Leptospirosis is caused by pathogenic Leptospira, and synanthropic and wildlife species of rodents are an important source of infection; however, much of the information about infection progression was obtained from murine models. The aim of this study was to assess infection status and risk factors associated with pathogenic Leptospira in synanthropic and wild rodent species and describe histopathological lesions in several organs from naturally infected animals. In a cross-sectional study, 121 rodents from three synanthropic species and two wild species were trapped in dairy farms in Southern Chile. Liver, heart, kidney, and lungs from trapped animals were fixed in formalin and stained with hematoxylin–eosin. Tissues with lesions consistent with Leptospira infection were tested by immunohistochemistry (IHC) and real-time polymerase chain reaction (qPCR) using the LipL32 antigen. Risk factors were assessed by a conditional mixed-logistic regression model. More than half (56.7%) of the negative reactors to the microscopic agglutination test were identified as infected either by IHC/qPCR. A lower risk of infection compared to the rest of the seasons was found in the fall, and the synanthropic species have a lower risk of infection in comparison with the wildlife species. IHC and qPCR contributed to the identification of pathogenic Leptospira in related histological lesions and 50% more infections than serology.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of paratuberculosis, a contagious infectious disease that affects domestic and wild ruminants causing chronic inflammation of the intestine. MAP has proven to be very resistant to both physical and chemical processes, making it difficult to control this pathogen. Based on the recognized antimicrobial properties of copper, the objective of this study was to evaluate the effectiveness of copper ions to reduce MAP numbers and/or MAP viability in a fluid matrix. Besides, methicillin-resistant Staphylococcus aureus (MRSA), and Escherichia coli were used as controls of the effectiveness of copper ions. MAP-spiked PBS was subjected to copper ions treatment at 24 V for 5 min and the PBS suspensions were sampled before and after treatment. MAP viability and quantification were determined using three complementary techniques: a phage amplification assay, MGIT culture and qPCR. Moderate numbers (103 CFU ml−1) of the two control bacteria were completely eliminated by treatment with copper ions. For MAP, copper ions treatment reduced both the viability and numbers of this pathogen. Phage assay information quickly showed that copper ions (24 V for 5 min) resulted in a significant reduction in viable MAP. MGIT culture results over time showed statistically significant differences in time-to-detection (TTD) values between PRE and POST treatment. MAP genome equivalent estimates for PBS suspensions indicated that MAP numbers were lower in samples POST-treatment with copper ions than PRE-treatment. The use of copper ions resulted in a significant reduction of MAP in a liquid matrix, although some MAP survival on some occasions was observed.

One of the important routes of Mycobacterium avium subsp. paratuberculosis (MAP) transmission in dairy calves is milk. The aim of the present study was to assess the efficacy of milk treatments to prevent MAP infection transmission to calves. A one-year longitudinal study was carried out. Newborn calves were assigned to one of four experimental groups: 5 calves received naturally MAP-contaminated milk, 5 calves received copper treated milk, 4 calves were fed calf milk replacer, and 3 were fed UHT pasteurized milk. MAP load in milk was estimated. Infection progression was monitored monthly. After one year, calves were euthanized, and tissue samples were cultured and visually examined. MAP was undetectable in milk replacer and UHT milk. Copper ion treatment significantly reduced the number of viable MAP in naturally contaminated milk. Fecal shedding of MAP was observed in all study groups but began earlier in calves fed naturally contaminated milk. Paratuberculosis control programs must place multiple hurdles between the infection source, MAP-infected adult cows, and the most susceptible animals on the farm, young calves. As our study shows, strict dependence on a single intervention to block infection transmission, no matter how important, fails to control this insidious infection on dairy farms.

