California Mastitis Test Research Articles

Molecular insights into antimicrobial resistant Staphylococcus aureus strains: A potential zoonosis of goat origin

Antimicrobial-resistant (AMR) Staphylococcus aureus (S. aureus) strains have attained global attention due to their life-threatening zoonotic nature. Being a member of ESKAPE group, S. aureus has an ability to escape the biocidal action of antimicrobial drugs. The current study investigated the prevalence and molecular characterization of methicillin-resistant S. aureus (MRSA), β-lactam-resistant S. aureus (BRSA), aminoglycoside-resistant S. aureus (ARSA), tetracycline-resistant S. aureus (TRSA), and fluoroquinolones-resistant S. aureus (FRSA) associated with goat subclinical mastitis (SCM). Furthermore, the antimicrobial resistance and susceptibility profile of various antibiotics and non-antibiotics (NSAIDs, nisin, N-acetylcysteine, vitamin-C) along with their possible role in modulating the antibiotic resistance of MDR isolates was also investigated. A total of 768 goat milk samples were subjected to California mastitis test for SCM followed by bacteriological and molecular characterization of S. aureus. Moreover, in-vitro susceptibility of resistant antibiotics, non-antibiotics, and their combination against MDR S. aureus were conducted through well diffusion and broth microdilution assays. The results depicted that 55.47% and 26.82% of milk samples were positive for SCM and S. aureus, respectively. The molecular assay confirmed 35.92% of isolates as MRSA, 45.63% as BRSA, 50.49% as ARSA, and 32.52% but no isolate was confirmed as FRSA on molecular basis. The multidrug resistance was observed in 62.13% and 47.09% isolates, respectively. Molecular characterized MDR S. aureus revealed high homology of study isolates with the isolates of neighboring countries like India, Korea, Iran, and China. Antimicrobial susceptibility trials on well diffusion assay showed higher efficacy of different non-antibiotics with resistant antibiotics as penicillin with ketoprofen and gentamicin with flunixin meglumine while oxytetracycline with N-acetylcystiene. The synergy testing by checkerboard assay revealed synergistic activity of penicillin with ketoprofen, gentamicin with flunixin meglumine, and oxytetracycline with N-acetylcysteine. The current study highlighted the emergence and spread of AMR S. aureus strains from goat SCM and provided insights into possible drug repurposing of various non-antibiotics to modulate the multidrug resistance of S. aureus which will be helpful in devising the therapeutic options and control measures for this pathogen.

Prevalence, Risk Factors, and Antimicrobial Resistance of Staphylococcus and Streptococcus Species Isolated from Subclinical Bovine Mastitis.

Subclinical mastitis (SCM) is a prevalent serious disease among dairy cows worldwide. It poses a significant challenge to the dairy industry, animal welfare, and a threat to public health. The present study aimed to investigate the molecular detection, prevalence, and antimicrobial resistance of Staphylococcus spp. and Streptococcus spp. isolated from raw composite milk samples obtained from SCM dairy cattle in Bangladesh. A total of 612 quarters milk samples obtained from 153 cows were analyzed for SCM using the California Mastitis Test. Bacterial isolation and identification were carried out and bacterial species were confirmed using molecular polymerase chain reaction methods. Antimicrobial susceptibility testing was performed using disc diffusion method. The findings revealed that the prevalence of SCM was 70.3% (26/37), 35.95% (55/153), and 23.04% (141/612) in the herd, cow, and quarter levels, respectively. Among the positive samples, 92.7% (51/55) were Staphylococcus spp. (S. aureus, S. chromogenes, and S. simulans) and the remaining isolates were 7.3% (4/55) Streptococcus spp. (Streptococcus agalactiae and Streptococcus dysgalactiae). The most prevalent species was S. chromogenes, accounting for 67.3% (37/55). Antimicrobial susceptibility testing showed that 65.5% of isolates were susceptible to cefoxitin, whereas, 89.1% were resistant to penicillin. Overall, 12 isolates (21.8%) out of 55 were resistant to more than three classes of antimicrobials and were defined as multidrug-resistant isolates. Methicillin-resistance gene was detected in 61.1% of the cefoxitin-resistant isolates. A multivariate logistic regression analysis identified five potential risk factors including the lack of post-milking teat disinfection (OR: 3.06), absence of immediate feeding after milking (OR: 9.81), poor udder hygiene (OR: 7.83), tick infestation (OR: 13.76), and absence of dry cow therapy (OR: 3.31). The findings of the current study underscore the urgent requirement for targeted interventions, considering the identified factors to effectively manage and control SCM in dairy cows.

