Bulk Tank Milk Research Articles

Comparisons of recycled manure solids and wood shavings/sawdust as bedding material—Implications for animal welfare, herd health, milk quality, and bedding costs in Swedish dairy herds

Increasing shortages and costs of common bedding materials have led dairy farmers in Sweden to consider using recycled manure solids (RMS), which are readily available and low cost, as an alternative bedding material. The main risks are effects on udder health and milk quality, but RMS could also affect animal welfare and claw health. The advantages and disadvantages of using RMS bedding have not been fully investigated, and findings in other countries cannot be directly applied to Swedish conditions and climate. This observational cross-sectional study investigated the use of RMS as bedding, regarding associations with certain aspects of animal welfare, herd health, milk quality, and bedding costs in Swedish dairy herds. Thirty-four dairy farms using RMS or wood shavings/sawdust (each n = 17) were compared. Each farm was visited 2 times during the housing period from 2020 to 2021, once from October to December and once from March to May. Dairy barns were observed, animal welfare was assessed, and freestall dimensions were measured. Farm owners were interviewed about housing system characteristics, herd performance, and herd management. Data on milk production and herd health were obtained from the Swedish official milk recording scheme for the indoor period from October to March. The prevalence of claw disorders and abnormal claw conformation were collected from the national claw health database for the period from October to May. On each farm visit, composite samples of unused bedding outside the barn and used bedding material from the freestalls, respectively, were taken for total bacterial count and DM analysis. Samples of bulk tank milk for determination of total bacterial count were taken in connection to the visits. In addition, samples of unused and used bedding material and manure from alleys for analysis of 3 Treponema species associated with digital dermatitis (DD) were gathered and analyzed. Total bacterial count was significantly higher in unused (8.50 log10 cfu/g) and used RMS bedding (9.75 log10 cfu/g) than in wood shavings/sawdust (used 4.74; unused 8.63 log10 cfu/g), but there were no significant differences in bulk milk total bacterial count (median 4.07 vs. 3.89 log10 cfu/mL) or SCC (median 243,800 vs. 229,200 cells/mL). The aspects of animal welfare assessed did not differ significantly between the 2 bedding systems, whereas the prevalence of total claw disorders (25.9% vs. 38.0% of trimmed cows), dermatitis (6.9% vs. 16.2% of trimmed cows) and sole ulcers (2.0% vs. 4.0% of trimmed cows) were significantly lower in the RMS herds. Treponema spp. were not detected in unused RMS material, but all RMS herds had presence of DD recorded at foot trimming. An economic assessment based on the interview results and price level from winter 2021 revealed that the costs of RMS bedding varied with amount of RMS produced. Thus, RMS is a potential alternative bedding material for dairy cows in Sweden and can be a profitable option for large dairy herds. However, the high level of total bacteria in the material requires attention to bedding and milking routines as well as regular monitoring of herd health.

The impact of different temperatures on spore count and browning of UHT milk over a long storage period

One of the major microbiological challenges for the ultra-high temperature (UHT) milk industry is the presence of sporeformers in milk, which can survive pasteurization and the UHT process. The present study has aimed to investigate the spore populations in UHT milk stored at 20 °C and 35 °C for 14 months. Samples of bulk tank milk, pasteurized milk, and UHT milk were randomly assigned to different temperatures and storage times and were analyzed to study the mesophilic populations of aerobic spores and any color changes. The results showed that aerobic sporeformers can survive the UHT process. Bacillus licheniformis group and Bacillus pumilus group were the most isolated aerobic sporeformers. The b∗ value increased significantly in samples stored at 35 °C. These data provide information on the aerobic spore populations of bulk tank milk and UHT milk produced in different seasons and stored at different temperatures for 14 months.

