Genome Comparison and Phylogenetic Analysis of Mastitis-Related Staphylococci with a Focus on Adhesion, Biofilm, and Regulatory-Related Genes

Abstract Bovine mastitis is the costliest diseases on dairy farms and is caused by different Staphylococcus species. However, staphylococci associated with clinical mastitis infections are different from subclinical ones, indicating a complex mechanism related to bovine mastitis pathogenesis. Here, we performed genomic analyses to determine the prevalence of adhesion, biofilm, and regulatory genes in 478 staphylococcal spp. associated with clinical and subclinical mastitis deposited in public databases. The most prevalent adhesin genes were the ebpS, atl, pls, sasH and sasF genes found in both clinical and subclinical isolates. However, the ebpS gene is absent in subclinical isolates of Staphylococcus arlettae, S. succinus, S. sciuri, S. equorun, S. galinarum, and S. saprophyticus. In constrast, the coa, eap, emp, efb, and vWbp genes were present more frequently in clinical mastitis isolates and highly correlated with the presence of the icaABCD and icaR biofilm genes. We also revealed that many adhesins, biofilm, and associated regulatory genes were potentially horizontally disseminated between clinical and subclinical isolates. Taken together, our results indicate that several adhesins, biofilm, and regulatory-related genes have been overlooked in previous studies and that these virulence factors may arise in staphylococcal species not generally associated with clinical mastitis by horizontal gene transfer.

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Genomic comparisons and phylogenetic analysis of mastitis-related staphylococci with a focus on adhesion, biofilm, and related regulatory genes

Scientific Reports ◽

10.1038/s41598-021-96842-2 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Lucas José Luduverio Pizauro ◽

Camila Chioda de Almeida ◽

Saura Rodrigues Silva ◽

Janet I. MacInnes ◽

Andrew M. Kropinski ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Clinical Outcomes ◽

Vaccine Development ◽

Phylogenetic Analyses ◽

Transcriptional Regulator ◽

Dairy Farms ◽

Subclinical Mastitis ◽

Regulatory Genes ◽

Genomic Analyses ◽

Highly Correlated

AbstractMastitis is a common and costly disease on dairy farms, commonly caused by Staphylococcus spp. though the various species are associated with different clinical outcomes. In the current study, we performed genomic analyses to determine the prevalence of adhesion, biofilm, and related regulatory genes in 478 staphylococcal species isolated from clinical and subclinical mastitis cases deposited in public databases. The most prevalent adhesin genes (ebpS, atl, pls, sasH and sasF) were found in both clinical and subclinical isolates. However, the ebpS gene was absent in subclinical isolates of Staphylococcus arlettae, S. succinus, S. sciuri, S. equorun, S. galinarum, and S. saprophyticus. In contrast, the coa, eap, emp, efb, and vWbp genes were present more frequently in clinical (vs. subclincal) mastitis isolates and were highly correlated with the presence of the biofim operon (icaABCD) and its transcriptional regulator, icaR. Co-phylogenetic analyses suggested that many of these adhesins, biofilm, and associated regulatory genes could have been horizontally disseminated between clinical and subclinical isolates. Our results further suggest that several adhesins, biofilm, and related regulatory genes, which have been overlooked in previous studies, may be of use for virulence profiling of mastitis-related Staphylococcus strains or as potential targets for vaccine development.

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Prevalence, risk factors and bacterial causes of bovine mastitis in southern Ethiopia

Ethiopian Veterinary Journal ◽

10.4314/evj.v24i1.4 ◽

2020 ◽

Vol 24 (1) ◽

Author(s):

Rahmeto Abebe ◽

Mesele Abera ◽

Yifat Denbarga ◽

Mishamo Suleyman ◽

Amene Fekadu ◽

...

