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21 pages, 1503 KiB

Open AccessArticle

Transcriptomic Analysis of Circulating Leukocytes Obtained during the Recovery from Clinical Mastitis Caused by Escherichia coli in Holstein Dairy Cows

by Zhangrui Cheng, Sergio Palma-Vera, Laura Buggiotti, Mazdak Salavati, Frank Becker, Dirk Werling, D. Claire Wathes and GplusE Consortium

Animals 2022, 12(16), 2146; https://doi.org/10.3390/ani12162146 - 21 Aug 2022

Cited by 7 | Viewed by 2510

Abstract

The risk and severity of clinical infection with Escherichia coli as a causative pathogen for bovine mastitis is influenced by the hosts’ phenotypic and genotypic variables. We used RNA-Seq analysis of circulating leukocytes to investigate global transcriptomic profiles and genetic variants from Holstein [...] Read more.

The risk and severity of clinical infection with Escherichia coli as a causative pathogen for bovine mastitis is influenced by the hosts’ phenotypic and genotypic variables. We used RNA-Seq analysis of circulating leukocytes to investigate global transcriptomic profiles and genetic variants from Holstein cows with naturally occurring cases of clinical mastitis, diagnosed using clinical symptoms and milk microbiology. Healthy lactation-matched cows served as controls (CONT, n = 6). Blood samples were collected at two time periods during the recovery phase post diagnosis: EARLY (10.3 ± 1.8 days, n = 6) and LATE (46.7 ± 11 days, n = 3). Differentially expressed genes (DEGs) between the groups were identified using CLC Genomics Workbench V21 and subjected to enrichment analysis. Variant calling was performed following GATKv3.8 best practice. The comparison of E. coli(+) EARLY and CONT cows found the up-regulation of 1090 DEGs, mainly with immune and inflammatory functions. The key signalling pathways involved NOD-like and interleukin-1 receptors and chemokines. Many up-regulated DEGs encoded antimicrobial peptides including cathelicidins, beta-defensins, S100 calcium binding proteins, haptoglobin and lactoferrin. Inflammation had largely resolved in the E. coli(+) LATE group, with only 29 up-regulated DEGs. Both EARLY and LATE cows had up-regulated DEGs encoding ATP binding cassette (ABC) transporters and haemoglobin subunits were also up-regulated in LATE cows. Twelve candidate genetic variants were identified in DEGs between the infected and CONT cows. Three were in contiguous genes WIPI1, ARSG and SLC16A6 on BTA19. Two others (RAC2 and ARHGAP26) encode a Rho-family GTPase and Rho GTPase-activating protein 26. These results show that the initial inflammatory response to E. coli continued for at least 10 days despite prompt treatment and provide preliminary evidence for genetic differences between cows that may predispose them to infection. Full article

(This article belongs to the Special Issue Clinical Mastitis and Intramammary Infections in Dairy Farms)

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18 pages, 1133 KiB

Open AccessArticle

Dairy Cows’ Udder Pathogens and Occurrence of Virulence Factors in Staphylococci

by František Zigo, Zuzana Farkašová, Jana Výrostková, Ivana Regecová, Silvia Ondrašovičová, Mária Vargová, Naďa Sasáková, Ewa Pecka-Kielb, Šárka Bursová and David Sandor Kiss

Animals 2022, 12(4), 470; https://doi.org/10.3390/ani12040470 - 14 Feb 2022

Cited by 23 | Viewed by 2745

Abstract

This study investigated 960 Slovak and Czech spotted cattle from four different conventional (non-organic) dairy herds located in Eastern Slovakia and Czechia during early lactation (14–100 days after calving). Dairy cows were examined clinically; milk from fore-stripping of each udder quarter was subjected [...] Read more.