Comparative Evaluation of Antibiotic Therapies for Subclinical Staphylococcal Mastitis in Goats

Subclinical mastitis (SCM) is characterized by inflammation of the udder without visible signs of inflammation in the animal, udder and milk and hence could be diagnosed by laboratory examination only. Effective treatment of SCM in goats is crucial for maintaining quality milk production. This study was conducted at the Department of Veterinary Medicine, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University, Jabalpur (M.P.), India. A total of 236 lactating goats irrespective of age, breed, parity and stage of lactation, from both organized and unorganized sectors in and around Jabalpur, were screened over a period of six month. Based on modified California mastitis test scores, 83 goats tested positive for subclinical mastitis resulting in an overall occurrence of 35.17% (83/236). A total of 48 milk samples that tested positive for modified California mastitis test were further analyzed for bacterial isolation and identification. Among them, 34 (70.83%) samples were positive for only Staphylococcus spp., while 12 (25%) samples exhibited mixed infection with Staphylococcus spp. and Gram-positive bacilli. Then, 18 goats, positive for subclinical Staphylococcal mastitis were divided into three groups i.e. G1, G2 and G3 with six goats in each group for therapeutic study and six apparently healthy goats were taken as healthy control group (G4). G1 Goats were treated with Inj. Enrofloxacin LA @ 7.5 mg/kg b. wt. I/M single dose; G2 with Inj. Ceftizoxime @ 10 mg/kg b. wt. I/V single dose and G3 with Inj. Oxytetracycline LA @ 20 mg/kg b. wt. I/M single dose. The responses of therapeutic assessment were evaluated on the basis of somatic cell count and modified California mastitis test score. The most effective response for subclinical Staphylococcal mastitis in goats was observed with Ceftizoxime followed by Enrofloxacin LA and the least with Oxytetracycline LA.

Therapeutic Efficacy of Eucalyptus Oil as Teat Dip in Subclinical Mastitis in Goats

Mastitis, an inflammation of the parenchyma of mammary gland, results in alterations to the chemical and physical composition of milk due to a variety of infectious and non-infectious causes. The present work was conducted in the Department of Veterinary Medicine, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University, Jabalpur, Madhya Pradesh (M.P.). A total of 256 lactating goats irrespective of age, breed, parity and stage of lactation belonging to both organized and unorganized sectors at Jabalpur were screened from “February 2024 to July 2024”. On the basis of modified California mastitis test (MCMT) score, out of 256 lactating goats, 98 goats were found positive for subclinical mastitis with overall occurrence as 38.28% on animal basis and 33.33% on udder halves basis. Therapeutic study on 24 milk samples positive for subclinical mastitis by MCMT was conducted and categorized into four treatment groups (G1, G2, G3 and G4), each group comprised of 6 goats. Six apparently healthy goats were selected as healthy control group (G5). Two different teat dips viz. Eucalyptus oil 98% + DMSO 2% BID for 14 days in groups G1 and G3 and 5% Povidone iodine BID for 14 days in groups G2 and G4 were used as teat dips, respectively. Moreover, goats of groups G3 and G4 were commonly treated with parenteral antibiotic amoxicillin sulbactam @10 mg/kg b.wt. I/M OD for 3 days. Significantly higher values of somatic cell count, standard plate count, pH, freezing point and significantly lower value of lactose were observed in subclinical mastitis affected goats. The mean density and mean water percentage values varied within normal physiological range. On the basis of restoration of milk parameters and standard plate count, the most effective therapy was elicited by Eucalyptus oil 98%+ DMSO 2% as teat dip in combination with antibiotic amoxicillin sulbactam (G3).

Effect of Subclinical Mastitis Detected in the First Month of Lactation on the Reproductive Performance of Dairy Cows in Western Algeria

Background: The current study aimed to detect in the first month of lactation the prevalence and risk factors of subclinical mastitis (SCM) and its effects on reproductive performance in dairy cows in western Algeria. Methods: 530 cows from 18 herds were examined in the study. California Mastitis Test (CMT) was used to screen subclinical mastitis for individual cows and a score of two or more for any quarter without any clinical symptoms and abnormalities in milk was considered as positive for SCM. Milk samples positive for subclinical mastitis were subjected to bacteriological analysis. Result: The present study revealed a prevalence of subclinical mastitis of 34.90%, (185 of 530). The most common pathogen identified from CMT-positive milk samples was Staphylococcus aureus with 35.62%. Logistic regression analysis showed that parity, farming system and milking method affect significantly the prevalence of SCM. Subclinical mastitis during the first month of lactation affected significantly the reproductive performance by increasing calving to first service interval (CFSI), calving-to-conception interval (CCI)and number of services per conception (NSPC). The results of this study highlight the importance of mastitis control programs in dairy farms by showing that a high frequency of subclinical mastitis caused by Staphylococcus aureus may have a negative impact on the reproductive performance of dairy cows.