Prevalence of Contagious Mastitis Pathogens in Bulk Tank Milk from Dairy Farms in Lower Saxony, Germany

The aim of this study was to determine the prevalence of Streptococcus (Sc.) agalactiae, Prototheca spp., Staphylococcus (S.) aureus, and especially methicillin-resistant S. aureus as well as Myco-plasmopsis (M.) spp. and M. bovis in bulk tank milk (BTM) on dairy farms in Lower Saxony, Germany. BTM samples were collected in January 2023 from 208 selected dairy farms. The samples were quantitatively culturally analyzed for S. aureus and Prototheca spp. Presumptive S. aureus colonies were further confirmed by MALDI-TOF. Presumptive Prototheca spp. colonies were confirmed by light microscopy. Sc. agalactiae and Mycoplasmopsis spp. were detected by real-time polymerase chain reaction (rtPCR). Sc. agalactiae was detected in two herds (1% (Confidence Interval 95% (CI) 0.3–3.4)). S. aureus was confirmed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) in 38 herds (18.3% (CI 13.6–24.1)), assuming a threshold of >10 cfu/mL milk. A total of 154 isolates identified as S. aureus by MALDI-TOF were transferred to agar with added oxacillin for resistance testing, of which 19 isolates (12.3% (CI 8–18.5)) showed growth. The 19 isolates came from eight different farms (3.8% (2–7.4)). Prototheca spp. were identified in 13 herds (6.3% (CI 3.7–10.4)). Mycoplasmopsis spp. were detected by PCR in 18 herds (8.7% (CI 5.5–13.3)). Of these, M. bovis was present in three herds (1.4% (0.5–4.2)). The herd prevalence of Sc. agalactiae in BTM appears to be at low levels in the sampled area. The prevalence of Mycoplasmopsis spp. in the herds was higher than expected compared to previous studies. It is interesting to note that the percentage of M. bovis in the total Mycoplasmopsis spp. was only 16.7%.

Isolation and antimicrobial resistance patterns of Methicillin-resistantStaphylococcus aureus from raw cow's milk in dairy farms of Wolaita Sodo Town, Southwest, Ethiopia.

Staphylococcus aureus is a pathogenic bacterium-contaminating milk and milk products causing food poisoning primarily due to its enterotoxins. A cross-sectional study was conducted from November 2021 to June 2022 in Wolaita Sodo Town, to detect Methicillin-resistant Staphylococcus aureus (MRSA) in raw cow's milk and assess their resistance levels to different antimicrobials. Purposive sampling was used to select 34 dairy farms. Accordingly, 419 raw milk samples from the farm and collection centers were collected. Isolates of S. aureus showing resistance to Cefoxitin were classified as MRSA. From the total 419 samples, 22.19% (93/419) were contaminated with S. aureus in dairy cows. The prevalence of S. aureus in raw milk, bulk milk from the farm, and bulk tank milk from the collection centers was 16.9%, 2.1%, and 3.1%, respectively. The risk of S. aureus contamination in dairy farm owners and milkers who do not take food safety training was 5.303 times higher than the risk of S. aureus contamination in dairy farm owners and milkers who take food safety training. The risk of S. aureus contamination in dairy farms kept under poor management system was 7.34 times more than that of dairy farms kept under good management. The cefoxitin disk diffusion method was used to detect MRSA, 57.14% being resistant to Cefoxitin in total while approximately 87.5% were sensitive to Sulfamethoxazole-trimethoprim and Gentamycin, while Erythromycin registered 75%, Ciprofloxacin 62.5%, Chloramphenicol 62.5%, and Tetracycline 25%. This study revealed that MRSA exhibited a notable multidrug resistance pattern, encompassing resistance to multiple drugs, with a prevalence of 75%. Significantly, the hands of milkers and the milking containers emerged as pivotal sources of contamination. This underscores the crucial importance of maintaining stringent hygienic practices during the milking process, with particular emphasis on thorough cleaning and decontamination of utensils.

Association of clinical respiratory disease signs and lower respiratory tract bacterial pathogens with systemic inflammatory response in preweaning dairy calves