This study investigated 960 Slovak and Czech spotted cattle from four different conventional (non-organic) dairy herds located in Eastern Slovakia and Czechia during early lactation (14–100 days after calving). Dairy cows were examined clinically; milk from fore-stripping of each udder quarter was subjected to sensory examination and assessed by the California mastitis test (CMT), and laboratory analyses of bacterial pathogens in milk, including virulence factors, were conducted. Positive CMT scores (1–3) for one or more quarters were detected in 271 (28.2%) of the examined animals. Out of 230 infected milk samples, representing 24.0% of all dairy cows, staphylococci (59.1% of positive findings) were the most commonly isolated organisms, followed by E. coli (11.3%), streptococci Str. uberis (9.1%) and Str. agalactiae (3.4%), and enterococci (6.1%). From 136 isolates of S. aureus (38 isolates) and non-aureus staphylococci (NAS; 98 isolates), virulence factors and their resistance to 14 antimicrobials were detected using the disk diffusion method, with PCR detection of the methicillin resistance gene, mecA. An increased incidence of clinical and chronic forms of mastitis has been reported in mastitic cows in which staphylococci, especially S. aureus and NAS (S. chromogenes, S. warneri, and S. xylosus), have been detected and compared to other isolated udder pathogens. From those species, S. aureus and isolates of NAS mentioned above showed multiple virulence factors that are more likely to hydrolyze DNA, hemolysis, produce gelatinase and biofilm, and have multi-drug resistance as compared to other less virulent staphylococci. Generally, the isolated staphylococci showed 77.2% resistance to one or more antimicrobials, in particular to aminoglycosides, β-lactams, macrolides, or cephalosporins. Isolates that showed the ability to form a biofilm were more resistant to more than one antimicrobial than isolates without biofilm production. Multi-drug resistance to three or more antimicrobial classes was recorded in 16 isolates (11.7%), and the presence of the mecA gene was also confirmed in two isolates of S. aureus and two species of NAS. Full article

(This article belongs to the Special Issue Clinical Mastitis and Intramammary Infections in Dairy Farms)

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18 pages, 3162 KiB

Open AccessArticle

Investigations on Transfer of Pathogens between Foster Cows and Calves during the Suckling Period

by Katharina Köllmann, Nicole Wente, Yanchao Zhang and Volker Krömker

Animals 2021, 11(9), 2738; https://doi.org/10.3390/ani11092738 - 19 Sep 2021

Cited by 1 | Viewed by 2637

Abstract

To date, there have been few studies on the health effects of foster cow systems, including the transmission of mastitis-associated pathogens during suckling. The present study aimed to compare the pathogens detected in the mammary glands of the foster cow with those in [...] Read more.

To date, there have been few studies on the health effects of foster cow systems, including the transmission of mastitis-associated pathogens during suckling. The present study aimed to compare the pathogens detected in the mammary glands of the foster cow with those in the oral cavities of the associated foster calves and to evaluate the resulting consequences for udder health, calf health and internal biosecurity. Quarter milk sampling of 99 foster cows from an organic dairy farm was conducted twice during the foster period. Oral cavity swabs were taken from 345 foster calves. Furthermore, quarter milk samples were collected from 124 biological dams to investigate possible transmission to the foster cows via the suckling calves. All samples were microbiologically examined and confirmed by MALDI-TOF (matrix-assisted laser desorption time-of-flight mass-spectrometry). Using RAPD-PCR (randomly amplified polymorphic DNA polymerase chain reaction), strain similarities were detected for Pasteurella multocida, Staphylococcus aureus, S. sciuri and Streptococcus (Sc.) suis. Transmission of P. multocida and S. aureus probably occurred during suckling. For S. sciuri and Sc. suis, environmental origins were assumed. Transmission from dam to foster cow with the suckling calf as vector could not be clearly demonstrated. Full article

(This article belongs to the Special Issue Clinical Mastitis and Intramammary Infections in Dairy Farms)

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14 pages, 670 KiB

Open AccessArticle

Effect of Pre-Milking Teat Foam Disinfection on the Prevention of New Mastitis Rates in Early Lactation

by Sarah Rose Fitzpatrick, Mary Garvey, Jim Flynn, Bernadette O’Brien and David Gleeson

Animals 2021, 11(9), 2582; https://doi.org/10.3390/ani11092582 - 03 Sep 2021

Cited by 2 | Viewed by 2443

Abstract

The objective of this study was to determine the benefit of pre-milking teat foam disinfection on the prevention of new infections by contagious and environmental bacteria in two spring calving herds managed outdoors (Herd 1 [H1]; 331 cows and Herd 2 [H2]; 142 [...] Read more.