Keyword(s):

Risk Factors ◽

Bovine Mastitis ◽

Dairy Farms ◽

Subclinical Mastitis ◽

Clinical Mastitis ◽

Southern Ethiopia ◽

Cross Sectional ◽

Microbiological Methods ◽

Associated Risk Factors ◽

And Control

Mastitis is one of the most economically important diseases in dairy farms worldwide. It is particularly important in Ethiopia where no routine prevention and control practices are in place. This cross-sectional study was carried out between October 2017 and June 2018 to estimate the prevalence of mastitis, identify the associated risk factors and isolate bacterial causes in dairy farms located in southern Ethiopia using standard microbiological methods and questionnaire survey. A total of 686 lactating cows which were found in 122 selected dairy farms were investigated by physical examination and California mastitis test. The overall cow-level prevalence of mastitis was 54.2% (95% CI: 50.5 – 57.9%). Based on the study site, the prevalence was 55.7% in Hawassa, 54.3% in Arsi Negele, 52.6% each in Wondo Genet and Wolayta Soddo towns with no significant (p > 0.05) difference among the sites. The majority of mastitis cases were subclinical (48.1%) while the clinical mastitis was only 6.1%. Of the 122 herds tested, 109 (89.3%; 95% CI: 82.1 – 93.9%) had at least a cow positive for mastitis. The study showed that high parity number (OR = 1.6; p = 0.015), flat (OR = 4.5; p <0.001) and round (OR = 2; p <0.001) teat end shape, history of mastitis in preceding lactation (OR = 3.3; p <0.001), and slightly (OR = 3.5; p < 0.001), moderately (OR = 4.9; p < 0.001), and very dirty (OR = 9.2; p < 0.001) udder and legs were the major risk factors which are significantly associated with higher prevalence of mastitis. Based on the available media and reagents, the major bacteria isolated from subclinical mastitic milk samples were Staphylococcus spp. (57.3%), Streptococcus spp. (18.6%), E. coli (17.3%) and Bacillus spp. (7.5%) in order of their abundance. The present study revealed a high prevalence of mastitis, particularly the subclinical one, and the associated risk factors. Enhancing the awareness of dairy farmers, regular screening of cows for subclinical mastitis, proper treatment of the clinical cases, improving the hygienic condition of the cows, and culling of chronically infected cows are critically important to prevent and control bovine mastitis. Keywords: Mastitis, Prevalence, Risk factors, Southern Ethiopia

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Economic Impacts of Clinical and Sub Clinical Mastitis on Dairy Farms

Veterinary Science Research ◽

10.30564/vsr.v3i2.4119 ◽

2021 ◽

Vol 3 (2) ◽

Author(s):

Amjad Islam Aqib ◽

Afshan Muneer ◽

Muhammad Shafeeq ◽

Nimra Kirn

Keyword(s):

Milk Production ◽

Economic Impacts ◽

Bovine Mastitis ◽

Dairy Farms ◽

Economic Loss ◽

Subclinical Mastitis ◽

Milk Composition ◽

Clinical Mastitis ◽

Economic Losses ◽

Invasive Infection

Studies have reported on the economic impacts of clinical and subclinical mastitis on dairy farms. Bovine mastitis is a disorder that affects dairy farms and has a major economic impact. Most of the economic losses are the result of mastitis. Mastitis is an invasive infection that is among the most numerous and highly complicated infections in the dairy sector. Mastitis is one of the most expensive diseases in terms of production losses among animal diseases. Mastitis reduces milk production, changes milk composition, and shortens the productive life of infected cows. Farmers must concentrate on avoiding mastitis infection whilst putting in place and following a mastitis control programed. Bovine mastitis, the most significant disease of dairy herds, has huge effects on farm economics. Mastitis losses are due to reduced milk production, the cost of treatments, and culling. Major factors related to low milk yield could be low genetic potential as well as poor nutritional and managerial approaches. Most of the losses are related to somatic cell count (SCC), which is characterised by an increase in the percentage of milk. Culling costs are the costs of rearing or buying a replacement animal, mostly heifers. Overhead impacts include the replacement animals' lower milk supply effectiveness. The expense of replacing animals prematurely due to mastitis is one of the most significant areas of economic loss.