Comparative prevalence assessment of subclinical mastitis in two crossbred dairy cow herds using the California mastitis test

The study was conducted on two herds of Jersey crossbred dairy cows. Samples were taken from two dairy farms to conduct the California Mastitis Test (CMT) during milking. A total of 115 animals were sampled, with 65 animals from Farm 1 and 60 animals from Farm 2. Milk samples were collected from all four quarters of each cow. The study analyzed a total of 460 quarters from the collected milk samples. Among these quarters, 132 quarters (28.69%) were found to be infected. The prevalence of mastitis was 45% in Herd 1 and 25% in Herd 2. The study found that only one quarter of the udder was more affected in both herds. The CMT score (likely a measure of mastitis severity) increased during late lactation. Infected cows had an average milk yield/day of 3.6 liters. Inflammatory (within or under the mammary gland) infections were more common in the first and second parity cows (cows that had given birth to one or two calves, respectively). This could be ascribed to the fact that dairy cows in their starting lactations are more vulnerable to the environmental pathogens because they might not have built up their immunity yet. Overall, this study provides insights into the prevalence, severity, and patterns of mastitis in Jersey crossbred dairy cows located at two different farms. The study highlights factors such as the stage of lactation, parity, and herd differences that can influence the occurrence and severity of mastitis in these animals.

Comparative Profiling of Milk Somatic Cells Proteomes Revealed Key Players in Mammary Immune Mechanisms During Mastitis in Tropical Sahiwal (Bos indicus) Cows.

Bovine mastitis poses a significant economic burden on the dairy industry worldwide. This pioneering proteomic study conducted a comparative profiling of milk somatic cell (SC) proteins contributing to mammary immune defense during subclinical and clinical mastitis (CM) in Sahiwal (Bos indicus) cows. Based on California mastitis test (CMT) scores, milk SC counts, differential leukocyte counts (DLCs), and bacteriological culture results, quarter milk SC samples were categorized into healthy (H), subclinical mastitis (SCM), and CM groups. Comparative proteome profiling of milk SCs was done using a label-free liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) proteomic approach. The identified upregulated proteins in mastitis groups such as Vanin 2, Thioredoxin reductase-like selenoprotein T, Ceramidase, Lymphocyte antigen 75, Misshapen-like kinase 1 (MINK1), Thrombospondin 1, Macrophage scavenger receptor 1, Leupaxin, and Lipoamide acyltransferase, involved in immune responses. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed immune functions and pathways like antigen processing, complement cascades, extracellular matrix receptor interaction, efferocytosis, leukocyte migration, chemokine, peroxisome proliferator-activated receptors (PPARs), and transforming growth factor (TGF)-beta signaling. These findings provide essential information on proteomic profiling in milk SCs and contribute valuable insights into immune-related proteins regulated during mastitis in dairy cows. Further, validated proteins (Vanin 2, MINK1, and Thrombospondin 1) offer potential inflammatory biomarkers for early mastitis detection in dairy cows.

Thermographic assessment of mastitis progression in sahiwal cattle: Insights into the patterns in the natural course of infection

Mastitis is a global concern in the dairy sector, demanding innovative solutions for effective management for quality lifetime milk production. In this study, infrared thermography (IRT) as a non-invasive technology was integrated into routine farm activities for continuous health monitoring of animals. For 30 days, we systematically monitored the udder health status in 40 Sahiwal cows (160 quarters), employing IRT along with the California Mastitis Test (CMT). We also assessed somatic cell count (SCC), microbial identification, and milk quality parameters of representative samples. The thermal imaging data was analyzed, considering both backward propagation from the 0th day to the −10th day and forward propagation from the 0th day to the +10th day. Our findings revealed that on the 0th day, the mean temperatures of the udder surface skin temperature (USST) and teat skin surface temperature (TSST) exhibited differences (p < 0.05) between the quarters affected by sub-clinical mastitis (SCM) and clinical mastitis (CM) in comparison to the healthy quarters, with the highest degree of difference observed. The observed temperature differences between CM and SCM quarters compared to healthy ranged from 1.8 to 3.62 °C and 0.98 to 3.23 °C for USST, and from 1.68 to 3.16 °C and 0.56 to 2.32 °C for TSST, respectively. Furthermore, our observations indicated that both udder and teat quarters responded differently to mastitis. A temperature rise of 1.37 °C in SCM quarters and 1.75 °C in CM quarters was observed between the −10th and -8th day relative to day 0, with the increase being more pronounced in the morning hours. Also, a notable temperature surge occurred during the -2nd and -1st days relative to the 0th day. The log10SCC values and milk quality parameters significantly differed (p < 0.05) between mastitis-affected and healthy samples. In addition, Staphylococcus spp. was identified as the predominant mastitis-causing pathogen in the bacteriological identification conducted in this study. Therefore, IRT efficiently assesses the initiation point of udder infection in Sahiwal cows, aiding in effective udder health management.