The objective of this study was to determine the benefit of pre-milking teat foam disinfection on the prevention of new infections by contagious and environmental bacteria in two spring calving herds managed outdoors (Herd 1 [H1]; 331 cows and Herd 2 [H2]; 142 cows). Four pre-milking teat preparation treatments were applied post calving; with each herd receiving two treatments; using a split udder design (for approx. 15 weeks). These treatments included; (1) ‘water wash, foam application and dry wipe (WFD) in H1′; (2) ‘water wash and dry wipe (WD)’ in H1; (3) ‘foam application and dry wipe (FD)’ in H2; (4) ‘no teat cleaning preparation (NP)’ in H2. Individual quarter foremilk samples were collected on four occasions and all clinical and sub-clinical cases were recorded. The mean SCC of quarter foremilk samples was 134 × 10³ cells/mL and 127 × 10³ cells/mL for WD and WFD, respectively, and 109 × 10³ cells/mL and 89 × 10³ cells/mL for NP and FD, respectively (p > 0.05). Lower bacterial counts were observed on teat skin that received a foaming treatment. Pre-milking teat disinfection using a foaming product may be of little benefit, in early lactation, for a pasture-based dairy herd. Full article

(This article belongs to the Special Issue Clinical Mastitis and Intramammary Infections in Dairy Farms)

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17 pages, 1025 KiB

Open AccessArticle

Incidence, Etiology, and Risk Factors of Clinical Mastitis in Dairy Cows under Semi-Tropical Circumstances in Chattogram, Bangladesh

by Shuvo Singha, Gerrit Koop, Ylva Persson, Delower Hossain, Lauren Scanlon, Marjolein Derks, Md. Ahasanul Hoque and Md. Mizanur Rahman

Animals 2021, 11(8), 2255; https://doi.org/10.3390/ani11082255 - 30 Jul 2021

Cited by 9 | Viewed by 4795

Abstract

Clinical mastitis (CM) is an important production disease in dairy cows, but much of the knowledge required to effectively control CM is lacking, specifically in low-income countries where most farms are small and have specific dairy management, such as regular udder cleaning and [...] Read more.

Clinical mastitis (CM) is an important production disease in dairy cows, but much of the knowledge required to effectively control CM is lacking, specifically in low-income countries where most farms are small and have specific dairy management, such as regular udder cleaning and practicing hand milking. Therefore, we conducted a 6-month-long cohort study to (a) estimate the incidence rate of clinical mastitis (IRCM) at the cow and quarter level, (b) identify risk factors for the occurrence of CM, (c) describe the etiology of CM, and (d) quantify antimicrobial susceptibility (AMS) against commonly used antimicrobial agents in S. aureus and non-aureus Staphylococcus spp. (NAS) in dairy farms in the Chattogram region of Bangladesh. On 24 farms, all cows were monitored for CM during a 6-month period. Cases of CM were identified by trained farmers and milk samples were collected aseptically before administering any antimicrobial therapy. In total, 1383 lactating cows were enrolled, which totaled 446 cow-years at risk. During the study period, 196 new cases of CM occurred, resulting in an estimated crude IRCM of 43.9 cases per 100 cow-years, though this varied substantially between farms. Among the tested CM quarter samples, Streptococci (22.9%) followed by non-aureus staphylococci (20.3%) were the most frequently isolated pathogens and resistance of S. aureus and NAS against penicillin (2 out of 3 and 27 out of 39 isolates, respectively) and oxacillin (2 out of 3 and 38 out of 39 isolates, respectively) was common. The IRCM was associated with a high milk yield, 28 to 90 days in milk, and a higher body condition score. Our results show that there is substantial room for udder health improvement on most farms. Full article

(This article belongs to the Special Issue Clinical Mastitis and Intramammary Infections in Dairy Farms)

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17 pages, 333 KiB

Open AccessArticle

Microbial Aetiology, Antibiotic Susceptibility and Pathogen-Specific Risk Factors for Udder Pathogens from Clinical Mastitis in Dairy Cows

by Anna Duse, Karin Persson-Waller and Karl Pedersen

Animals 2021, 11(7), 2113; https://doi.org/10.3390/ani11072113 - 16 Jul 2021

Cited by 28 | Viewed by 4913

Abstract

Mastitis is one of the most important infectious diseases and one of the diseases that causes the greatest use of antibiotics in dairy cows. Therefore, updated information on the bacteria that cause mastitis and their antibiotic susceptibility properties is important. Here, for the [...] Read more.

