Citrus-derived oil inhibits Staphylococcus aureus growth and alters its interactions with bovine mammary cells.

This experiment examined the effects of cold-pressed, terpeneless citrus-derived oil (CDO) on growth of Staphylococcus aureus, which a major cause of contagious bovine mastitis, and invasion of bovine mammary cells (MAC-T). To determine minimum inhibitory concentration, we used the broth dilution method, using CDO concentrations range from 0.0125 to 0.4% with 2-fold dilutions. Growth inhibition was examined by adding 0.00, 0.05, 0.025, 0.0125, and 0.00625% CDO to 10(5) cfu/mL S. aureus in nutrient broth and enumerating colonies after serial dilution. In a 96-well plate, S. aureus (10(7) cfu/mL) was allowed to form a biofilm, treated with 0, 0.025, 0.5, or 1% CDO, and then was measured using a spectrophotometer. Cytotoxic effect on immortalized MAC-T cells was also examined at various concentrations of CDO using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. We observed that the minimum inhibitory concentration of CDO to inhibit the growth of S. aureus in vitro was 0.025% CDO. A time kill curve for CDO action on S. aureus over 4h was generated. The CDO completely eliminated S. aureus after 3h of incubation at a concentration of 0.25%, or after 2h of incubation at concentrations of 0.05%. It was also observed that CDO had no effect on preformed biofilms except at a concentration of 0.05%, in which a significant reduction in the measured absorbance was noted. In addition, the association and invasion of S. aureus to MAC-T cells were significantly inhibited after 1h of treatment with CDO. Citrus-derived oil was also able to increase cellular proliferation of MAC-T cells at concentrations up 0.05% and had no effect at a concentration of 0.1% after 1 h. Our data suggests that CDO should be considered for further research as a preventive and therapeutic against bovine mastitis.

Zoonotic bacterial pathogens and mixed crop-livestock farming.

Use of mixed crop-livestock farms (MCLFs) is one of the oldest and most traditional farming methods practiced all over the world, and MCLFs are still one of the major systems of food production, particularly for organic foods. On these typically small farms, livestock are reared primarily on grass and naturally grown crops, while composted animal wastes are used to fertilize the soil for growing crops. Specific to organic MCLFs, biosecurity challenges arise from the fact that animals are reared outdoors, which increases potential for contact with disease vectors including wild birds, rodents, and insects. Organic regulations do not allow the use of chemicals and antibiotics; therefore, alternative methods for control of disease and zoonotic pathogens must be used. Due to the biosecurity challenges and the complexity of the MCLF environment, methods for control of zoonotic pathogens need to be carefully considered in order to be effective and to abide by organic regulations if required. The objectives of this study are to define the complex routes of transmission, as well as the prevalence of potential zoonotic and possible interruption strategies of these pathogens among the food animals and crops produced on MCLFs.

Inhibition of growth and alteration of host cell interactions of Pasteurella multocida with natural byproducts.

Pasteurella multocida is a leading cause of fowl cholera in both free-range pasture and conventional/commercially raised poultry. Its infection is a serious threat to poultry health and overall flock viability. Organic poultry is comparatively more vulnerable to this pathogen. It is a significant cause of production loss and price increase of poultry products, specifically organic poultry products. Some plant products are well documented as sources of natural antimicrobials such as polyphenols found in different berry pomaces and citrus oil. Pomace, a byproduct (primarily of seeds and skins) of fruits used for juice and wine production, and citrus oil, the byproduct of citrus juice production, show promising antimicrobial activity against various pathogens. Here, we showed for the first time that blackberry and blueberry pomace extracts and citrus oil inhibited P. multocida growth. Minimum bactericidal concentrations were determined as 0.3 and 0.4 mg/mL gallic acid equivalent for blackberry and blueberry pomace extracts, respectively. Similarly, only 0.05% citrus oil (vol/vol) completely inhibited P. multocida growth. Under shaking conditions, the antimicrobial activity of both pomace extracts and citrus oil was more intensive. Even citrus oil vapor also significantly reduced the growth of P. multocida. In addition, cell surface hydrophobicity of P. multocida was increased by 2- to 3-fold and its adherence to chicken fibroblast (DF1) and bovine mammary gland (MacT) cells was reduced significantly in the presence of pomace extracts only. This study indicates that these natural products might be good alternatives to conventional antimicrobial agents, and hence, may be used as feed or water supplements to control fowl cholera and reduce production loss caused by P. multocida.

Ecological Dynamics of Campylobacter in Integrated Mixed Crop-Livestock Farms and Its Prevalence and Survival Ability in Post-Harvest Products.

This study investigated the ecological distribution of zoonotic bacterial pathogen, Campylobacter, in mixed crop-livestock (MCL) farms compared to conventional farms and their products at pre- and post-harvest levels. A total of 222 Campylobacter isolates were identified. At pre-harvest level, a total of 1287 samples from seven MCL farms, four conventional poultry farms, four organic produce-only and five conventional produce-only farms from Maryland and the DC metropolitan area were analysed from 2012 to 2014. Campylobacter was detected in 11.16% and 3.6% of MCL and conventional farm samples, respectively, but none from produce-only farm samples. Tetracycline resistance was observed in 51.02% of MCL farm isolates but none among conventional farm isolates. For post-harvest analysis, a total of 1281 food products from seven farmers markets, three organic retail supermarkets and three conventional retail supermarkets were collected from the same area. Campylobacter was isolated in 87.5%, 71.43% and 33.33% of whole chicken carcasses in farmers markets, organic and conventional retail supermarkets, respectively. No Campylobacter was detected in post-harvest produce samples due in part to the inability of Campylobacter to survive in absence of sufficient water activity. Overall, this study reveals public health concerns regarding the MCL farm environment and their products that are sold in retail and farmers markets.