This study investigated the potential associations between 3 acute phase proteins (APP)-haptoglobin, serum amyloid A-and fibrinogen, clinical signs of respiratory disease, and the presence of bacterial pathogens in the lower respiratory tract (LRT) of preweaning dairy calves. This cross-sectional study included 150 preweaning calves (2-86 d old) from 15 large dairy herds in Estonia. Tracheobronchial lavage, blood, and fecal samples were collected from 5 calves showing clinical signs indicative of LRT disease, and samples from 5 calves without clinical signs of LRT disease per herd. All samples collected from these calves were analyzed for concentrations of systemic APP, LRT bacteria, and intestinal pathogens. Heifer blood and bulk tank milk samples were collected for the detection of disease-specific antibodies against bovine herpesvirus 1, bovine viral diarrhea virus, bovine respiratory syncytial virus, and Mycoplasma bovis. Mixed-effects linear regression models were used to analyze the associations of clinical respiratory disease signs and LRT bacteria with APP. Increased plasma fibrinogen concentrations in calves were associated with higher rectal temperature (>39.5°C), increased respiratory rate (>50 breaths/min), and coughing. Increased serum amyloid A concentrations were associated with higher rectal temperature (>39.5°C) and respiratory rate between 40 and 50 breaths/min. Calves with the presence of fecal Cryptosporidium spp. and rectal temperature of 39°C and above had increased serum haptoglobin concentrations. Increased fibrinogen concentrations were associated with the presence of Pasteurella multocida in the calf LRT, whereas increased concentrations of fibrinogen and serum amyloid A were associated with the presence of Trueperella pyogenes. In conclusion, APP showed variable associations with clinical signs of respiratory disease and LRT bacteria. Plasma fibrinogen concentration could be used as a complementary calf-side test to assess systemic inflammation caused by LRT bacteria such as Pasteurella multocida and Trueperella pyogenes in preweaning dairy calves.

Neospora caninum antibodies in bulk tank milk from dairy cattle herds in Italy in relation to reproductive and productive parameters and spatial analysis

Among the available diagnostic techniques, antibody detection in bulk tank milk (BTM) represents a useful tool to estimate and monitor Neospora caninum herd prevalence.To evaluate the prevalence of N. caninum and the effect of parasite infection on herd performances, BTM samples collected from 586 dairy herds located in one of the largest dairy production areas in Italy (Lombardy) were analyzed by an indirect ELISA to detect anti-N. caninum specific antibodies. Generalized linear models (GLMs) were developed. A purely spatial analysis scanning for clusters with high or low rates for N. caninum using the Bernoulli model was performed. A maximum entropy approach was used to estimate the probability of distribution of the parasite based on occurrence records together with environmental variables.Overall, 180 herds resulted positive for N. caninum antibodies on bulk tank milk (P = 30.7 %). A higher risk of seropositivity was evidenced in the provinces of Milano, Cremona, Brescia, and Bergamo (P = 32–40 %); a lower risk was evidenced in Lodi, Pavia, and Mantova (P = 13–24 %). A higher risk of seropositivity was revealed for small-medium farms (101–300 animals) (O.R.=2.8) and for older animals with more than 4 years (O.R.=4.4).Regarding the effect of N. caninum infection on herd performances, the number of inseminations for conception was higher (> 3 inseminations), and the period from calving to conception was longer (> 150 days) for positive farms (O.R.=2.0 and O.R.=2.3, respectively); besides, lower head daily milk production (<20 kg and 21–25 kg) and mature equivalent milk yield (<11,000), and somatic cell counts higher than 300,000 cells/ml were observed for N. caninum positive herds (O.R.=0.4, O.R.=0.4 and O.R.=1.9 respectively).The geographical distribution of N. caninum positive farms with the highest level of probability covers the central sector of the Po Plain where a significant cluster for high risk of parasite infection was shown by spatial scan statistic and Maximum entropy ecological niche modelling. A further significant cluster of low risk occurred in the southern. The climatic and environmental variables with the highest training gain when used in isolation resulted altitude, land use/land cover, and other variables related to temperature and precipitation.Neosporosis is widely distributed in Italian dairy herds and an impact of the parasite on herd performances could be hypothesized. Even if the role of N. caninum in alterations of reproductive and productive parameters should be further explored, veterinarians and farmers should be aware of neosporosis, and control plans should be adopted.

Prevalence and associated risk factors of anti-Coxiella burnetii antibodies in dairy cattle herds using bulk tank milk analysis in Kabylia area, north Algeria.