Mastitis is one of the most important infectious diseases and one of the diseases that causes the greatest use of antibiotics in dairy cows. Therefore, updated information on the bacteria that cause mastitis and their antibiotic susceptibility properties is important. Here, for the first time in over 10 years, we updated the bacterial findings in clinical mastitis in Swedish dairy cows together with their antibiotic resistance patterns and risk factors for each bacterial species. During the period 2013–2018, samples from clinical mastitis were collected, together with information on the cows and herds of origin. The samples were cultured, and a total of 664 recovered bacterial isolates were subjected to susceptibility testing. Staphylococcus aureus (S. aureus) was the most common pathogen and accounted for 27.8% of diagnoses, followed by Streptococcus dysgalactiae (S. dysgalactiae) (15.8%), Escherichia coli (E. coli) (15.1%), Streptococcus uberis (S. uberis) (11.4%), Trueperella pyogenes (T. pyogenes) (7.7%), non-aureus staphylococci (NAS) (2.8%), Klebsiella spp. (2.7%), Enterococcus spp. (1.3%), and Streptococcus agalactiae (S. agalactiae) (1.2%). Various other bacteria accounted for 2.6%. Staphylococci were, in general, susceptible to most antibiotics, but 2.6% of S. aureus and 30.4% of NAS were resistant to penicillin. No methicillin-resistant staphylococci were found. All S. agalactiae were susceptible to penicillin. Bimodal and trimodal MIC distributions for penicillin in S. dysgalactiae and S. uberis, respectively, indicate acquired reduced susceptibility in some isolates. The mostly unimodal MIC distributions of T. pyogenes indicate that acquired resistance does usually not occur in this species. Among E. coli, 14.7% were resistant to at least one antibiotic, most often ampicillin (8.7%), streptomycin (7.8%), or sulphamethoxazole (6.9%). Klebsiella spp. had low resistance to tetracycline (9.1%) but is considered intrinsically resistant to ampicillin. Pathogen-specific risk factors were investigated using multivariable models. Staphylococcus aureus, S. dysgalactiae, and T. pyogenes were more common, while E. coli was less common in quarters with more than one pathogen. S. aureus and T. pyogenes were mostly seen in early lactation, while E. coli was more common in peak to mid lactation and S. dysgalactiae in early to peak lactation. Trueperella pyogenes and Klebsiella spp. were associated with a previous case of clinical mastitis in the current lactation. Staphylococcus aureus was associated with tie stalls and T. pyogenes with loose housing. All pathogens except E. coli and S. dysgalactiae had a seasonal distribution. In conclusion, the aetiological agents for clinical bovine mastitis have remained relatively stable over the last 10–15 years, S. aureus, S. dysgalactiae, E. coli and S. uberis being the most important. Resistance to penicillin among Gram-positive agents was low, and in general, antibiotic resistance to other compounds was low among both Gram-positive and Gram-negative agents. Full article

(This article belongs to the Special Issue Clinical Mastitis and Intramammary Infections in Dairy Farms)

10 pages, 265 KiB

Open AccessCommunication

Risk Factors Associated With Mastitis in Smallholder Dairy Farms in Southeast Brazil

by Aline Callegari Silva, Richard Laven and Nilson Roberti Benites

Animals 2021, 11(7), 2089; https://doi.org/10.3390/ani11072089 - 14 Jul 2021

Cited by 10 | Viewed by 4206

Abstract

The aim of this study was to investigate the potential risk factors for clinical and subclinical mastitis in smallholder dairy farms in Brazil. A prospective, repeated cross-sectional study was carried out between May 2018 and June 2019 on 10 smallholder dairy farms. Potential [...] Read more.

