Clinical Mastitis Cows Research Articles

Evaluation of Lactic Acid Bacteria and Total Bacterial Load in Milk from Clinical Mastitis, Subclinical Mastitis and Healthy Cows

Aim: Studies on alternative preventive and therapeutic approaches for bovine mastitis have gained momentum in the veterinary field due to bacterial antibiotic resistance. In this study; It was aimed to determine the total bacterial counts and lactic acid bacteria counts in milk taken from lactating cows, clinical/subclinical mastitis and healthy udder lobes, compare them with the results of the California Mastitis Test, used a screening test, and determine the variability of lactic flora load in the cases. Materials and Methods: The milk samples were collected from lactating cows from fifteen farms. Udder quarters were categorized into three subgroups: non-mastitis (NM; n=35), clinical mastitis (CM; n=30), and subclinical mastitis (SCM; n=31). Total bacteria and lactic acid bacteria were counted by inoculating milk samples onto MRS (De Man Rogosa Sharpe Agar), M17 agar and PCA (Plate Count Agar) media. Results: No significant difference between the three groups regarding the logarithmic averages of MRS and M17 colony numbers were found. When evaluated using MRS and M17, no significant difference existed among the NM, SCM, and CM groups (P=0.093). Positive correlations (P=0.001) existed between the mean of PCA, CMT, MRS and M17 media. Conclusion: The total bacterial count in milk, as well as the LAB load, are affected by the growing environment and environmental conditions. The milk microbiota is altered in healthy udder quarters and clinical and subclinical mastitis cases. Further investigation is needed to understand the flora characteristics of milk in cases of bovine mastitis and healthy animals.

Inflammatory Signaling via PEIZO1 Engages and Enhances the LPS-Mediated Apoptosis during Clinical Mastitis.

Bovine clinical mastitis is characterized by inflammation and immune responses, with apoptosis of mammary epithelial cells as a cellular reaction to infection. PIEZO1, identified as a mechanotransduction effector channel in nonruminant animals and sensitive to both mechanical stimuli or inflammatory signals like lipopolysaccharide (LPS). However, its role in inflammatory processes in cattle has not been well-documented. The aim of this study was to elucidate the in situ expression of PIEZO1 in bovine mammary gland and its potential involvement in clinical mastitis. We observed widespread distribution and upregulation of PIEZO1 in mammary epithelial cells in clinical mastitis cows and LPS-induced mouse models, indicating a conserved role across species. In vitro studies using mammary epithelial cells (MAC-T) revealed that LPS upregulates PIEZO1. Notably, the effects of PIEZO1 artificial activator Yoda1 increased apoptosis and NLRP3 expression, effects mitigated by PIEZO1 silencing or NLRP3 inhibition. In conclusion, the activation of the PIEZO1-NLRP3 pathway induces abnormal apoptosis in mammary epithelial cells, potentially serving as a regulatory mechanism to combat inflammatory responses to abnormal stimuli.

Early Detection and Prevention of Sub - Clinical Mastitis in Cows Using TANUVAS - Masti-Guard Kit

Early Detection and Prevention of Sub - Clinical Mastitis in Cows Using TANUVAS - Masti-Guard Kit

Virulence and antimicrobial resistance genes detected in Staphylococcus spp. isolated from clinical and non-clinical mastitis using whole-genome sequencing

Staphylococcus spp. are among the most isolated bacteria in clinical and subclinical mastitis cases in dairy cattle. The genus comprises biofilm-forming bacteria capable of producing toxins and acquiring resistance to multiple drugs. This work aimed to evaluate the genetic profile related to virulence and antimicrobial resistance characteristics of Staphylococcus spp., isolated from clinical mastitis and non-clinical fresh cows using whole genome sequencing (WGS). The bacterial collection consisted of 29 Staphylococcus strains isolated from clinical cases of mastitis (n = 7), as well as milk samples collected from fresh cows (n = 22). Strains were identified as Staphylococcus aureus (n = 2), Staphylococcus chromogenes (n = 19), and Staphylococcus haemolyticus (n = 8). A total of 94 virulence genes were observed, including pvl, icaA, icaD genes, and microbial surface components that recognize adhesive matrix molecules (MSCRAMMs). We also detected important resistance genes such as blaZ, ant(4), erm(B), fexA, lnu(D), tet(L), and tet(M). The phylogenetic tree listed the species as expected and presented four clades. A variety of virulence and resistance genes were detected. In addition, the expression of important genes such as those responsible for the formation of biofilms and enterotoxins may represent a risk to the health of consumers. being a concern for public health.

