The frequency of bacterial pathogens of mastitis and their antibiotic susceptibility in Saanen and Alpine goats

Mastitis is mammary gland inflammation and as the purpose of Saanen and Alpine farms is milk production, control of mastitis is important. Antimicrobial resistance among mastitis pathogens has gradually been increasing. The aim of the present study was to isolate pathogenic bacteria from mastitis cases in Saanen and Alpine goats and to determine their antibiotic resistance in milk. The milk sample of 26 Saanen and 29 Alpine breeds with clinical mastitis was collected and cultured on general microbiological media. Then, the colonies were stained by gram staining and were assessed by differential media and biochemical tests. PCR was performed for the detection of Mycoplasma spp. The isolated bacteria were tested against 12 antibacterial disks. The significant difference in drug resistance levels between the Saanen and Alpine breeds was statistically assessed. Mycoplasma spp. was detected in 12.73% of samples. The frequency of the isolated bacteria was Escherichia coli (29.1%), Trueperella pyogenes (25.5%), Staphylococcus aureus (16.4%), Streptococcus agalactiae (9.1%), coagulase negative staphylococci (5.5%), and Corynebacterium pseudotuberculosis (1.8%) respectively. The significant difference between Saanen and Alpine was observed in antibiotic resistance to amoxicillin which showed more resistance in the Alpine breed. All the isolated bacteria showed multidrug resistance. Based on the obtained data using of antibiotics more accurate and using antibiogram test by clinician is necessary in the treatment of mastitis.

A Novel Polymerase Chain Reaction Assay for the Detection of Seven Mycoplasma Species of Cattle Origin

The aim of this study was to develop a pair of primers for detecting ruminant mycoplasma pathogens. We designed a set of primers based on the most similar sequences within 16sRNA regions of seven Mycoplasma spp. These primers have high sensitivity for detecting Mycoplasma dispar, M. arginine, M. canadense, M. bovis, M. alkalescens, M. californicum, and M. bovisgenitalium within the annealing temperature range of 46°C to 48°C. The minimum amount of DNA that can be detected using the protocol is 250 ng, which is equivalent to 2,000 cfu/mL. The primers can detect mycoplasma from DNA extracted directly from milk samples. The common bovine mastitis pathogens of Staphylococcus aureus coagulase-negative staphylococci, Escherichia coli, Streptococcus uberis, Klebiella pneumonia, Korucia rosea, and Acholeplasma spp. were not detected by the primers. We believe the high sensitivity and specificity of these primers make them useful for detecting infection with seven Mycoplasma species in ruminants, allowing the primers to be used in clinical settings.

Etiology of Bovine Mastitis

Mastitis in dairy animals is the primary concern of dairy farmers, which is the most common disease that causes huge economic losses in the dairy industry. The economic losses due to mastitis are from a reduction in milk yield, condemnation of milk with antibiotic residues, veterinary treatment costs, and death. In addition, some mastitis pathogens also cause serious human diseases associated with the contamination of milk or milk products with bacteria or their toxins. Bovine mastitis is mainly caused by a wide range of environmental and contagious bacterial mastitis pathogens. Contagious pathogens are those whose main reservoir is the infected udder. Contagious pathogens mainly spread among animals during milking process whereas environmental pathogens spread from environment to udder at any time. The source of the environmental pathogens is the surrounding environment of an animal. The major contagious pathogens include Staphylococcus aureus, Streptococcus agalactiae, and Mycoplasma spp. and the minor contagious pathogens include Corynebacterium bovis and others. Major environmental pathogens include coliform bacteria (Escherichia coli, Klebsiella spp., Enterobacter spp. and Citrobacter spp.), environmental streptococci (Strep. dysgalactiae, Strep. uberis). This chapter covers detailed review of published data on contagious and environmental pathogens responsible for bovine mastitis.

Antimicrobial Resistance Profiles and Genes in Streptococcus uberis Associated With Bovine Mastitis in Thailand

Streptococcus uberis is recognized as an environmental mastitis pathogen in dairy cattle. The varied success rate of antibiotic treatment for S. uberis intramammary infection may be associated with the antimicrobial resistance (AMR) of these bacteria. This observational study aimed to analyze 228 S. uberis strains associated with bovine mastitis in northern Thailand from 2010 to 2017. AMR and AMR genes were determined by the minimum inhibitory concentration (MIC) using a microdilution method and polymerase chain reaction, respectively. The majority of S. uberis strains were resistant to tetracycline (187/228, 82.02%), followed by ceftiofur (44/228, 19.30%), and erythromycin (19/228, 8.33%). The MIC50 and MIC90 of ceftiofur in 2017 were 2–4-fold higher than those in 2010 (P < 0.01). Resistance to tetracycline and ceftiofur significantly increased between 2010 and 2017 (P < 0.05). The most common gene detected in S. uberis was tetM (199/228, 87.28%), followed by ermB (151/228, 66.23 %) and blaZ (15/228, 6.58 %). The association between tetracycline resistance and tetM detection was statistically significant (P < 0.01). The detection rates of tetM significantly increased, while the detection rates of tetO and ermB significantly decreased during 2010–2017. AMR monitoring for bovine mastitis pathogens, especially S. uberis, is necessary to understand the trend of AMR among mastitis pathogens, which can help create an AMR stewardship program for dairy farms in Thailand.

