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From 47 dairy farms located in the Anseba Region of Eritrea, 222 cows were randomly selected for a questionnaire survey, examined for signs of clinical mastitis and milk samples tested for subclinical mastitis using the California mastitis test (CMT). Twenty cows (9%) were deemed to have clinical mastitis (CM), because they had visible clinical signs or visible changes in milk samples. Of the 202 cows tested by the CMT, 156 (77.23%) had a score of 1 or above at least in one quarter and were deemed to have subclinical mastitis (SCM). SCM was significantly more frequent in exotic breeds than in cross or indigenous breeds ( p  < 0.001) and cows with poor body condition compared to cows with good body condition ( p  = 0.025). CM was significantly more frequent in cows with a history of clinical mastitis ( p  = 0.004). SCM was significantly more frequent in farms that keep dung inside the compound ( p  = 0.004), do not milk cows with mastitis last ( p  < 0.01), use earthen floor ( p  < 0.01), do not routinely use mastitis test ( p  < 0.001), have intensive system of production and in farms with frequent tick infestation of the udder ( p  < 0.001). From 20 CM-positive and 153-SCM-positive composite milk samples tested, 29 isolates of Staphylococcus aureus , 23 Enterococcus faecium and 20 Streptococcus species, including 8 Streptococcus agalactiae were isolated. Of the 13 isolates of S. aureus tested, all were resistant to tetracycline, 11 to sulfonamides and 10 to penicillin. Two Streptococcus agalactiae isolates tested were resistant to tetracycline and penicillin.

Somatic cell count (SCC) is one of the most important and widely used mastitis diagnostics. For detecting (sub)clinical mastitis, online SCC related measurements are more and more used in automatic milking systems (AMS). Sensors such as an automated online California Mastitis Test (O-CMT) allow for high frequency screening of high SCC cows within a herd, which makes it potentially powerful to identify episodes of mastitis. However, the performance of O-CMT measurements, as compared to SCC determined in the laboratory (L-SCC), has only scarcely been described. The aims of this study were (1) to assess the agreement between the O-CMT measurement averaged over different time windows and the corresponding L-SCC measurements; (2) to determine the optimal time window for averaging O-CMT as compared to L-SCC; (3) to explore the added value of time-series of frequent O-CMT measurements in individual cow udder health monitoring compared to L-SCC measurements. Data were collected from 50 farms in 6 different countries that were equipped with AMS using O-CMT measurements and also performed regular L-SCC testing. We found that the overall concordance correlation coefficient (CCC) between O-CMT and L-SCC was 0.53 but differed substantially between farms. The CCC between O-CMT and L-SCC improved when averaging O-CMT over multiple milkings, with an optimal time-window of 24 h. Exploration of time series of daily O-CMT recordings show that this is an effective screening tool to find episodes of high SCC. Altogether, we conclude that although O-CMT agrees moderately with L-SCC, because of its high measurement frequency, it is a promising on-farm tool for udder health monitoring.

Background Mastitis is a disease of major economic importance in dairy industry worldwide. It is of particular concern in developing countries like Ethiopia, where milk and milk products are scarce. The objectives of the study were to estimate the prevalence of mastitis, identify the cow-and herd-level potential risk factors and isolate Staphylococcus aureus , one of etiological agents for contagious mastitis, from cows positive for mastitis. A total of 529 lactating cows selected randomly from 95 herds were screened by California mastitis test (CMT) for sub-clinical mastitis. Also 172 milk samples collected from CMT positive cows were cultured for isolation of S. aureus . Results Based on CMT result and clinical examination, the prevalence of mastitis at herd-level was 74.7% (95% CI: 64.5, 82.8). The corresponding cow-level prevalence was 62.6% (95% CI: 58.3, 66.7), of which 59.2 and 3.4% were sub-clinical and clinical mastitis cases, respectively. S. aureus was isolated from 51.2% of the milk samples cultured and 73.2% of the herds affected with mastitis. In the multivariable logistic regression model, the herd-level factors significantly associated ( p  < 0.05) with the presence of mastitis were herd size, bedding material, and milking mastitic cows last, while at cow-level, breed, parity, stage of lactation, udder and leg hygiene, and teat end shape were noted to have a significant effect on mastitis occurrence. Conclusion The very high prevalence of mastitis, more importantly the sub-clinical one, in the herds examined revealed the huge potential economic loss the sector suffers. Perhaps this was attributed to lack of implementation of the routine mastitis prevention and control practices by all of the herd owners. The findings of this study warrants the need for strategic approach including dairy extension that focus on enhancing dairy farmers’ awareness and practice of hygienic milking, regular screening for sub-clinical mastitis, dry cow therapy and culling of chronically infected cows.