Distribution of Bovine Mastitis Pathogens in Quarter Milk Samples from Bavaria, Southern Germany, between 2014 and 2023-A Retrospective Study.

The objective of this study was to investigate the distribution of mastitis pathogens in quarter milk samples (QMSs) submitted to the laboratory of the Bavarian Animal Health Service (TGD) between 2014 and 2023 in general, in relation to the clinical status of the quarters, and to analyze seasonal differences in the detection risk. Each QMS sent to the TGD during this period was analyzed and tested using the California Mastitis Test (CMT). Depending on the result, QMSs were classified as CMT-negative, subclinical, or clinical if the milk character showed abnormalities. Mastitis pathogens were detected in 19% of the QMSs. Non-aureus staphylococci (NAS) were the most common species isolated from the culture positive samples (30%), followed by Staphylococcus (S.) aureus (19%), Streptococcus (Sc.) uberis (19%), and Sc. dysgalactiae (9%). In culture-positive QMSs from CMT-negative and subclinically affected quarters, the most frequently isolated pathogens were NAS (44% and 27%, respectively), followed by S. aureus (25% and 17%, respectively) and Sc. uberis (8% and 22%, respectively). In QMSs from clinically affected quarters, the most frequently isolated pathogens were Sc. uberis (32%), S. aureus (13%), Sc. dysgalactiae (11%), and Escherichia (E.) coli (11%). The distribution of NAS and Sc. uberis increased throughout the study period, while that of S. aureus decreased. From June to October, QMSs from subclinically affected quarters increased and environmental pathogens, such as Sc. uberis, were detected more frequently. In conclusion, this study highlights the dynamic nature of the distribution of mastitis pathogens, influenced by mastitis status and seasonal factors. Environmental pathogens still play an important role, especially in clinical mastitis and seasonal dependency, with the number of positive samples continuing to increase. It is therefore essential to continue mastitis control measures and to regularly monitor the spread of mastitis pathogens in order to track trends and adapt targeted prevention measures.

Effects of subclinical mastitis on automatic milking system data, hematological and biochemical parameters, and milk composition in Holstein cows.

Subclinical mastitis decreases milk production and quality, despite the normal appearance of the mammary glands and milk. Herein, we aimed to investigate changes in factors monitored via automatic milking systems (AMS) prior to subclinical mastitis onset and identify differences in hematological and biochemical parameters and milk composition at subclinical mastitis onset. Thirty-two Holstein cows were divided into two groups according to somatic cell counts (SCC) from AMS and milk composition analysis and the California mastitis test (CMT): healthy cows (controls [CON], n=16, SCC <500×103 cells/ml and negative for CMT) and cows with subclinical mastitis (SCM, n=16, SCC ≥500×103 cells/ml and positive for CMT). Eventually, 121 milk samples from the CON ([mCON], n=60) and SCM ([mSCM], n=61) groups were obtained; SCM samples were categorized as those from non-inflamed (mNQ) or subclinically-inflamed (mIQ) quarters. We evaluated AMS factors; hematological, biochemical, and milk composition parameters; and bacterial isolation. In cows with SCM, milk yield decreased and electrical conductivity (EC) changed before disease onset. Milk EC decreased in mNQ although increased in mIQ (p<0.05). The SCM group had higher globulin levels and lower basophil counts; albumin-to-globulin ratio; and total cholesterol, albumin, and blood urea nitrogen levels than the CON group (p<0.05). The mIQ group had higher SCC but lower levels of lactose and milk solids-not-fat than those in the mCON and mNQ groups (p<0.05). The mCON group had higher levels of milk non-protein nitrogen than the mNQ group (p<0.05). Opportunistic mastitis pathogens were isolated in the mIQ group. Changes in milk yield and EC measured using AMS occurred prior to subclinical mastitis, which may be associated with variation in basophil counts; albumin-to-globulin ratio; and total cholesterol, albumin, BUN, globulin, SCC, milk lactose, and milk solids-not-fat levels at disease onset. These findings provide new insights into early-stage subclinical mastitis.