Coxiella burnetii, or Q fever agent, has notable implications for human and livestock health. Infections in cattle primarily manifest through reproductive issues where infected animals shed the bacterium in birth fluids, placental tissues, and milk, serving as potential sources of transmission. Bovine herds become reservoirs, contributing to the environmental contamination of farming areas. Comprehensive studies on the prevalence, transmission routes, and associated risk factors among cattle contribute to the development of effective control strategies, ultimately safeguarding both livestock and public health.Here we determine the prevalence of Coxiella burnetii antibodies against in dairy cattle farms from Kabylia (northern Algeria) and identify the associated risk factors. Bulk tank milk samples from 184 farms were analyzed by indirect ELISA technique, 49 of them were tested positive which corresponds to a prevalence rate of 26.63% (95% CI 20.25-33.01%). Multivariate analysis by logistic regression showed that the risk factors associated with detection of anti-Coxiella burnetii antibodies are: cohabitation of cattle with small ruminants(OR = 3.74 95% CI [1.41-8.92]), exposure to prevailing winds (OR = 5.12 95% CI [2.11-13.45]), and the veterinarian visits frequency(OR = 5.67 95% CI [2.55-13.60]). These findings underscore the susceptibility of dairy cattle to Q fever in the Kabylia region, highlighting practices that pose risks. We recommend the implementation of hygienic measures and adherence to proper farming conditions to mitigate the transmission of Q fever and reduce the associated zoonotic risk.

Qualitative characteristics of sheep's and goat's milk in Albania.

Cost-effective methods are crucial in small-scale operations to assess the physicochemical properties of milk, identify potential clinical or subclinical mastitis, and test β-lactams in small ruminants. The aim was to evaluate the importance of physicochemical characteristics, somatic cells, and antibacterial substances as factors for monitoring the quality and safety of bulk tank milk from sheep and goats. Furthermore, we aimed to elucidate the influence of grazing capacity on the composition of sheep milk. The fat content of sheep milk (n=119) was found to be 7.7%, with a range of 5.3% to 9.9%. The average fat content of dairy goats (n=33) was found to be 4.9%, with a range of 2.8-6.7%. The results indicate that 2.8% of sheep milk samples and 7.8% of goat milk samples exhibited clear positive reactions when assessed using the California mastitis test. β-lactam antibiotics were present in 0.7% of sheep milk samples, but not in any of the goat milk samples using lateral flow strips. This measurement method can be useful to detect adulterated milk and better understand the quality and safety of small ruminant milk before processing.

Associations between Milk Coagulation Properties and Microbiological Quality in Sheep Bulk Tank Milk.

This study conducted a seasonal analysis of bulk tank milk from 77 sheep farms to establish relationships between the concentration of major microbial groups and milk coagulation properties. The investigated milk traits included composition (pH, fat, casein, lactose), coagulation properties (curd firmness: A60-, rennet clotting time: RCT-, curd firming time: k20-, curd yield: CY-), and somatic cell score (SCS). The main microbial groups analyzed were total mesophilic bacteria (SPC), thermodurics (THERMO), psychrotrophs (PSYCHRO), Pseudomonas spp. (PSEUDO), lactic acid bacteria (LAB), catalase-negative gram-positive cocci (GPCNC), Escherichia coli (ECOLI), coliforms other than Escherichia coli (COLI), coagulase-positive staphylococci (CPS), coagulase-negative staphylococci (CNS), and spores of lactate-fermenting Clostridium (BAB). Mixed linear models were used to explore associations between coagulation properties and the aforementioned variables. Results demonstrated that incorporating microbial loads into the models improves their fit and the relative quality of the outcomes. An important seasonality is demonstrated by an increase in CY and A60, along with a decrease in RCT and k20 during autumn and winter, contrasting with spring and summer. BAB concentration resulted in a reduction of A60 and an increase in RCT, whereas SPC concentration led to an enhancement of A60 and a reduction in RCT. An increase in GPCNC concentration was associated with an increase in k20 and a decrease in CY.

Diagnostic performance of Mycoplasmopsis bovis antibody ELISA tests on bulk tank milk from dairy herds

BackgroundTesting of bulk tank milk (BTM) for Mycoplasmopsis bovis (previously Mycoplasma bovis) antibodies is increasingly popular. However the performance of some commercially available tests is unknown, and cutoff values possibly need to be adjusted in light of the purpose. Therefore, the aim of this study was to compare the diagnostic performance of three commercially available M. bovis antibody ELISAs on BTM, and to explore optimal cutoff values for screening purposes. A prospective diagnostic test accuracy study was performed on 156 BTM samples from Belgian and Swiss dairy farms using Bayesian Latent Class Analysis. Samples were initially classified using manufacturer cutoff values, followed by generated values.ResultsFollowing the manufacturer’s guidelines, sensitivity of 91.4%, 25.6%, 69.2%, and specificity of 67.2%, 96.8%, 85.8% were observed for ID-screen, Bio K432, and Bio K302, respectively. Optimization of cutoffs resulted in a sensitivity of 89.0%, 82.0%, and 85.5%, and a specificity of 83.4%, 75.1%, 77.2%, respectively.ConclusionsThe ID-screen showed the highest diagnostic performance after optimization of cutoff values, and could be useful for screening. Both Bio-X tests may be of value for diagnostic or confirmation purposes due to their high specificity.

