health. Chlorine-free alternatives have been recommended in some countries, but the influence of cleaning agents on the milk microbiota is unknown. Here we investigated the influence of cleaning methods and other factors on bulk tank raw milk. Results showed that season and location had a greater influence on the milk microbiota than the cleaning agents used. Indeed, the similar microbiota compositions of raw milk from farms that used chlorine-based and those that used chlorine-free cleaning methods supports the further use of chlorine-free cleaning agents in dairy production.


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