Advancing mastitis assessment in dairy bovines via short milking tube thermography: A seasonal perspective.

In India, where dairy production leads globally, infrared thermography (IRT) and short milking tube thermography specifically are vital for managing mastitis. Therefore, the present study focuses on thermal imaging of the udder and short milking tube (SMT) of the milking machine during the peak milking process of Sahiwal cows and Murrah buffaloes during winter, summer, rainy and autumn seasons to identify sub-clinical (SCM) and clinical mastitis (CM) cases using the Darvi DTL007 camera. The udder health was assessed using the California Mastitis Test, Somatic Cell Count (SCC) and IRT throughout the year. Log10SCC and thermogram analysis revealed a difference (p < 0.01) between healthy, SCM, and CM cases during different seasons in both breeds. Further results showed an increase (p < 0.01) in SMT thermograms of SCM and CM cases compared to healthy quarters in Sahiwal cows during winter, summer, rainy, and autumn were 4.26 and 7.51, 2.37 and 4.47, 2.20 and 3.64, 2.90 and 4.94 ºC, respectively and for Murrah buffaloes were 3.56 and 5.55, 2.70 and 3.81, 1.72 and 3.10, 3.14 and 4.42ºC, respectively. The highest degree of increase in milking udder skin surface temperature and SMT of SCM and CM cases compared to healthy quarters was observed during the winter and the least during the rainy season. Thus, regardless of the seasons examined in this study, SMT thermograms effectively assessed SCM and CM.

A Review on Mastitis in Dairy Cows Research: Current Status and Future Perspectives

One of the most serious diseases affecting dairy cattle, causing significant losses both in breeding and economy, is mastitis, an inflammation of the mammary gland. Due to the economic importance of this issue, many research teams are striving to develop an easy-to-apply and, most importantly, effective method to prevent mastitis. The use of traditional methods for mastitis detecting and treating, as well as improvement in hygienic conditions, have not yielded the expected results in combating this disease combating. Currently, the main task is to find the tools that would allow for the rapid detection of mastitis and the improvement of udder health in cows while maintaining high milk production, which is essential for the profitability of dairy cattle farming. Accurate and rapid diagnostic tools, with the simultaneous capability of identifying pathogens, may help to reduce losses. Sufficient sensitivity and specificity for tests are required to minimize the number of false-positive and false-negative cases. Efforts are also being made to determine the optimal threshold value for detecting the disease at its earliest possible stage. The estimation of somatic cell count (SCC) as a phenotypic indicator of mastitis is widely used. A more precise parameter for accurately describing udder health is the differential somatic cell count (DSCC). The well-known California Mastitis Test (CMT) is an inexpensive, easy, and rapid method for mastitis detection useful on farms. The latest diagnostic methods for mastitis utilize tests based on the activity of N-acetyl-β-d-glucosaminidase (NAGase) or lactate dehydrogenase (LDH) as well as the determination of acute phase proteins (APPs) in blood serum and milk (such as haptoglobin, serum amyloid A, fibrinogen, and ceruloplasmin). Research also focuses on the genomic improvement of mastitis resistance in successive generations, and for this purpose, many quantitative trait loci (QTLs) and single nucleotide polymorphisms (SNPs) have been identified. In recent years, immunotherapy has become an increasingly common area of research, including vaccinations, T/B cell immunotherapy, RNA immunotherapy, epigenetic immunotherapy, stem cell therapy, and native secretory factors. An important aspect of the control of mastitis is the implementation of strategies that focus primarily on preventing the disease through appropriate breeding and farm management practices. In the forthcoming years, a significant challenge will be the development of universal diagnostic and therapeutic strategies that can be effectively implemented as alternatives to antibiotic therapy. Future research should prioritize the advancement of preventive and therapeutic techniques, such as immunotherapies, bacteriocins, herbal therapy, and nanoparticle technology.