Energy-Related Hormones in Raw and Retail Cow’s Milk and Possible Risk for Consumers

Abstract The study aimed to determine the concentration of energy-related hormones in cow’s milk and to consider them from a public health perspective. Fourteen Holstein cows were subjected to milk and blood sampling on the following days in lactation (DIL): 10, 30, 60, 90, 150, 180, 210, 250 and 280 to determine milk hormones, fat and protein content and blood biochemical parameters. For the same purpose, bulk-tank milk was sampled and samples of retail m ilk with 1.5% (CM1.5) and 3.2% (CM3.2) fat was purchased. Milk insulin-like growth factor-1 (IGF-1) values were significantly lower at 90, 150, 180, 210 and 250 and significantly higher at 10, 30 and 60 DIL than lactation average (LA). Milk insulin concentrations were significantly lower at 30, 60 and 90 and higher at 210, 250 and 280 DIL than LA. Free thyroxine (fT4) level in the milk was higher at 250 DIL, while milk free triiodothyronine (fT3) concentrations were lower at 30, 60, 90 and 280 DIL, and significantly higher at 10 and 180 DIL than respective LA. Milk cortisol levels were lower at 60 and 280 DIL than LA. All measured milk hormones were significantly lower in CM1.5 compared to CM3.2, bulk-tank milk and LA. An exception was the LA of IGF-1, which was significantly lower than the IGF-1 content in CM1.5. Blood biochemical parameters fluctuated evenly during lactation and were within the reference range. Hormone concentrations in cow’s milk fluctuate during lactation, giving milk an important role in the context of public health.

Prevalence and spatial distribution of infectious diseases of dairy cattle in Ontario, Canada

Here we investigated the prevalence and spatial distribution of selected pathogens associated with infectious diseases of dairy cattle in Ontario, Canada. The cross-sectional study surveyed bulk tank milk for antibodies against bovine leukemia virus (BLV), Mycobacterium avium ssp. paratuberculosis (MAP), and Salmonella Dublin, and for the presence of mastitis pathogens (Staphylococcus aureus, Streptococcus agalactiae, Mycoplasma bovis). Between October 2021 and June 2022, bulk tank milk (BTM) samples were obtained from every commercial dairy farm in Ontario (n = 3,286). Samples underwent ELISA testing for presence of BLV, MAP and S. Dublin antibodies, and quantitative PCR testing for the detection of specific antigens of pathogens associated with mastitis. Bayesian models were used to estimate prevalence, and spatial analysis was carried out to identify regional clusters of high pathogen prevalence. Prevalence varied for different pathogens. BLV was widespread across dairy farms in Ontario, with an estimated prevalence of 88.3%. Prevalence of MAP, Staph. aureus and S. Dublin in Ontario dairy herds were 39.8%, 31.5% and 5.1%, respectively. The vast majority of dairy herds in Ontario were free of intramammary infections caused by Strep. agalactiae and M. bovis. Clusters of increased test positivity rates were detected for S. Dublin, MAP, and Staph. aureus, indicating potential geographic risk factors for pathogen transmission. For S. Dublin, an area of increased test positivity rates was detected in southwestern Ontario, close to the Canada-US border where most of the dairy herds in Ontario are located. Conversely, a localized cluster of positive test outcomes involving 14 subdivisions located in the southeastern region of Ontario was detected for Staph. aureus. Findings from our survey highlight the importance of the testing of aggregated samples and spatial analysis as part of disease surveillance programs and for implementing risk-based trading approaches among dairy producers.