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Alternative treatment of bovine mastitis

Veterinarska stanica ◽

10.46419/vs.52.6.9 ◽

2021 ◽

Vol 52 (6) ◽

Author(s):

Nora Mimoune ◽

Radhwane Saidi ◽

Oumayma Benadjel ◽

Djamel Khelef ◽

Rachid Kaidi

Keyword(s):

Bovine Mastitis ◽

Dairy Farms ◽

Subclinical Mastitis ◽

Tolerance Test ◽

Clinical Mastitis ◽

Cure Rate ◽

Good Tolerance ◽

Milk Samples ◽

Cull Cows ◽

Homeopathic Preparation

The treatment of mastitis is based mainly on the use of antibiotics. However, in recent times, an increase in resistance phenomena and the presence of residues in milk and their derivatives has been reported. Few studies have focused on the treatment of mastitis by homeopathy elsewhere or in Algeria. The objective of this current study aimed to clarify in particular the interest of homeopathy in the treatment of mastitis. The study was carried out on two dairy farms on a total of 14 cows located in Laghouat region (southern Algeria). Before the start of the experiment, a tolerance test was performed on two cull cows free from any apparent infection (except mastitis). Local and general reactions were noted at specific times. A total of 31 mammary quarters received 4 intramammary injections of a homeopathic preparation containing several natural products every 12h over 48h. A clinical examination and an analysis of the milk samples on D0, D7 and D14 were performed on all cows and the conclusion was made on the day 14. Data showed a very good tolerance to the homeopathic preparation and a 75% cure rate of the clinical mastitis. An improvement with a decrease in CMT score was also noted. In addition, healing rates of 51.85% for subclinical mastitis on D7 and 59.29% on D14 were also reported. Ultimately, homeopathy could, in some cases, represent an alternative to antibiotic therapy and bring an advantage to breeders. Further investigations should be performed in the future.

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Variations in the levels of acute-phase proteins and lactoferrin in serum and milk during bovine subclinical mastitis

Journal of Dairy Research ◽

10.1017/s002202992100056x ◽

2021 ◽

pp. 1-5

Author(s):

Aarsha Raj ◽

Vinodkumar Kulangara ◽

Tresamol P. Vareed ◽

Deepa P. Melepat ◽

Latha Chattothayil ◽

...

Keyword(s):

Acute Phase ◽

Acute Phase Proteins ◽

Solid Phase ◽

Bovine Mastitis ◽

Subclinical Mastitis ◽

Clinical Mastitis ◽

Reactive Protein ◽

Amyloid A ◽

California Mastitis Test ◽

Bacterial Organism

Abstract Variations in the levels of acute phase proteins and lactoferrin in serum and milk for diagnosis of subclinical mastitis in dairy cows are described in this research paper. Milking animals from two organized dairy farms in Kerala, India, were screened by California Mastitis Test (CMT), Electrical Conductivity test (EC) and Somatic Cell Count (SCC) test to identify animals affected with sub clinical mastitis (SCM). The concentrations of acute phase proteins (APP) Haptoglobin (Hp), C- reactive protein (CRP), Albumin, Lactoferrin (Lf) and α- 1 acid glycoprotein (AGP) in milk and Hp, Albumin, Serum Amyloid A (SAA) and CRP in the serum of 40 normal cows and 40 cows affected with sub clinical mastitis were assessed. Solid phase ELISA was employed for assessment of all parameters except the albumin levels, for which spectrophotometry was used. The values of Hp in milk; and SAA, AGP and Lf in serum, were significantly elevated in the group with sub clinical mastitis. Such variations were found to be independent of the specific bacterial organism causing the disease. These results show that significant variations exist in the levels of acute phase proteins Hp, AGP and Lf in milk, and SAA in serum of animals affected with subclinical bovine mastitis that are not affected by specific bacterial etiology.