Seasonal assessment of mastitis in crossbred goats: A thermographic approach

Sub-clinical mastitis exhibits a higher prevalence in dairy goats than clinical mastitis, necessitating the adoption of non-invasive diagnostic techniques such as infrared thermography (IRT) to detect this economically significant production disease in the dairy sector. Accordingly, this study aims to employ IR imaging of the udder and teat quarters of lactating crossbred goats (Alpine × Beetal and Sanen × Beetal) across various seasons, utilising IRT, to discern cases of sub-clinical (SCM) and clinical mastitis (CM). Over a year, 100–110 lactating crossbred dairy goats underwent consistent IRT screenings, followed by a comprehensive evaluation of udder health status using the California mastitis test and somatic cell count (SCC). The receiver operating characteristic (ROC) analysis was conducted to establish the cut-off values for different thermographic parameters in this study. The results revealed that the SCC increased significantly (p < 0.01) in healthy, SCM, and CM milk samples across the seasons. The analysis of ROC revealed a comparatively higher sensitivity, specificity, and accuracy for udder thermograms during SCM than CM and vice versa for teat thermograms. IRT analysis reflected a difference (p < 0.01) in the udder and teat thermograms among quarters of healthy, SCM, and CM in summer, winter, autumn, and rainy seasons. A significant increase (p < 0.01) in udder thermograms was observed for quarters affected with SCM and CM relative to healthy, with an increase of 1.89 and 2.94 °C in winter, 0.85 and 1.63 °C in summer, 0.73 and 1.41 °C in rainy, and 1.33 and 2.38 °C in autumn, respectively. Similarly, for teat thermograms it was 1.79 and 2.81 °C in winter, 0.76 and 1.41 °C in summer, 0.70 and 1.37 °C in rainy, and 1.09 and 1.93 °C in autumn, respectively. Therefore, regardless of the seasons examined in this study, IRT proves to be an effective and supportive tool for early mastitis detection in lactating crossbred dairy goats.

Genomic diversity and nutritional analysis of multi-drug resistant extended spectrum β-lactamase Producing-Klebsiella pneumoniae genes isolated from mastitic cattle milk in district peshawar, Pakistan

The increasing incidence of resistance extended spectrum-beta lactamase (ESBL) producing Klebsiella pneumonia become worldwide issue. The current study aimed to determine the genomic diversity of ESBL-producing K. pneumoniae in milk samples collected from cows with mastitis as well as their antibiotic sensitivity profiles and genetic identification in Peshawar, Pakistan. The california mastitis test (CMT) was initially used to verify the presence for mastitis in 700 collected milk samples. The molecular identification of the 16SrRNA gene confirmed 120/700 (17.14 %) propagation of K. pneumonia. Out of these isolates MDR ESBL-producing isolates were 60/120 (50 %). The lactose were found (M = 3.96 ± 0.28, SD = 2.19), followed by fats (M = 3.12 ± 0.11, SD = 0.90), protein (M = 5.97 ± 0.24, SD = 1.84), sodium (M = 55.74 ± 2.07, SD = 15.81), potassium (M = 138.5 ± 1.53, SD = 11.71), chloride (M = 0.74 ± 0.03, SD = 0.24), calcium (M = 10.27 ± 0.31, SD = 2.42), and chlorine (M = 2.80 ± 0.22, SD = 1.70), respectively. Amikacin (80 %), ceftazidime (71 %), and tetracycline (71 %) were shown to be the most effective antimicrobials against all of the isolates. The occurrence of the blaSHV gene was observed at 56.00 % whereas the blaTEM gene and blaCTX-M gene were 36.00 %, and 30.00 %. The distribution of blaCTX-M subgroup genes was followed by blaCTX-M-1 (38.00 %), blaCTX-M-9 (22.20 %), and blaCTX-M-15 (61.10 %). Co-occurrence of blaCTX-M+ blaSHV was (15.00 %), blaCTX-M+ blaTEM were (6.60 %), and blaSHV + blaTEM were (10.00 %), respectively. The inappropriate, prolonged and common use of antibiotics may apply selective pressure for propagation and the occurrence of resistant isolates.

The prevalence and factors associated with mastitis in dairy cows kept by small-scale farmers in Dodoma, Tanzania

Bovine mastitis is among the major diseases of economic importance in the dairy industry worldwide. Hygienic conditions during milking and housing are important determinants of mastitis infections. However, arid and semi-arid areas have a scarcity of water, which is an essential component of cleanliness. Thus, the study determined the prevalence of mastitis and associated factors in Dodoma, a semi-arid region in Tanzania. Three hundred and sixty-eight lactating cows were selected randomly from 114 farms in the Dodoma urban district for mastitis determination based on clinical signs and the California Mastitis Test. About 59.8 % of the cows had mastitis, and the subclinical type dominated. Factors associated with mastitis prevalence were cleanliness (hand and udder washing before milking), farmers' awareness of mastitis, the cow's parity and lactation stage, and herd size. Milk yield was not associated with the mastitis status of the cow but was related to the breed of the cow, whether warm or cold water was used to wash the cow's udder, parity, and lactation stage. Improvement in education on the importance of hygiene is necessary, as 38.6 % of the farmers were unaware of mastitis. Further detailed studies on microbiology, such as bacteriologic culture and polymerase chain reaction, are recommended to formulate interventions.