An observational study demonstrates human-adapted Staphylococcus aureus strains have a higher frequency of antibiotic resistance compared to cattle-adapted strains isolated from dairy farms making farmstead cheese

BackgroundStaphylococcus aureus is a multi-host zoonotic pathogen causing human and livestock diseases. Dairy farms that make artisan cheese have distinctive concerns for S. aureus control. Antimicrobial-resistant (AMR) S. aureus is a public and animal health concern. There is a need to study the population structure of AMR S. aureus at the human-animal interface and understand the path of zoonotic transmission. This cross-sectional observational study aimed to assess the genetic diversity and AMR patterns of S. aureus isolated from cattle and humans on conventional and organic Vermont dairy farms that produce and sell farmstead cheese.ResultsA convenience sample of 19 dairy farms in Vermont was enrolled, and 160 S. aureus isolates were collected from cow quarter milk (CQM), bulk tank milk (BTM), human-hand and -nasal swabs. After deduplication, 89 isolates were used for the analysis. Sequence types (STs) were determined by multilocus sequence typing and cataloged to the PubMLST database. Nine defined and five novel STs were identified. For BTM and CQM samples, six STs were identified within cow-adapted CC97 and CC151. Two human-adapted STs were isolated from BTM and CQM. Seven human-adapted clonal complexes with eight STs were identified from human samples. One cow-adapted ST was isolated from a human. Antimicrobial susceptibility of the isolates was tested using disc diffusion and broth microdilution methods. Approximately 27% of the isolates were beta-lactam resistant and blaZ gene-positive. S. aureus isolates from human swabs were more likely to carry blaZ compared to isolates from CQM or BTM. S. aureus isolated from cows and humans on the same farm belonged to different STs.ConclusionHumans were more likely to carry beta-lactam-resistant S. aureus compared to cows, and on organic farms only human-adapted blaZ positive STs were isolated from BTM. Moreover, we identified potential spillover events of S. aureus sequence types between host species. The presence of penicillin-resistant-human-adapted S. aureus on both organic and conventional dairy farms highlights a “One Health” concern at the junction of public and animal health requiring further surveillance.

Economic losses caused by mastitis and the influence of climate variation on the occurrence of the disease in a dairy cattle farm in southern Brazil.

This study evaluated the economic impacts caused by mastitis in a small dairy farm with similar characteristics and production to most dairy farms in southern Brazil and investigated if climatic variations influenced mastitis occurrence in the region. A farm with, on average, 45 lactating Holstein cattle was monitored from November 2021 to October 2022, and data on mastitis cases, bulk tank milk somatic cell count, animal treatment costs, milk production, animal disposal costs, and production losses were collected. Monthly averages of temperature, relative humidity (RH), and rainfall in the region were obtained. The greatest loss was related to the drop in milk production, resulting in 63.8% of total losses, followed by animal disposal (29.5%), milk disposal (4.6%), and treating animals with mastitis (2.0%), totaling a 10.6% reduction in the annual gross income. There were negative correlations between the clinical mastitis rate and monthly RH and between subclinical mastitis and temperature; the occurrence of subclinical mastitis and average RH were positively correlated. Our findings showed that mastitis negatively impacted the economy and that climate influenced mastitis occurrence.

The prevalence, enterotoxigenic properties and antimicrobial susceptibility of Staphylococcus aureus isolated from various foods of animal origin

This study aimed to determine the prevalence, enterotoxigenic properties and antimicrobial resistance profile of Staphylococcus aureus isolated from 850 food samples, including bulk tank milk, Tulum cheese, chicken meat and beef carcasses, in Türkiye. S. aureus contamination rates and the mean contamination levels of 86 positive samples from bulk tank milk, Tulum cheese, chicken meat and beef carcasses were 10.8% (n:49) and 3.01±0.48 log cfu/ml, 17% (n:17) and 3.08±0.42 log cfu/g, 12% (n:12) and 2.89±0.27 log cfu/g, and 4% (n:8) and 1.28±0.54 log cfu/cm2 , respectively. 39 out of 86 isolates (45.3%) had one or more enterotoxin genes (sea-see, seg-selj, sep). sed was the most common classical enterotoxin gene, whereas sei was the most common new enterotoxin gene. Antimicrobial susceptibility testing against 16 antibiotics performed by VITEK 2 showed that 61 isolates (70.9%) were resistant to various antibiotics. 32.8% of the resistant isolates were multidrug resistant (MDR) and the mean multiple antimicrobial resistance (MAR) index was 0.301. The isolates were mostly resistant to penicillin (63.9%), followed by oxacillin (24.4%), clindamycin (19.7%) and erythromycin (12.7%). mecA was detected in 13 isolates (15.1%), but no mecC was found. It was concluded that most of the S. aureus isolates had enterotoxin genes which might cause foodborne intoxications. The high antibiotic resistance rates observed in these strains, including MRSA, may also result in some public health hazards.