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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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Characterization of Staphylococcus aureus Isolates From Cases of Clinical Bovine Mastitis on Large-Scale Chinese Dairy Farms

Frontiers in Veterinary Science ◽

10.3389/fvets.2020.580129 ◽

2020 ◽

Vol 7 ◽

Author(s):

Kangjun Liu ◽

Luyao Tao ◽

Jianji Li ◽

Li Fang ◽

Luying Cui ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Cluster Analysis ◽

Antimicrobial Resistance ◽

Large Scale ◽

Bovine Mastitis ◽

Dairy Farms ◽

Clinical Mastitis ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Tof Ms

Bovine mastitis is a prevalent disease that causes serious economic problems globally in the dairy industry. Staphylococcus aureus is an important pathogen of bovine mastitis. This study was conducted to characterize S. aureus isolates from clinical bovine mastitis cases in large-scale dairy herds in China. S. aureus was isolated from 624 clinical mastitis cases and confirmed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). In total, 62 S. aureus isolates were obtained. Cluster analysis, genetic diversity, quantification of biofilm formation, antimicrobial resistance, and detection of virulence genes were performed on these isolates of S. aureus. Eight isolates harbored the mecA gene and were sensitive to oxacillin. MALDI-TOF MS cluster analysis revealed that the 62 isolates were divided into three major clusters (I, II, III) and eight main groups (A–H) at the distance level of 700. The agr II was the most prevalent (56.5%). The 62 S. aureus isolates were assigned to seven spa types. The most common spa type was t529(58.1%), followed by t2196 (14.5%), t518 (14.5%), t571(6.5%), t034 (3.2%), t2734 (1.6%), and t730 (1.6%). Five STs were identified from seven representative isolates as follows: ST630/CC8, ST97/CC97, ST50, ST398, and ST705. All isolates had the ability to form biofilm. Antimicrobial resistance was most frequently observed to ciprofloxacin (29%), followed by penicillin (24.2%), and streptomycin (9.6%). All isolates harbored the fnbA, clfB (100%), icaA, and icaD genes. This study provides the basis for the development of bovine mastitis prevention program on large-scale dairy farms.

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Prevalence of clinical and subclinical mastitis and quality of milk on smallholder dairy farms in Tanzania

Journal of the South African Veterinary Association ◽

10.4102/jsava.v80i3.195 ◽

2009 ◽

Vol 80 (3) ◽

Cited By ~ 14

Author(s):

R.H. Mdegela ◽

R. Ryoba ◽

E.D. Karimuribo ◽

E.J. Phiri ◽

T. Loken ◽

...

Keyword(s):

Antimicrobial Agents ◽

Dairy Farms ◽

Subclinical Mastitis ◽

Clinical Mastitis ◽

Milk Quality ◽

Herd Level ◽

Milk Samples ◽

Antimicrobial Residues ◽

Smallholder Dairy ◽

Smallholder Dairy Farms

A cross sectional study was conducted during October and November 2006 on 69 smallholder dairy farms with lactating cows in Mvomero and Njombe districts Tanzania, to determine the prevalence of mastitis and to assess the milk quality on the study farms. Clinical mastitis was investigated using clinical changes of udder and milk at animal level. Cow-side California Mastitis Test (CMT) and microbiological cultures were used to assess subclinical mastitis at quarter level. Milk quality was determined on bulk milk samples at herd level using alcohol and acidity tests, butter fat content, total solids, ash content as well as Delvotest® for antimicrobial residues. Overall prevalence of clinical mastitis at herd level in both districts was 21.7 % (n = 69). Based on CMT, prevalence of subclinical mastitis at animal level was 51.6 % (n = 91). Prevalence of bacterial isolates at animal level was 35.2 % (n = 91) while for fungal it was 16.7 % (n = 90). Based on CMT results, prevalence of subclinical mastitis at quarter level was 30 % (n = 353), while for bacteria and fungi it was 16 % and 6 % respectively. Contamination of milk with antimicrobial residues was 4.5 % (n =67). The milk quality parameters for most of the milk samples were within acceptable levels. Findings in this study have demonstrated high prevalence of subclinical mastitis that may contribute to low productivity of dairy cattle in both districts. About 20 % of CMT subclinical cases had no involvement of microbial pathogens that suggested the need for minimal interventions with antimicrobial agents. These findings call for use of udder disinfectants and improved milking hygiene as intervention strategies to control mastitis on the smallholder dairy farms in Tanzania.

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Molecular Detection and Sensitivity to Antibiotics and Bacteriocins of Pathogens Isolated from Bovine Mastitis in Family Dairy Herds of Central Mexico

BioMed Research International ◽

10.1155/2015/615153 ◽

2015 ◽

Vol 2015 ◽

pp. 1-9 ◽

Cited By ~ 12

Author(s):

Ma. Fabiola León-Galván ◽

José E. Barboza-Corona ◽

A. Arianna Lechuga-Arana ◽

Mauricio Valencia-Posadas ◽

Daniel D. Aguayo ◽

...

