Poor mastitis control favors intramammary infection (IMI), which always involves CNS. This study aimed to determine the relationships of IL-4, IL-6, and IL-10 in mastitis milk with concurrent infection, bacterial pathogens, SCC, and MDA, an oxidative stress marker. All mastitis quarters from five smallholder dairy farms were sampled aseptically before morning milking and again before afternoon milking for bacteriological identification using MALDI-TOF mass spectrometry. The samples with the concomitant infection between streptococci and CNS and their pairs of another sample from the quarters were selected. In addition, samples were randomly chosen to have a controlled single infection. IL-4, IL-6, and IL-10 were measured with ELISA kits. MDA was measured using HPLC, while SCC was measured using Fossomatic™ FC. The results from a repeated measure analysis showed that IL-4 positively correlated with SCC, while IL-6 showed a negative trend. IL-4 levels were highest in CNS infections and significantly higher than in non-infected or mixed infections (p < 0.05). The IL-6 level of the mixed bacteria was highest and showed a different trend from non-infection, and the quarter was infected with streptococcal bacteria. In conclusion, from a single infection, the streptococci and CNS quarter showed varied immune responses, including trendily higher IL-6 and IL-4.

The microbial ecology in mastitis involves the interactions between bacteria and the mammary gland environment. Poor mastitis control, for which understanding these microbial relationships is crucial, increases the risk of mastitis and co-infections. The aim of this study was to determine the pathogenesis and bacterial ecology of murine mammary glands following intramammary infection (IMI) with S. aureus (AU), S. agalactiae (SA), and four isolates of selected non-aureus staphylococci (NAS), as well as co-infections of AU or SA with NAS. Selected NAS demonstrated resistance to growth inhibition after co-culture with AU and SA and were proven to be protective in vitro against AU and SA. Both single infections and co-infections of AU, SA, two selected isolates of S. hominis (NAS1 and NAS2), and two selected isolates of S. chromogens (NAS3 and NAS4) were injected into the murine mammary glands at 105 CFU/mL. After 72 h of inoculation, the results showed that AU, AU–NAS2, and AU–NAS3 expressed severe inflammation with clinical signs of mastitis. The persistence of AU was found for AU, NAS3, and AU–NAS3, whereas the persistence of SA was found for SA–NAS1, SA–NAS3, and SA–NAS, although single SA could not persist. Interestingly, NAS2 in both the single- and co-IMIs with AU or SA resulted in a spontaneous cure 72 h after the IMI. In conclusion, some NAS have beneficial results because they can eliminate S. aureus and S. agalactiae, but some NAS have negative effects because they can support persistent S. agalactiae infection. These results may be used to explain both the advantages and disadvantages of NAS on farms with poor mastitis control. In addition, the beneficial S. hominis may initiate natural live antibiotics and reduce antibiotic resistance problems.

Bovine mastitis is a major problem affecting dairy production, and teat disinfection is important for reducing the risk of bacterial infection. Curcumin has antimicrobial properties, but its poor water solubility limits its use in aqueous formulations. This study aimed to develop chlorhexidine–curcumin nanoformulations (CCNs) and perform preliminary in vitro screening of these nanoformulations as candidate teat-dipping formulations. Four CCNs containing increasing concentrations of curcumin (0%, 0.1%, 0.5%, and 0.9% w/v; CCN0, CCN0.1, CCN0.5, and CCN0.9, respectively) were prepared. CCN0, CCN0.1, and CCN0.5 were formulated as oil-in-water nanoemulsions, whereas CCN0.9 was prepared as a curcumin-β-cyclodextrin inclusion complex. The physicochemical properties were characterized, and the antibacterial activity was evaluated in vitro by agar well diffusion against six mastitis-associated pathogens. CCN0.9 showed the largest inhibition zones overall (20.07 ± 1.05–28.85 ± 1.05 mm; p < 0.05), while CCN0.1 and CCN0.5 showed activity comparable to or greater than that of commercial teat dips against several pathogens. CCN0.5 showed the most favorable nanoemulsion characteristics, with a particle size of 94.77 nm and a zeta potential of −46.57 mV. Overall, CCNs demonstrated antibacterial activity against mastitis pathogens, supporting their potential as candidate teat-dip formulations. Further teat irritation and in vivo efficacy studies are needed before field application.

Early detection of canine cognitive dysfunction syndrome (CDS) is challenged by the limitations of single diagnostic tools, including owner-reported subjectivity and impractical behavioral testing protocols. This study examined a multidomain strategy integrating questionnaires, behavioral testing, and a circulating biomarker to identify measures associated with mild cognitive impairment in aged dogs. A total of 21 aged dogs were classified as cognitively normal (n = 9) or mildly cognitively impaired (n = 12) using the Canine Dementia Scale (CADES), and they were further assessed based on the Canine Cognitive Dysfunction Rating Scale (CCDR), a battery of behavioral tests across six cognitive domains, and serum brain-derived neurotrophic factor (BDNF) concentrations. The CADES scores differed significantly between the mildly cognitively impaired and cognitively normal groups (median 12.5 vs. 0, p < 0.01), whereas the CCDR scores did not (p = 0.114). Among the behavioral measures, the delayed match-to-sample task with a 20-s delay and the recognition index derived from novel object recognition testing were significantly associated with cognitive group classification; the other domains showed no significant associations. Serum BDNF concentrations did not differ between the groups or predict group membership. The results of this exploratory study show that targeted behavioral measures may be more strongly associated with mild cognitive impairment than circulating BDNF, thus supporting their potential utility for early cognitive screening in clinical settings. However, these findings should be interpreted with caution, and their diagnostic accuracy should be confirmed in larger cohorts with further validation.