Stress related to wild canid predators near dairy sheep farms associated with increased somatic cell counts in bulk-tank milk

We investigated the association between wild canid predators reported near sheep farms throughout Greece and somatic cell counts in bulk-tank milk as a reflection of milk quality. The study included 325 dairy sheep flocks, where bulk-tank milk somatic cell counts and total bacterial counts were measured and staphylococci were isolated. Farms were divided into three groups: Cohort A (farms with no reports of wild canid predators nearby), B (farms with canid predators (golden jackal and grey wolf) nearby yet with no experience of livestock losses to predation) and C (farms with canid predators nearby and livestock losses to predation). Somatic cell counts in bulk-tank milk of Cohort C farms were significantly higher, + 43% and + 29%, compared to those for Cohorts A and B, respectively: 0.617 × 106 cells mL−1 versus 0.433 × 106 or 0.477 × 106 cells mL−1, respectively. The presence of wild canid predators near sheep farms was associated with lower quality milk potentially indicative of stress consistent with the potential effects of a landscape of fear. Increasing biosecurity measures at livestock farms, e.g., fencing, and presence of livestock guard dogs could minimise predation risk, whilst also improving livestock welfare by reducing predator-associated stress.

Pathogens Associated with Bovine Mastitis: The Experience of Bosnia and Herzegovina.

To obtain improved insights into the complex microbial aetiology of bovine mastitis, this study investigated the pathogens involved in cattle mastitis in Bosnia and Herzegovina. A total of 179 milk samples from cows with clinical mastitis (CM) and subclinical mastitis (SCM), as well as eight bulk tank milk (BTM) samples from 48 dairy farms, were analysed by standard bacteriological and mycological methods. Mycoplasma detection and identification were performed using culture techniques and real-time polymerase chain reaction (PCR). A total of 88 (49.2%) mastitis samples were positive for known mastitis pathogens at 32 of 47 farms (68.1%). Mycoplasma bovis was a predominant pathogen (25/187; 13.4%) in the majority of herds (14/48; 29.2%) and accounted for 48.9% of positive CM samples. Escherichia coli was the second most dominant CM pathogen (34%), followed by Streptococcus agalactiae (10.6%), whereas Staphylococcus aureus and coagulase-negative staphylococci were the most common in SCM samples (17.1%). Other mastitis pathogens included Candida spp. and Prototheca zopfii. Two BTM samples were positive for M. bovis only, and one was positive for a mixed culture of S. aureus and Streptococcus uberis. The finding of various causative agents of bovine mastitis, with M. bovis emerging as the main pathogen, emphasizes the significance of comprehensive testing that includes not only common mastitis pathogens but also mycoplasmas, fungi, and algae.

Prevalence, Risk Factors, and Relationship between Reproductive Performance and the Presence of Antibodies against Coxiellosis in Dairy Farm Milk Tanks in the Northwest of Spain

Q fever is a zoonotic disease that has been associated with reproductive problems in animals. As there is little epidemiological data regarding the distribution and risk factors of this disorder in cattle, the objective of this study was to evaluate the prevalence of Coxiella burnetii among dairy herds in the northwest of Spain, and to determine the on-farm risk factors associated with the disease and its effects on reproductive performance. Bulk tank milk (BTM) samples were collected from 262 commercial dairy herds from A Coruña, Lugo, and Pontevedra provinces. Data about location, mean age, and herd management features were obtained. A commercial indirect ELISA kit was used to determine the presence of antibodies against C. burnetii in BTM samples. The relationship between seropositivity to C. burnetii and the risk factors was checked using a Pearson’s χ2 test and a classification tree analysis. In addition, a one-way ANOVA test and the Mann–Whitney U test were used to check the impact of seropositivity to C. burnetii on reproductive performance. A total of 60.1% of the farms tested positive for coxiellosis, the herd size, the external purchase of livestock, and the geographical area were identified as the main risk factors. Conception rate and first-service conception rate were significantly lower (p &lt; 0.05) in positive farms (37.1 and 32.9%) compared to negative farms (39.8 and 36.1%). Similarly, positive farms had significant higher incidence of endometritis (13.7% vs. 11.2%, p &lt; 0.05). Consequently, a high seropositivity and slightly negative effects of coxiellosis on reproductive performance were observed, which intensifies the need for further research, including the identification an active infection in positive herds and the characterization of the genotype.