Effect of Photobiomodulation With Laser on The Composition, Production and Quality of The Milk of Cows with Clinical Mastitis

Objective: This study assesses the impact of LASER photobiomodulation (808nm at 1400 mW and 637nm at 100 mW, with a 2-minutes irradiation per wavelength, total dose: 1.4 W/cm² and 1 W/cm², respectively) on milk composition and quality in cows suffering from clinical mastitis. Theoretical Framework: Research indicates LASER photobiomodulation's potential in improving animal health, yet there’s a gap in specific therapeutic protocols for treating clinical mastitis in cows using this technology. Methodology: The study involved 20 cows with clinical mastitis. Their mammary quarters were assessed for fat and protein composition, Somatic Cell Count, Urea Nitrogen levels, Microbiological Culture, and Antibiotic Sensitivity and Milk production. Comparisons were made between two groups over six days: one receiving a combination of antibiotics and LASER photobiomodulation therapy, and a control group receiving only antibiotics. Results and Conclusion: LASER photobiomodulation significantly aided the treatment of clinical mastitis in conjunction with antibiotics. This combination not only helped in curing mastitis but also reduced the Somatic Cell Count and increased milk production, marking it as a viable alternative therapy. Implications of the Research: This study contributes to establishing a more effective treatment protocol for clinical mastitis, combining photobiomodulation with traditional antibiotic methods. Originality/Value: This pioneering research showcases the use of a photobiostimulation device as a supplementary treatment for controlling clinical mastitis in dairy cows, integrated with antibiotic therapy. It underscores the enhanced efficiency of such combined treatments, contributing to the Sustainability.

Clinical and Subclinical Bovine Mastitis: Staphylococcus aureus Isolation and Identification from Dairy Farms Located in and Around Hawassa Town, Southern Ethiopia

BACKGROUND: Mastitis is an expensive disease of dairy cattle in most countries, including Ethiopia. The most commonly recovered bacterial pathogen from mastitis milk is _Staphylococcus aureus_ in dairy herds worldwide, with huge economic losses in the dairy industry. OBJECTIVE: To isolate and identify _Staphylococcus aureus_ from bovine mastitis milk. Additionally, risk factors associated with the occurrence of mastitis were determined. METHODS: A cross-sectional study was conducted from March 2021 to August 2021 on dairy farms in and around Hawassa town. A total of 250 lactating cows were examined for clinical and subclinical mastitis from randomly selected dairy farms. Clinical signs and the California Mastitis Test (CMT) were used to identify clinical and subclinical mastitis, respectively. RESULTS: During the study period, 50.8% of cows had mastitis, of which 4.8% and 46% showed clinical and subclinical mastitis, respectively. The quarter-level prevalence was 27.4%; of which the clinical form was 2.9%, while the subclinical mastitis was 24.5%. Logistic regression analysis showed a significant association among cows of different age groups, lactation stages, and farm hygienic status with the occurrence of mastitis (p < 0.05). A bacteriological study targeting _S. aureus_ was conducted with all (n=127) milk samples collected from clinical and subclinical mastitis cows. Bacterial identification targeting _S. aureus_ was done, and this agent was identified in 60 (47.2%) milk samples. This pathogen was found to be higher (47.8%) in subclinical than in clinical (41.6%) mastitis. CONCLUSION: This study showed that mastitis was prevalent in dairy cattle of the study area, with a higher case of _S. aureus_ in subclinical mastitis. However, the recovery of _S. aureus_ in nearly half of the mastitis milk indicated the possible presence of other pathogens. Therefore, further study to recover other potential pathogens commonly causing mastitis can be a good approach.