Keyword(s):

Bacillus Thuringiensis ◽

Bovine Mastitis ◽

Dairy Herd ◽

Subclinical Mastitis ◽

Herd Size ◽

Clinical Mastitis ◽

Dairy Herds ◽

Coagulase Negative Staphylococci ◽

California Mastitis Test ◽

Streptococcus Uberis

Thirty-two farms (n=535cows) located in the state of Guanajuato, Mexico, were sampled. Pathogens from bovine subclinical mastitis (SCM) and clinical mastitis (CLM) were identified by 16S rDNA and the sensitivity to both antibiotics and bacteriocins ofBacillus thuringiensiswas tested. Forty-six milk samples were selected for their positive California Mastitis Test (CMT) (≥3) and any abnormality in the udder or milk. The frequency of SCM and CLM was 39.1% and 9.3%, respectively. Averages for test day milk yield (MY), lactation number (LN), herd size (HS), and number of days in milk (DM) were 20.6 kg, 2.8 lactations, 16.7 animals, and 164.1 days, respectively. MY was dependent on dairy herd (DH), LN, HS, and DMP<0.01, and correlations between udder quarters from the CMT were around 0.49P<0.01. Coagulase-negative staphylococci were mainly identified, as well asStaphylococcus aureus,Streptococcus uberis,Brevibacterium stationis,B. conglomeratum, andStaphylococcus agnetis. Bacterial isolates were resistant to penicillin, clindamycin, ampicillin, and cefotaxime. Bacteriocins synthesized byBacillus thuringiensisinhibited the growth of multiantibiotic resistance bacteria such asS. agnetis,S. equorum,Streptococcus uberis,Brevibacterium stationis, andBrachybacterium conglomeratum, but they were not active againstS. sciuri, a microorganism that showed an 84% resistance to antibiotics tested in this study.

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N-acetyl -β-D-glucosaminidase activity in cow milk as an indicator of mastitis

Journal of Dairy Research ◽

10.1017/s0022029916000224 ◽

2016 ◽

Vol 83 (2) ◽

pp. 219-227 ◽

Cited By ~ 19

Author(s):

Mari Hovinen ◽

Heli Simojoki ◽

Reeta Pösö ◽

Jenni Suolaniemi ◽

Piret Kalmus ◽

...

Keyword(s):

Somatic Cell Count ◽

Bovine Mastitis ◽

Reference Value ◽

Subclinical Mastitis ◽

Clinical Mastitis ◽

Cow Milk ◽

Milk Samples ◽

High Level ◽

Accurate Indicator

Activity of lysosomal N-acetyl-β-d-glucosaminidase (NAGase) in milk has been used as an indicator of bovine mastitis. We studied NAGase activity of 808 milk samples from healthy quarters and quarters of cows with spontaneous subclinical and clinical mastitis. Associations between milk NAGase activity and milk somatic cell count (SCC), mastitis causing pathogen, quarter, parity, days in milk (DIM) and season were studied. In addition, the performance of NAGase activity in detecting clinical and subclinical mastitis and distinguishing infections caused by minor and major bacteria was investigated. Our results indicate that NAGase activity can be used to detect both subclinical and clinical mastitis with a high level of accuracy (0·85 and 0·99). Incomplete correlation between NAGase activity and SCC suggests that a substantial proportion of NAGase activity comes from damaged epithelial cells of the udder in addition to somatic cells. We therefore recommend determination of NAGase activity from quarter foremilk after at least six hours from the last milking using the method described. Samples should be frozen before analysis. NAGase activity should be interpreted according to DIM, at least during the first month of lactation. Based on the results of the present study, a reference value for normal milk NAGase activity of 0·1–1·04 pmoles 4-MU/min/μl for cows with ≥30 DIM (196 samples) could be proposed. We consider milk NAGase activity to be an accurate indicator of subclinical and clinical mastitis.

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