Subclinical mastitis (SCM) poses a significant threat to the global dairy industry, particularly in Bangladesh, where it remains a major constraint in buffalo dairy farming. The rising prevalence of antimicrobial resistant pathogens complicates disease management, resulting reduced milk yield, increased veterinary expenses, compromised animal welfare and potential risk to public health. This study investigated the prevalence and resistance profiles of multidrug-resistant (MDR) and extensively drug-resistant (XDR) Staphylococcus and Streptococcus species in raw buffalo milk from SCM cases in Bangladesh. A total of 1,540 quarter milk samples from 385 buffaloes were analyzed, revealing SCM prevalence rates of 67.9% (1046/1540; 95% CI: 65.6–70.3) at the quarter level and 80.8% (311/385; 95% CI: 76.5–84.6) at the animal level. Notable regional variations were observed, with Gowainghat showing the highest prevalence (88.1%; 141/160). This study did not identify any biologically plausible risk factors for the occurrence of SCM in buffalo. The Modified Whiteside Test and California Mastitis Test confirmed SCM, with culture and biochemical tests identifying 789 (51.2%) Staphylococcus spp. and 424 (27.5%) Streptococcus spp. isolates. Polymerase Chain Reaction (PCR) analysis indicated that 72.7% (456/627) of Staphylococcus isolates were Staphylococcus aureus , while the predominant Streptococcus species included Streptococcus uberis (32.3%) and Streptococcus dysgalactiae (14.9%). Resistance gene detection revealed a high prevalence of antimicrobial resistant genes (ARGs), particularly aac-3(iv) and tetA, across different buffalo quarters and habitats. Antibiogram profiling demonstrated high susceptibility to tetracycline (80.9; 83.1) and trimethoprim-sulfamethoxazole (87.4; 81.9), while significant resistance was noted against ampicillin (88.8; 87.1) and nalidixic acid (68.1; 62.1). MDR was observed in 76.4% (479/627) of Staphylococcus spp. and 67.3% (167/248) of Streptococcus spp. isolates, with 10.37% (65/627) and 10.48% (26/248) classified as possible XDR, respectively. These findings explored high antimicrobial resistance level among Staphylococcus and Streptococcus species in subclinical mastitis, highlighting the need for improved management practices and surveillance to mitigate public health risks posed by contaminated milk.

Dromedary camels are susceptible to mastitis, a multifactorial disease affecting dairy animals worldwide and leading to significant economic losses, particularly due to its subclinical form. However, limited data exist on the prevalence and risk factors associated with lactating camel mastitis in Egypt. Therefore, a cross-sectional study was conducted across three Egyptian governorates to assess the prevalence and identify potential risk factors. A total of 390 lactating camels were examined for both clinical and subclinical mastitis using the California Mastitis Test (CMT). The overall prevalence of mastitis was 38.46% (150/390), comprising 6.4% (25/390) clinical and 32.1% (125/390) subclinical cases. Significant associations (p < 0.05) were observed between mastitis prevalence and factors such as age, lactation stage, tick infestation, milking hygiene, and the presence of udder or teat lesions. The likelihood of mastitis was six times higher in camels older than five years, three times higher during early lactation, twice as high in the presence of tick infestation, twice as high with poor milking hygiene, and three times higher in animals with udder or teat lesions. Among the 600 quarter milk samples obtained from positive animals, mastitis-causing pathogens were isolated from 380 samples (63.3%), while 220 samples (36.7%) showed no bacterial growth. Streptococcus spp. (excluding S. agalactiae ) and Escherichia coli were the most prevalent isolates (26.1% and 25%, respectively), whereas S. agalactiae was the least frequent, detected in only 5.5% of the samples. These findings highlight the importance of implementing integrated control measures to reduce mastitis prevalence, enhance camel milk quality, mitigate economic losses, and safeguard public health.

This study aimed to isolate and characterize Staphylococcus aureus (S . aureus) from samples of mastitis milk taken from dairy cattle in Punjab’s Narowal District. 200 milk samples were collected aseptically from different dairy herds in the district, including clinical mastitis (CM) and sub-clinical mastitis (SCM) cows. Blood agar and mannitol salt agar were used for S . aureus isolation and identification. Selective media were then used for additional purification. Observations of morphological and biochemical traits verified the existence of S . aureus . Through questionnaire-based surveys, the prevalence of S . aureus mastitis was identified, and risk variables linked to its incidence were evaluated. The findings showed that S . aureus mastitis was prevalent in 42.5% of cases, with sub-clinical cases having a greater incidence (45.8%) than clinical cases (37.5%). Risk factors such as distance from dung pile to stall(m), source of water, dung removal per day, lactation period (weeks), parity, effect of milk yield (L), traumatic injury of udder, housing type, floor type, teat dipping, and bedding type was identified and their relationship to the occurrence of S . aureus mastitis was studied. Tests for antibiotic sensitivity revealed that S . aureus isolates were responsive to gentamycin, ceftiofur, tetracycline, enrofloxacin, and ciprofloxacin but extremely resistant to penicillin and amoxicillin. Additionally, the Somatic Cell Count (SCC) and California Mastitis Test (CMT) were used at different intervals to assess the effectiveness of the medication. Furthermore, compared to other treatment groups, a larger percentage of cure rates was seen in the groups receiving ceftiofur and enrofloxacin. Overall, this study contributes to the development of more effective management methods for S . aureus mastitis in dairy calves by offering insightful information about the condition’s prevalence, risk factors, antibiotic sensitivity, and effectiveness of treatment.