To study the sensitivity of field strains of <i>Str. agalactiae</i> and <i>Str. uberis </i>— causative agents of cow mastitis to a drug based on marbofloxacin

An increase in the number of resistant strains of mastitis pathogens Str. agalactiae and Str. uberis is one of the causes of massive subclinical and clinical mastitis in cows. The aim of the study was to study the sensitivity of field strains of Str. Agalactiae and Str. uberis to the drug based on marbofloxacin «Marboflocin 10%» (LLC «AleksAnn»), as well as to a number of the most commonly used antibacterial drugs (ABP). Sensitivity to marbofloxacin was determined by the method of serial dilutions, to other antibacterial drugs — by the disco-diffusion method. 15 samples of breast secretions from cows with a clinical form of mastitis were examined, from which 15 cultures of streptococci were isolated, including Str. agalactiae and Str. uberis — in 53.3% of the samples. All isolated cultures of Str. Agalactiae and Str. uberis were susceptible to marbofloxacin. The mean value of the minimum inhibitory concentration (MIC) of marbofloxacin for cultures of Str. uberis is 0.50 µg/ml, and for Str. agalactiae — 0.44 ± 0.13 µg/ml. To other antibacterial drugs, the studied cultures of Str. uberis and Str. agalactiae showed different sensitivity, resistance to amoxicillin was revealed, incl. with clavulanic acid, tylosin

Bovine mastitis: Prevalence, causes and associated risk factors in Silte Zone, Ethiopia

Mastitis is an important disease of dairy cows, and it causes huge economic losses to dairy farm owners due to a decrease in milk production, and reduction in milk quality, and an increase in the cost of cow treatment. A crosssectional study was conducted in Southern Ethiopia’s Silte zone from October 2020 to June 2021 aimed to estimate mastitis prevalence, assess related risk factors, and identify prevalent bacterial causes. Three hundred eighty-four lactating cows were examined for abnormalities in udder quarters and teats. Milk samples were tested for subclinical mastitis via the California mastitis test (CMT) and cultured for causative agents from clinical mastitic and CMT-positive cows. The overall prevalence of mastitis was 54.9% (95% CI=49.9-59.9), of which 52.1% (95% CI=47.1-57.1) was subclinical and 2.9% (95% CI=1.6 -5.1) clinical mastitis. From a total of 1536 quarters examined 41 (2.7%) quarters were found blind and 427 (27.7%) quarters were affected by mastitis. Overall, more hind quarters, 236 (55.5%) were affected than the front quarters, 189 (44.5%) of udder. The prevalence of mastitis was significantly higher during early lactation, ≤4 months (p< 0.05); and it was increased with increasing parity (p< 0.05) and age (p< 0.05). Cows with no bedding were more affected than those with bedding (p< 0.05). Multivariable logistic regression showed that cows with round and flat teat ends were 2.84 and 11.85 times more likely to contract mastitis. Also, cows with pendulous udder, producing more than 10 liters per day milk and during the early 4 months of lactation were 1.87, 6.81, and 2.14 more likely affected by mastitis than normal udder, producing less than 10 liters of milk per day and lactation after five months, respectively. Milk samples collected from 211 mastitis-positive cows were cultured using standard bacteriological technique, and the isolated bacteria were Staphylococcus aureus (29.5%), Staphylococcus epidermidis (14.2%), Staphylococcus intermedius (11.6%), Staphylococcus hyicus (11.1%), Streptococcus agalactiae (8.9%), Streptococcus disgalactiae (6.3%), E. coli (5.8%), Streptococcus uberis(5.3%), Klebsiella spp. (1.6%) and Enterococcus spp. (1.1%). Owners should givepro per udder care and are advised to apply dry cow therapy. Extension workers should raise awareness.

Comparative analysis of total protein, casein, lactose, and fat content in milk of cows suffering from subclinical and clinical mastitis caused by Streptococcus spp.