Virulence and resistance profiling of Staphylococcus aureus isolated from subclinical bovine mastitis in the Pakistani Pothohar region

Mastitis is considered one of the most widespread infectious disease of cattle and buffaloes, affecting dairy herds. The current study aimed to characterize the Staphylococcus aureus isolates recovered from subclinical mastitis animals in Pothohar region of the country. A total of 278 milk samples from 17 different dairy farms around two districts of the Pothohar region, Islamabad and Rawalpindi, were collected and screened for sub clinical mastitis using California Mastitis Test. Positive milk samples were processed for isolation of Staphylococcus aureus using mannitol salt agar. The recovered isolates were analyzed for their antimicrobial susceptibility and virulence genes using disc diffusion and PCR respectively. 62.2% samples were positive for subclinical mastitis and in total 70 Staphylococcus aureus isolates were recovered. 21% of these isolates were determined to be methicillin resistant, carrying the mecA gene. S. aureus isolates recovered during the study were resistant to all first line therapeutic antibiotics and in total 52% isolates were multidrug resistant. SCCmec typing revealed MRSA SCCmec types IV and V, indicating potential community-acquired MRSA (CA-MRSA) transmission. Virulence profiling revealed high prevalence of key genes associated with adhesion, toxin production, and immune evasion, such as hla, hlb, clfA, clfB and cap5. Furthermore, the Panton-Valentine leukocidin (PVL) toxin, that is often associated with recurrent skin and soft tissue infections, was present in 5.7% of isolates. In conclusion, the increased prevalence of MRSA in bovine mastitis is highlighted by this study, which also reveals a variety of virulence factors in S. aureus and emphasizes the significance of appropriate antibiotic therapy in combating this economically burdensome disease.

Thermal Imaging and Dimensionality Reduction Techniques for Subclinical Mastitis Detection in Dairy Sheep

Subclinical mastitis is a common and economically significant disease that affects dairy sheep production. Thermal imaging presents a promising avenue for non-invasive detection, but existing methodologies often rely on simplistic temperature differentials, potentially leading to inaccurate assessments. This study proposes an advanced algorithmic approach integrating thermal imaging processing with statistical texture analysis and t-distributed stochastic neighbor embedding (t-SNE). Our method achieves a high classification accuracy of 84% using the support vector machines (SVM) algorithm. Furthermore, we introduce another commonly employed evaluation metric, correlating thermal images with commercial California mastitis test (CMT) results after establishing threshold conditions on statistical features, yielding a sensitivity (the true positive rate) of 80% and a specificity (the true negative rate) of 92.5%. The evaluation metrics underscore the efficacy of our approach in detecting subclinical mastitis in dairy sheep, offering a robust tool for improved management practices.

MILK QUALITY OF ETAWA BREEDING GOATS SEEN FROM CALIFORNIA MASTITIS TEST, ALCOHOL TEST, REDUCTASE TEST, AND BOILING TEST IN RAKA ETAWA FARMING

Milk is a food product of animal origin that has high benefits. The main requirement for milk consumption by the public is good milk quality. This study aims to determine the quality of Etawa breeding goat milk as seen from the California mastitis test, alcohol test, reductase test, and boiling test at Raka Etawa Farm and see the relationship between the California mastitis test (CMT) with the alcohol test, reductase, and boiling test. This study was conducted in a descriptive observational. Sampling using purposive sampling method with sample criteria taken is milk from goat mothers who are in the lactation period. In this study it was found that of the 30 milk samples taken, based on the results of the California mastitis test, there were 15 samples of milk of Etawa breeding goats (50%) experiencing the incidence of subclinical mastitis level III, 11 milk samples (36,7%) experiencing the incidence of subclinical mastitis level II, and as many as 4 milk samples (13,3%) did not experience the incidence of subclinical mastitis. Based on the alcohol test, 26 milk samples (80%) showed positive results and as many as 4 milk samples (20%) showed negative results. The reductase test results showed that 24 samples (80%) had a reductase time of 2 hours and 6 samples (20%) showed a reductase time of 1 hour. The boiling test results showed that 26 milk samples (86,7%) were positive and 4 milk samples (13,3%) had negative results. There was no significant relationship between the CMT test and alcohol test and the CMT test and reductase test (P &gt; 0.05). There was a significant relationship between the CMT test and boiling test (P &lt; 0.05) on the quality of milk of Etawa breeding goats at Raka Etawa Farm. It can be concluded that the majority of Etawa breeding goat milk at Raka Etawa Farm is of poor quality. Cage hygiene, cleaning the udder before milking, and always using gloves during milking need to be considered to produce good quality milk by applicable standards.