Identification of bovine mastitis pathogens using MALDI-TOF mass spectrometry in Brazil

In this Research Communication we evaluate the use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to identify 380 bacteria isolated from cases of bovine mastitis in Brazil. MALDI-TOF MS identifications were compared to previous identifications by biochemical tests and 16S rRNA sequencing. MALDI-TOF MS achieved a typeability of 95.5%. The accuracy of MALDI-TOF MS for the identification of Staphylococcus isolates was 93.2%. The agreement between MALDI-TOF MS and biochemical identification of Streptococcus agalactiae was 96%, however, the agreement between these techniques for identifying other catalase-negative, Gram-positive cocci was lower. Agreement in identifying Gram-negative bacteria at the genus level was 90.5%. Our findings corroborate that MALDI-TOF MS is an accurate, rapid and simple technique for identifying bovine mastitis pathogens. The availability of this methodology in some research institutions would represent a significant step toward increasing the diagnosis and epidemiological studies of bovine mastitis and other animal infectious diseases in Brazil.

Silver and Copper Nanoparticles Inhibit Biofilm Formation by Mastitis Pathogens

Bovine mastitis is a common bovine disease, frequently affecting whole herds of cattle. It is often caused by resistant microbes that can create a biofilm structure. The rapidly developing scientific discipline known as nanobiotechnology may help treat this illness, thanks to the extraordinary properties of nanoparticles. The aim of the study was to investigate the inhibition of biofilms created by mastitis pathogens after treatment with silver and copper nanoparticles, both individually and in combination. We defined the physicochemical properties and minimal inhibitory concentration of the nanoparticles and observed their interaction with the cell membrane, as well as the extent of biofilm reduction. The results show that the silver–copper complex was the most active of all nanomaterials tested (biofilm was reduced by nearly 100% at a concentration of 200 ppm for each microorganism species tested). However, silver nanoparticles were also effective individually (biofilm was also reduced by nearly 100% at a concentration of 200 ppm, but at concentrations of 50 and 100 ppm, the extent of reduction was lower than for the complex). Nanoparticles can be used in new alternative therapies to treat bovine mastitis.

Genome-Wide Associations for Microscopic Differential Somatic Cell Count and Specific Mastitis Pathogens in Holstein Cows in Compost-Bedded Pack and Cubicle Farming Systems

The aim of the present study was to detect significant SNP (single-nucleotide polymorphism) effects and to annotate potential candidate genes for novel udder health traits in two different farming systems. We focused on specific mastitis pathogens and differential somatic cell fractions from 2198 udder quarters of 537 genotyped Holstein Friesian cows. The farming systems comprised compost-bedded pack and conventional cubicle barns. We developed a computer algorithm for genome-wide association studies allowing the estimation of main SNP effects plus consideration of SNPs by farming system interactions. With regard to the main effect, 35 significant SNPs were detected on 14 different chromosomes for the cell fractions and the pathogens. Six SNPs were significant for the interaction effect with the farming system for most of the udder health traits. We inferred two possible candidate genes based on significant SNP interactions. HEMK1 plays a role in the development of the immune system, depending on environmental stressors. CHL1 is regulated in relation to stress level and influences immune system mechanisms. The significant interactions indicate that gene activity can fluctuate depending on environmental stressors. Phenotypically, the prevalence of mastitis indicators differed between systems, with a notably lower prevalence of minor bacterial indicators in compost systems.

A Novel Polymerase Chain Reaction Assay for the Detection of Seven Mycoplasma Species of Cattle Origin

The aim of this study was to develop a pair of primers for detecting ruminant mycoplasma pathogens. We designed a set of primers based on the most similar sequences within 16sRNA regions of seven Mycoplasma spp. These primers have high sensitivity for detecting Mycoplasma dispar, M. arginine, M. canadense, M. bovis, M. alkalescens, M. californicum, and M. bovisgenitalium within the annealing temperature range of 46°C to 48°C. The minimum amount of DNA that can be detected using the protocol is 250 ng, which is equivalent to 2,000 cfu/mL. The primers can detect mycoplasma from DNA extracted directly from milk samples. The common bovine mastitis pathogens of Staphylococcus aureus coagulase-negative staphylococci, Escherichia coli, Streptococcus uberis, Klebiella pneumonia, Korucia rosea, and Acholeplasma spp. were not detected by the primers. We believe the high sensitivity and specificity of these primers make them useful for detecting infection with seven Mycoplasma species in ruminants, allowing the primers to be used in clinical settings.

Milk Lactose as a Biomarker of Subclinical Mastitis in Dairy Cows

Bovine subclinical mastitis can cause great harm to dairy herds because of its negative impact on milk production and quality and cow health. Improved diagnostic tools are needed to maximise the control of subclinical mastitis distribution and ensure the high quality of milk as an industrial product. Between 2015 and 2020, seventy-two dairy herds were screened for bovine subclinical mastitis causative agents to identify the relationship between seasons, lactose levels and subclinical mastitis infection. The predominant species found in the milk samples were mixed microbiota, coagulase-negative Staphylococcus and Staphylococcus aureus. Yeasts were found exclusively in autumn, while Enterococcus spp. and Escherichia coli were only found in summer and autumn. A negative correlation was detected between milk lactose and number of somatic cells in milk (−0.471; p < 0.001). The lactose levels in milk were closely associated with the prevalence (%) of subclinical mastitis pathogens, such as Streptococcus agalactiae (y = −1.8011x + 10.867, R2 = 0.9298), Staph. aureus (y = −3.5216x + 25.957, R2 = 0.8604) and other Streptococci (y = −0.5956x + 7.6179, R2 = 0.6656). These findings suggest that milk lactose may be used as a biomarker of suspected udder inflammation in modern health prevention programmes for cows to reduce the prevalence of subclinical mastitis pathogens in dairy cattle herds.