The aim of the study was to analyse the total protein (TP), casein (CAS), lactose (LAC), and fat content of milk from cows with subclinical (SCM) and clinical mastitis (CM) caused by Streptococcus spp. A total of 60 milk samples from diseased cows and 30 milk samples from healthy cows were included in the study. Milk samples were taken from Holstein-Friesian cows from four dairy farms in Lublin Province. The bacteriological examination of the milk was performed and the somatic cells count in 1 mL of milk was determined using a SomaCount FC automatic cell counter. Determination of TP, CAS, LAC, FAT and FA levels in milk was carried out using a DairySpec FT automated Fourier transform infrared spectrometer. Total protein in milk from HE was significantly higher than in milk from cows with mastitis (4.04% vs 3.57% in milk from SCM cows and 3.7% in milk from CM cows, P = 0.001). The CAS level was 2.73% in milk from CM cows and 2.92% in milk from SCM cows vs 3.30% in milk from HE cows, P = 0.001. The changes in CAS and TP in milk resulted in a significant difference in the CAS/TP ratio (81.7% in milk from HE cows vs 73.8% in milk from CM cows). A decrease in levels was also recorded for LAC (4.8% in milk from HE cows vs 4.51% in milk from SCM cows and 4.01% in milk from CM cows, P = 0.001). The fat level was significantly higher in milk from healthy cows than in milk from cows with mastitis (4.0% vs 2.3% in milk from SCM cows and 1.64% in milk from CM cows, P = 0.001). It should be emphasised that the decrease in the levels of TP, LAC and FAT was significant not only in milk from CM cows but also in milk from SCM cows. This is very unfavourable, because the reduction in the main milk components results in poor quality dairy products and impairs line processes.

Clinical Mastits Incidence In Small-Scale Dairy Cow Farms

Abstract The incidence of clinical mastitis (CM) in small and large-scale dairy cow farms can be highly variable and can be affected by age, parity, post-calving status, and atmospheric conditions. The current study aimed to assess the CM-incidence and its association with the post-calving days, number of lactations, season, and number of affected udder quarters in dairy cows in small-scale dairy farms. The study was conducted within one calendar year in 177 small-scale farms with 864 dairy cows. Clinically confirmed CM cows (n=72) were sampled from each udder quarter and processed for bacteriology examination. The CM-positive samples were grouped according to the season (spring, summer, autumn, and winter), the number of days after calving (<90, 90-180, and >180), the number of lactations (1-st, 2-5-th, and >6-th), and the number of affected udder quarters (one, two, three, and four). The CM-positive samples (n=72, 8.3%) were confirmed on bacteriology examination in significantly lower count (n=56, 6.5%). The 2-5-th lactations cows (n=35, 68.6%) were significantly more compared to the first lactation (n=6, 11.8%), and >6-th lactation cows (n=10, 19.6%). CM cows with infection of one udder quarter (n=40, 78.4%) were significantly more than the cows with two (n=6, 11.8%), and four infected quarters (n=5, 9.8%). The CM-incidence in small-scale dairy cow farms in North Macedonia was 8.3% and 6.5% by clinical and bacteriology examination, respectively. The highest CM susceptibility was observed in the cows between the second and fifth lactations. One udder quarter was most frequently affected in CM-positive cases.

Comparative Proteomic Analysis of Milk-Derived Extracellular Vesicles from Dairy Cows with Clinical and Subclinical Mastitis.

Extracellular vesicles (EVs) are membranous vesicles found in biological fluids with essential functions. However, milk-derived EV proteins from clinical mastitis (CM) and subclinical mastitis (SM) cows have yet to be studied in detail. In this study, milk-derived EVs of CM, SM, and Healthy cows were extracted using a combination of acetic acid/ultracentrifugation and density gradient ultracentrifugation and analyzed using a shotgun proteomic by data-independent acquisition mode. A total of 1253 milk exosome proteins were identified and quantified. Differently enriched (DE) proteins were identified as given a Benjamini−Hochberg adjusted p < 0.05 and a fold change of at least 2. There were 53 and 1 DE proteins in milk-derived EVs from CM and SM cows compared with healthy cows. Protein S100-A9, Protein S100-A8, Chitinase-3-like protein 1, Haptoglobin, Integrin beta-2, and Chloride intracellular channel protein 1 were more abundant in the CM group (adjusted p < 0.05). Still, their enrichment in the SM group was not significant as in the Healthy group. The enrichment of DE proteins between CM and Healthy group was consistent with elevated GO (Gene Ontology) processes—defense response, defense response to Gram-positive bacterium, granulocyte chemotaxis also contributed to Reactome pathways—neutrophil degranulation, innate immune system, and antimicrobial peptides in the CM group. These results provide essential information on mastitis-associated proteins in milk-derived EVs and indicate the biological functions of milk-derived EVs proteins require further elucidation.