Detection of Coxiella burnetii in the mammary gland of a dairy goat

The zoonotic bacterium Coxiella (C.) burnetii can be excreted by infected goats through birth products and milk. The detection of C. burnetii DNA in the mammary gland tissue of infected dairy goats and intermittent milk shedders has been reported, but confirmation of C. burnetii bacteria in the udder remained pending. The pathogen caused abortions in a 152-head dairy goat herd, resulting in the vaccination against C. burnetii of the entire herd with annual boosters. To monitor the C. burnetii shedding at herd level, monthly bulk tank milk (BTM) samples were analyzed using PCR (IS1111). Despite vaccination, C. burnetii DNA was detected in BTM samples within the first 16 months of the study. Therefore, individual milk samples were tested on four different occasions several months apart to identify potential intermittent milk shedders. Only one goat (#67455) tested positive three times. This goat was necropsied to investigate the presence of C. burnetii in the udder and other organs. PCR detected C. burnetii DNA solely in both mammary glands and the left teat cistern. Immunohistological examination identified C. burnetii antigen in mammary gland tissue, confirmed by the detection of C. burnetii bacteria in the mammary epithelial cells using fluorescence in situ hybridization. The removal of goat #67455 led to negative BTM samples until the end of the study. The findings demonstrate the occurrence of C. burnetii in the mammary gland of a naturally infected and vaccinated goat. The presence possibly contributed to intermittent milk shedding of goat #67455, and the mammary gland tissue may serve as a replicative niche for C. burnetii.

Phenotypic and genotypic assessment of iron acquisition in diverse bovine-associated non-aureus staphylococcal strains

Although the role of iron in bacterial infections has been well described for Staphylococcus (S.) aureus, iron acquisition in (bovine-associated) non-aureus staphylococci and mammaliicocci (NASM) remains insufficiently mapped. This study aimed at elucidating differences between four diverse bovine NASM field strains from two species, namely S. chromogenes and S. equorum, in regards to iron uptake (with ferritin and lactoferrin as an iron source) and siderophore production (staphyloferrin A and staphyloferrin B) by investigating the relationship between the genetic basis of iron acquisition through whole genome sequencing (WGS) with their observed phenotypic behavior. The four field strains were isolated in a previous study from composite cow milk (CCM) and bulk tank milk (BTM) in a Flemish dairy herd. Additionally, two well-studied S. chromogenes isolates originating from a persistent intramammary infection and from a teat apex were included for comparative purpose in all assays. Significant differences between species and strains were identified. In our phenotypical iron acquisition assay, while lactoferrin had no effect on growth recovery for all strains in iron deficient media, we found that ferritin served as an effective source for growth recovery in iron-deficient media for S. chromogenes CCM and BTM strains. This finding was further corroborated by analyzing potential ferritin iron acquisition genes using whole-genome sequencing data, which showed that all S. chromogenes strains contained hits for all three proposed ferritin reductive pathway genes. Furthermore, a qualitative assay indicated siderophore production by all strains, except for S. equorum. This lack of siderophore production in S. equorum was supported by a quantitative assay, which revealed significantly lower or negligible siderophore amounts compared to S. aureus and S. chromogenes. The WGS analysis showed that all tested strains, except for S. equorum, possessed complete staphyloferrin A (SA)-synthesis and export operons, which likely explains the phenotypic absence of siderophore production in S. equorum strains. While analyzing the staphyloferrin A and staphyloferrin B operon landscapes for all strains, we noticed some differences in the proteins responsible for iron acquisition between different species. However, within strains of the same species, the siderophore-related proteins remained conserved. Our findings contribute valuable insights into the genetic elements associated with bovine NASM pathogenesis.