This study aimed to characterize virulence and antibiotic resistance genes in multidrug-resistant (MDR) and extensively drug-resistant (XDR) Escherichia coli and Klebsiella pneumoniae isolated from cases of subclinical mastitis (SCM) in buffaloes. A cross-sectional study was conducted on 1540 quarter milk samples collected from 385 buffaloes. Milk samples were screened using the California Mastitis Test and Modified Whiteside Test. Positive samples underwent bacterial culture, biochemical tests, biofilm detection and molecular analysis for pathogen identification and detection of virulence, resistance, and extended-spectrum beta-lactamase (ESBL) genes. The prevalence of SCM was 67.9% (1046/1540) at the quarter level and 80.8% (311/385) at the animal level. E. coli was identified in 9.5% (146/1540) of the samples, while K. pneumoniae was detected in 9.09% (140/1540). Virulence genes, such as stx1 (27.4%), and resistance genes, including aac(3)-iv (77.4%) and tetA (76.7%), exhibited higher prevalence. Additionally, β-lactamase genes, notably bla (67.1%), and ESBL genes, such as bla , were detected. Biofilm formation was detected in 83.6% (122/146) of E. coli isolates and 75.7% (106/140) of K. pneumoniae isolates. Antimicrobial susceptibility testing revealed significant resistance to ampicillin, amoxicillin-clavulanic acid, and aminoglycosides. MDR was observed in 31.5% of E. coli and 39.3% of K. pneumoniae isolates, with XDR rates of 8.9% and 12.9%, respectively. These findings underscore the alarming spread of resistant pathogens in SCM-affected buffaloes, emphasizing the urgent need for ongoing surveillance and targeted intervention strategies.

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.

Methicillin-resistant Staphylococcus aureus (MRSA) is a critical pathogen implicated in subclinical mastitis (SCM), a hidden threat to dairy productivity. This study investigated the prevalence, antibiotic resistance profiles, and virulence traits of MRSA from SCM-affected riverine buffaloes in Jamalpur, Bangladesh. A total of 344 milk samples were screened using the California Mastitis Test (CMT) and Modified Whiteside Test (MWST). Among the milk samples, 46.5% were positive for SCM by CMT. Culture, biochemical tests, and PCR confirmed 73 (21.2%) Staphylococcus spp., of which 30 (41.1%) were identified as S. aureus and 43 (58.9%) as non-aureus staphylococci (NAS). Among the 30 S. aureus -positive isolates, 10 (33.3%) were identified as methicillin-resistant S. aureus (MRSA), corresponding to a prevalence of 2.9% among the total milk samples. The MRSA isolates exhibited high multidrug resistance, especially to tetracycline (80%) and cefoxitin (80%), and commonly harbored resistance genes such as tetA (80%), aac(3)-iv (70%), and sul1 (50%). Virulence genes hla (66.7%) and sea (50%) were frequently detected, while icaA was found in 23.3% of MRSA. Notably, 60% of MRSA isolates were categorized as XDR based on international standard definitions, while 60% were biofilm producers with high MARI values up to 0.92, indicating severe resistance potential. These findings underscore a significant burden of MDR/XDR MRSA with virulence potential in buffalo SCM, posing serious risks to animal and public health.

MicroRNA (miRNA) in tissue and liquid samples have been shown to be associated with many diseases including inflammation. We aimed to identify inflammation-related miRNA expression level in the bovine mastitis milk. Expression level of inflammation-related miRNA in milk from mastitis-affected and normal cows was analyzed using qPCR. We found that expression level of miR-21, miR-146a, miR-155, miR-222, and miR-383 was significantly upregulated in California mastitis test positive (CMT+) milk. We further analyzed these miRNA using a chip-based QuantStudio Digital PCR System. The digital PCR results correlated with those of qPCR, demonstrating upregulation of miR-21, miR-146a, miR-155, miR-222, and miR-383 in CMT+ milk. In conclusion, we identified miRNA that are upregulated in CMT+ milk. These miRNA exhibited sensitivity and specificity greater than 80% for differentiating between CMT+ milk and normal milk. Our findings suggest that inflammation-related miRNA expression level in the bovine milk was affected by mastitis, and miRNA in milk have potential for use as biomarkers of bovine mastitis.