Effect of low frequency oscillations during milking on udder temperature and welfare of dairy cows

The study aimed to investigate the effect of low-frequency oscillations on the cow udder, milk parameters, and animal welfare during the automated milking process. The study's objective was to investigate the impact of low-frequency oscillations on the udder and teats' blood circulation by creating a mathematical model of mammary glands, using milkers and vibrators to analyze the theoretical dynamics of oscillations. The mechanical vibration device developed and tested in the study was mounted on a DeLaval automatic milking machine, which excited the udder with low-frequency oscillations, allowing the analysis of input parameters (temperature, oscillation amplitude) and using feedback data, changing the device parameters such as vibration frequency and duration. The experimental study was performed using an artificial cow's udder model with and without milk and a DeLaval milking machine, exciting the model with low-frequency harmonic oscillations (frequency range 15-60 Hz, vibration amplitude 2-5 mm). The investigation in vitro applying low-frequency of the vibration system's first-order frequencies in lateral (X) direction showed the low-frequency values of 23.5-26.5 Hz (effective frequency of the simulation analysis was 25.0 Hz). The tested values of the first-order frequency of the vibration system in the vertical (Y) direction were 37.5-41.5 Hz (effective frequency of the simulation analysis was 41.0 Hz), with higher amplitude and lower vibration damping. During in vivo experiments, while milking, the vibrator was inducing mechanical milking-similar vibrations in the udder. The vibrations were spreading to the entire udder and caused physiotherapeutic effects such as activated physiological processes and increased udder base temperature by 0.57°C (p < 0.001), thus increasing blood flow in the udder. Used low-frequency vibrations did not significantly affect milk yield, milk composition, milk quality indicators, and animal welfare. The investigation results showed that applying low-frequency vibration on a cow udder during automatic milking is a non-invasive, efficient method to stimulate blood circulation in the udder and improve teat and udder health without changing milk quality and production. Further studies will be carried out in the following research phase on clinical and subclinical mastitis cows.

Effect of anemoside B4 on milk whey in clinical mastitis-affected cows elucidated using tandem mass tag (TMT)-based quantitative proteomics

Intramuscular injection of anemoside B4 (AB4) has a superior therapeutic effect on clinical mastitis in lactating cows. Here, we explored AB4’s effect on milk whey in clinical mastitis-affected cows using proteomics. Among fifty clinical mastitis cows received AB4 administration (0.05 ml/kg/day, for 7 days), twelve healed cows were selected and marked as group T. Twelve clinically heathy cows received the same dose of saline for 7 days, marked as group C. Collected milk whey of group T before and after AB4 administration marked as T1 and T2, respectively. The milk whey of group C after saline injection marked as C1. Milk whey protein changes were detected using tandem mass tag-based quantitative proteomic. We identified 872 quantifiable proteins in the samples. Among them, 511 proteins between T1 and C1, and 361 proteins between T2 and T1 were significantly altered. T1 than C1 had significantly more proteins associated with inflammatory damage and trans-endothelial migration of leukocytes, whereas these proteins were reduced in T2 treated with AB4. Compared with C, proteins associated with fibrin clot degradation and complement system activation were downregulated in T1 but upregulated in T2. In summary, AB4 can exert its therapeutic effect on clinical mastitis in cows mainly by reducing inflammatory damage, activating the complement system, inhibiting trans-endothelial migration of leukocytes, and promoting degradation of milk fibrin clots.

Genotypic and Phenotypic Characterization of Mammaliicoccus sciuri - A MDR Strain causing Clinical Mastitis in Cows of Odisha, India

Background: Mammaliicoccus sciuri is a multidrug resistant human pathogen. Literature on its role causing bovine mastitis is scarce. Detection of M. sciuri from a cow with clinical mastitis in Malkangiri district of Odisha stimulated to ascertain its status in a larger population and to study its characteristics in genotypic and phenotypic levels. Methods: 520 lactating cows from various herds of Malkangiri district of Odisha, India were screened for presence of clinical mastitis. Milk samples were collected aseptically from 15 cows with clinical mastitis. 16S rRNA amplification was performed to know presence of bacterial pathogen (s). Conventional PCR was carried out to detect resistance genes against β-lactam (blaZ), aminoglycoside (aacA-aphD) and tetracycline (tetK, tetM). In vitro sensitivity test was studied against M. sciuri isolates using nine antibiotic discs. Phylogenetic analysis was performed to compare its origin. Result: M. sciuri was isolated in 6 (40.0%) mastitis positive milk samples. In vitro antibiotic sensitivity test against M. sciuri isolates showed highest degree of resistant to penicillin (100%) followed by, cefoperazone (33.3%), streptomycin (33.3%), tetracycline (16.0%). One isolate of M. sciuri had tetK, tetM resistance gene. Phylogenetic analysis showed (75-100%) similarity with other isolates obtained from GeneBank. Present study highlights tetK gene in M. sciuri.