The aim of this study was to investigate the potential risk factors for clinical and subclinical mastitis in smallholder dairy farms in Brazil. A prospective, repeated cross-sectional study was carried out between May 2018 and June 2019 on 10 smallholder dairy farms. Potential risk factors for subclinical and clinical mastitis at the herd and cow level were recorded through interviewing the owner and by observation. A combination of clinical udder examination and the Tamis (screened mug with a dark base) test (Tadabras Indústria e Comércio de Produtos Agrovetereinário LTDA, Bragrança Paulista, SP, Brazil) were applied to observe clinical mastitis, and the California Mastitis Test (Tadabras Indústria e Comércio de Produtos Agrovetereinário LTDA, Bragrança Paulista, SP, Brazil) was used to determine subclinical mastitis. A total of 4567 quarters were tested, 107 (2.3%) had clinical mastitis, while 1519 (33.2%) had subclinical mastitis. At the cow level, clinical mastitis risk was highest in mid-lactation (50–150 days in milk) with OR 2.62 with 95% confidence interval (CI) of 1.03–6.67, while subclinical mastitis was highest in late lactation (> 150 days in milk) with OR 2.74 (95% CI 2.05–3.63) and lower in primiparous (OR 0.54, 95% CI 0.41–0.71) than multiparous cows. At the herd level, using dry-cow treatment (OR 4.23, 95% CI 1.42–12.62) was associated with an increased risk of clinical mastitis. Milking clinical (OR 0.37, 95% CI 0.24–0.56) and subclinical cases last (OR 0.21, 95% CI 0.09–0.47) and cleaning the milking parlor regularly (OR 0.27, 95% CI 0.15–0.46) had decreased odds for subclinical mastitis, while herds with optimized feed had greater odds (OR 9.11, 95% CI 2.59–31.9). Prevalence of clinical mastitis was at its lowest at the first visit in June/July and highest at the last visit in April/June (OR 3.81, 95% CI 1.93–7.52). Subclinical mastitis also presented increased odds in the last visit (OR 2.62, 95% CI 2.0–3.36). This study has identified some risk factors for mastitis on smallholder farms but further research on more farms across more areas of Brazil is required to develop a targeted mastitis control program for smallholder farms. Full article

(This article belongs to the Special Issue Clinical Mastitis and Intramammary Infections in Dairy Farms)

14 pages, 915 KiB

Open AccessArticle

Total and Differential Cell Counts as a Tool to Identify Intramammary Infections in Cows after Calving

by Alfonso Zecconi, Gabriele Meroni, Valerio Sora, Roberto Mattina, Micaela Cipolla and Lucio Zanini

Animals 2021, 11(3), 727; https://doi.org/10.3390/ani11030727 - 07 Mar 2021

Cited by 12 | Viewed by 1972

Abstract

Milk differential somatic cells count (DSCC), made possible under field conditions by the recent availability of a high-throughput milk analyzer may represent an improvement in mastitis diagnosis. While an increasing number of studies reports data on DSCC on individual cow samples, very few [...] Read more.

Milk differential somatic cells count (DSCC), made possible under field conditions by the recent availability of a high-throughput milk analyzer may represent an improvement in mastitis diagnosis. While an increasing number of studies reports data on DSCC on individual cow samples, very few concerns DSCC from quarter milk samples. This paper reports for the first time the results of a retrospective study aiming to assess the performance of total (SCC), DSCC, and a novel calculated marker (PLCC) measured on quarter milk samples as a method to identify cows at risk for intramammary infection (IMI) in the first 30 days after calving. Overall, 14,586 valid quarter milk samples (3658 cows) taken in the first 30 days of lactation were considered. Quarters with major pathogens (MP) IMI, as expected, showed significantly higher means for SCC, DSCC, and PLCC. The accuracy, sensitivity, and specificity of the diagnosis based on different cut-offs calculated by ROC analysis are relatively close among DSCC, PLCC, and SCC (up to cut-off of 200,000 cells/mL). However, decision-tree analysis which includes the costs of analysis, but also the costs of the actions taken after test results showed as PLCC has the lowest cost among the three markers, and PLCC and SCC are cost effective when MP prevalence is higher than 6–10%. This diagnostic approach is of high interest particularly when selective dry cow therapy is applied to improve animal health at the herd level. Full article

(This article belongs to the Special Issue Clinical Mastitis and Intramammary Infections in Dairy Farms)

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