Occurrence of antimicrobial-resistant bovine mastitis bacteria in Sakon Nakhon, Thailand.

Bovine mastitis is an inflammation of the mammary gland of dairy cattle that causes economic losses due to poor quantity and quality of milk. The extensive or incorrect use of antibiotics has increased in the veterinary field, leading to the emergence of antibiotic-resistant pathogens worldwide. This study aimed to investigate bovine mastitis bacterial pathogens in Sakon Nakhon, Thailand. A total of 35 dairy farms were screened for clinical and subclinical mastitis using the California Mastitis Test and clinical examination. Polymerase chain reaction was used to characterize bacterial species-induced mastitis (380 isolates) in cattle and antimicrobial resistance genes, and disk diffusion and broth microdilution were used to characterize antimicrobial susceptibility. The prevalence of Staphylococcus epidermidis (38.10%; 32/84)-induced mastitis in cattle was considerably high, followed by Streptococcus agalactiae (33.33%), Streptococcus uberis (25%), Klebsiella pneumoniae (8.33%), and Staphylococcus aureus (4.76%). In this study, Staphylococcus spp. isolates demonstrated 100% susceptibility to cefoxitin, and no antibiotic-resistance genes were identified. Tetracycline (TET) and macrolide-resistant genes of Streptococcus spp. revealed that tetM was predominant in 55.63% (79/142), followed by tetS + erm(B) (16.90%). Antibiotic susceptibility tests revealed the following resistance profiles to bacterial species: TET (85.92%), clindamycin (29.58%), erythromycin (15.49%), levofloxacin (14.08%), and penicillin (0%). Gram-negative bacterial isolates (K. pneumoniae [8.33%], Klebsiella variicola [2.38%], Klebsiella quasipneumoniae [1.19%], and Escherichia coli [1.19%]) were recovered and still susceptible to meropenem (100%), ceftazidime (97.06%), ceftriaxone (79.41%), and ciprofloxacin (79.41%). This result suggested that mastitis pathogens in this area were susceptible to most antimicrobials, with the exception of streptococci against TET. In this study, limited data were available including one from small-holder dairy farms and study only dairy farms in Sakon Nakhon, Thailand. So, more farms should be included in the future studies.

Antimicrobial resistance of Streptococcus dysgalactiae, Streptococcus agalactiae, and Streptococcus canis in quarter milk samples from Bavaria, Southern Germany, between 2012 and 2022

The objective of this study was to analyze the in vitro antimicrobial resistance (AMR) of Streptococcus (Sc.) dysgalactiae, Sc. agalactiae, and Sc. canis over a 10-year period from 2012 to 2022 against the most commonly used antimicrobial agents. For this purpose, all quarter milk samples (QMS) submitted to the milk laboratory of the Bavarian Animal Health Service (TGD) were analyzed. Each QMS was tested using the California Mastitis Test (CMT) and categorized as negative (N), subclinical (S), or clinical (C) mastitis if the milk character was abnormal. Samples with Sc. dysgalactiae, Sc. agalactiae, or Sc. canis were included and a subset of isolates were further tested for in vitro antimicrobial resistance by breakpoint analysis with broth microdilution. Sc. dysgalactiae (61%, n = 65,750) was the most abundant pathogen among those 3 species, followed by Sc. agalactiae (28%, n = 30,486), and Sc. canis (11%, n = 11,336). All 3 species showed the highest resistance to the same 4 antimicrobial agents: erythromycin, marbofloxacin, pirlimycin, and cefalexin/kanamycin with varying degrees of resistance. Throughout the study period, Sc. dysgalactiae, Sc. agalactiae, and Sc. canis were largely susceptible to the remaining antimicrobial agents tested (penicillin, amoxicillin-clavulanate, oxacillin, cefazolin, cefoperazone, cefquinome). Only less than 14% of isolates of Sc. dysgalactiae and Sc. canis were resistant against any of the antimicrobials tested. Sc. agalactiae was the species with the highest percentage of resistant isolates. While the percentage of resistant isolates from Sc. canis and Sc. dysgalactiae decreased, the percentage of resistant Sc. agalactiae isolates increased since 2017. In summary, most isolates were not resistant to the most commonly used antimicrobial agents for mastitis therapy, including β-lactam antibiotics and penicillin should remain the first-choice therapy against streptococcal mastitis.