Discrepancies among healthy, subclinical mastitic, and clinical mastitic cows in fecal microbiome and metabolome and serum metabolome.

Mastitis is generally considered a local inflammatory disease caused by the invasion of exogenous pathogens and resulting in the dysbiosis of microbiota and metabolites in milk. However, the entero-mammary pathway theory may establish a possible link between some endogenous gut bacteria and the occurrence and development of mastitis. In the current study, we attempted to investigate differences in the gut microbiota profile and metabolite composition in gut and serum from healthy cows and those with subclinical mastitis and clinical mastitis. Compared with those of healthy cows, the microbial community diversities in the feces of cows with subclinical mastitis (SM) and clinical mastitis (CM) were lower. Lower abundance of Bifidobacterium, Romboutsia, Lachnospiraceae_NK3A20_group, Coprococcus, Prevotellaceae_UCG-003, Ruminococcus, and Alistipes, and higher abundance of the phylum Proteobacteria and the genera Escherichia-Shigella and Streptococcus were observed in CM cows. Klebsiella and Paeniclostridium were significantly enriched in the feces of SM cows. Several similarities were observed in feces and serum metabolites in mastitic cows. Higher levels of proinflammatory lipid products (20-trihydroxy-leukotriene-B4, 13,14-dihydro-15-keto-PGE2, and 9,10-dihydroxylinoleic acids) and lower levels of metabolites involved in secondary bile acids (deoxycholic acid, 12-ketolithocholic acid), energy (citric acid and 3-hydroxyisovalerylcarnitine), and purine metabolism (uric acid and inosine) were identified in both SM and CM cows. In addition, elevated concentrations of IL-1β, IL-6, tumor necrosis factor-α and decreased concentrations of glutathione peroxidase and superoxide dismutase were detected in the serum of SM and CM cows. Higher serum concentrations of triglyceride and total cholesterol and lower concentrations of high-density lipoproteins in mastitic cows might be related to changes in the gut microbiota and metabolites. These findings suggested a significant difference in the profile of feces microbiota and metabolites in cows with different udder health status, which might increase our understanding of bovine mastitis.

Clinical mastitis in cows and their response to Invitro sensitivity

Clinical mastitis in cows and their response to Invitro sensitivity

Clinical mastitis in cows and their response to Invitro sensitivity

A total of 50 cases of bovine clinical mastitis in Tripoli were subjected to microbiological examination. Thirty-five bacterial isolates were obtained and further identified by using the biochemical tests. Staphylococcus spp. (36%) was the predominant causative organism, then E. coli (24%) and Streptococcus spp. (6%) Additionally, twelve cases were caused by nonbacterial agents. The bacteria isolates were tested for their in-vitro susceptibility to different antimicrobial agents that are used in commercial intramammary infusion products. Antibacterial susceptibility testing showed that the bacteria isolates were sensitive to ciprofloxacin (100%), enrofloxacin (96%), cefotaxime (90%), deoxicillin (88.8%), clorophenicol (66.5), ampicillin (62.5%), amoxicillin (50%), vancomycin (42%) and fusidic acid (33.3%). According to these results, the ciprofloxacin was proved to be the drug of choice.

Induction of mastitis by cow-to-mouse fecal and milk microbiota transplantation causes microbiome dysbiosis and genomic functional perturbation in mice