Background: Mycobacterium avium subsp. paratuberculosis (MAP) is the causal agent of paratuberculosis. This pathogen is able to survive adverse environmental conditions, including the pasteurization process. Copper, a well-studied metal, is considered an important antibacterial tool, since it has been shown to inactivate even MAP in treated milk through unknown mechanisms. The aim of the present study is to show the effect of copper ions, and reactive oxygen species (ROS) generated in response to oxidative stress, on the damage to MAP DNA when exposed to a copper ion challenge in cow’s milk. Methodology: Spiked milk with different MAP bacterial loads was supplemented with blocking agents. These were either the copper chelators ethylenediaminetetraacetic acid (EDTA) and batocuproin (BCS) or the ROS quenchers D-mannitol, gallic acid and quercetin. The DNA protection, MAP viability and ROS production generated after exposure to a copper challenge were then measured. Results: In a bacterial load of 10⁴ cells mL⁻¹, blocking effects by both the copper chelators and all the ROS quenchers offered significant protection to MAP DNA. In a concentration of 10² cells mL⁻¹, only D-mannitol and a mix of quenchers significantly protected the viability of the bacteria, and only at a concentration of 10⁶ cells mL⁻¹ was there a lower production of ROS when supplementing milk with gallic acid, quercetin and the mix of quenchers. Conclusion: Based on these findings, it may be concluded that MAP DNA damage can be attributed to the combined effect of the direct copper ions and ROS generated. Nevertheless, taking into account the antioxidant environment that milk provides, the direct effect of copper could play a prominent role.

Background/Objectives: Waste milk harbors many bacteria and antibiotic residues. Calves fed with untreated waste milk have a higher incidence of scours and an increased risk of developing antimicrobial-resistant bacteria. This study aimed to evaluate the efficacy of treatment with copper ions on bacteria and antibiotics contained in bovine waste milk. Methods: Waste milk samples were collected from a dairy farm for seven weeks and were subjected to treatment with copper ions. Total bacterial counts, coliforms, Streptococcus, and Staphylococcus were assessed before and after treatment. Additionally, metagenomic analysis was performed to determine microbial diversity. Results: Before treatment, the total bacterial count average was 4.0 × 10⁶ CFU/mL, 1.7 × 10⁴ CFU/mL for coliforms, 2.6 × 10⁶ CFU/mL for Streptococcus, and 5.4 × 10² CFU/mL for Staphylococcus Copper treatment significantly reduced bacterial counts within 15 min. Total bacteria decreased from 4.0 × 10⁶ CFU/mL to 1.1 × 10² CFU/mL after 30 min; meanwhile, other groups were not detected. The most abundant groups were Lactococcus (29.94%), Pseudomonas (28.89%), and Enterobacteriaceae (21.19%). Beta-lactams were detected in five-sevenths samples, and in one sample they were detected before and at 15 min of treatment but not after 30 min. Conclusions: The effect of treatment with copper ions on the different bacterial groups was significantly effective but showed limited effect on the detection of antibiotics.

Neonatal dairy calves are highly susceptible to Mycobacterium avium ssp. paratuberculosis (MAP) infection, but data remain limited on early infection prevalence and transmission drivers. This study aimed to estimate the true prevalence of MAP infection and identify associated risk factors in Chilean dairy calves younger than 60 d of age. Fecal and environmental samples were collected from 579 calves across 39 dairy herds. The MAP detection used a phage-based method (magnetic phage separation-quantitative PCR) that selectively identifies viable bacteria, and a Bayesian model was used to account for biased diagnostics test results. The overall estimated true prevalence of MAP infection was 4.4% (95% posterior probability intervals [PPI]: 0%-19.4%). Environmental contamination with MAP was frequently detected in calf pens (19.4% of pens; 30.7% of herds). Although multiple transmission routes exist, the presence of viable MAP in the calf's immediate pen environment was identified as the primary risk factor associated with infection in these neonatal calves (odds ratio: 3.7; 95% PPI: 1.2-9.0). These findings suggest that MAP infection could occur very early in life in this population, and that contamination within the calf's immediate environment is a key determinant of early-life MAP infection status. This highlights the critical need for implementing rigorous environmental hygiene control measures, specifically within calving and calf-rearing areas, starting immediately after birth, to effectively mitigate early MAP exposure and transmission in dairy herds.