BackgroundMastitis pathogenesis involves a wide range of opportunistic and apparently resident microorganims including bacteria, viruses and archaea. In dairy animals, microbes reside in the host, interact with environment and evade the host immune system, providing a potential for host-tropism to favor mastitis pathogenesis. To understand the host-tropism phenomena of bovine-tropic mastitis microbiomes, we developed a cow-to-mouse mastitis model.MethodsA cow-to-mouse mastitis model was established by fecal microbiota transplantation (FMT) and milk microbiota transplantation (MMT) to pregnant mice to assess microbiome dysbiosis and genomic functional perturbations through shotgun whole metagenome sequencing (WMS) along with histopathological changes in mice mammary gland and colon tissues.ResultsThe cow-to-mouse FMT and MMT from clinical mastitis (CM) cows induced mastitis syndromes in mice as evidenced by histopathological changes in mammary gland and colon tissues. The WMS of 24 samples including six milk (CM = 3, healthy; H = 3), six fecal (CM = 4, H = 2) samples from cows, and six fecal (CM = 4, H = 2) and six mammary tissue (CM = 3, H = 3) samples from mice generating 517.14 million reads (average: 21.55 million reads/sample) mapped to 2191 bacterial, 94 viral and 54 archaeal genomes. The Kruskal–Wallis test revealed significant differences (p = 0.009) in diversity, composition, and relative abundances in microbiomes between CM- and H-metagenomes. These differences in microbiome composition were mostly represented by Pseudomonas aeruginosa, Lactobacillus crispatus, Klebsiella oxytoca, Enterococcus faecalis, Pantoea dispersa in CM-cows (feces and milk), and Muribaculum spp., Duncaniella spp., Muribaculum intestinale, Bifidobacterium animalis, Escherichia coli, Staphylococcus aureus, Massilia oculi, Ralstonia pickettii in CM-mice (feces and mammary tissues). Different species of Clostridia, Bacteroida, Actinobacteria, Flavobacteriia and Betaproteobacteria had a strong co-occurrence and positive correlation as the indicator species of murine mastitis. However, both CM cows and mice shared few mastitis-associated microbial taxa (1.14%) and functional pathways regardless of conservation of mastitis syndromes, indicating the higher discrepancy in mastitis-associated microbiomes among lactating mammals.ConclusionsWe successfully induced mastitis by FMT and MMT that resulted in microbiome dysbiosis and genomic functional perturbations in mice. This study induced mastitis in a mouse model through FMT and MMT, which might be useful for further studies- focused on pathogen(s) involved in mastitis, their cross-talk among themselves and the host.Graphical

Prevalence and Phylodiversity of ESBL-Producing Coliforms Isolated from Ruminant Mastitis in Nigeria

Abstract The public health threat posed by Extended-spectrum beta-lactamase producing E. coli (ESBL-EC) in food animal production systems has attracted global attention. Data on the prevalence, diversity and genetic characteristics of ESBL-producing coliforms are key to advocacy on promoting responsible antimicrobial stewardship and proper planning of control strategies. The coliforms were isolated from 1052 milk samples of 160 cows, 103 ewes and 103 does with mastitis in Plateau State, Nigeria and analysed for ESBL production by phenotypic, biochemical, antimicrobial sensitivity and genetic characterization. The percentage of occurrence of clinical mastitis in cows, ewes, and does were 0.2 %, 0 %, 1.5 % respectively, while the percentage occur-rence of subclinical mastitis in ruminants were 18.1 %, 28.2 % and 38.3 % respectively. From the 677 isolates, 31.3 % (n = 212) were ESBL producing coliforms, with a prevalence of 48.6 %, 18.4 %, 12.7 %, 8.9 %, 5.7 %, 3.8 % and 1.9 % for E. coli, K. pneumoniae, C. freundii, K. aerogenes, S. marcescens, K. oxytoca and E. cloacae, respectively. The genetic characterization revealed a higher prevalence of bla CTX-M than bla TEM in the samples analysed (24.39 % vs. 12.19 %). High pairwise identity was observed among the bla CTX-M and bla TEM gene sequences obtained in this study, but they displayed high phylodiversity with sequences from ruminants and humans from other climes. The bla SHV gene was not detected. Multidrug resistances especially to the commonly used antimicrobials; ofloxacin, gentamycin and streptomycin in veterinary practice in Nigeria were observed. This has public health implications considering the fact that consumption of raw unpasteurized milk is a common practice in some cultures in Nigeria. Such practise will facilitate the transfer of multidrug resistant coliforms to humans resulting in the complications of treatment outcomes. To the best of our knowledge this is the first genetic characterization of ESBL-producing agents from ruminant mastitis in Nigeria.

Haptoglobin and Amyloid A Levels in Milk of Clinical and Subclinical Mastitic Cows in Turkey

Haptoglobin and Amyloid A Levels in Milk of Clinical and Subclinical Mastitic Cows in Turkey