Gold Nanoparticle-Based Plasmonic Detection of Escherichia coli, Salmonella enterica, Campylobacter jejuni, and Listeria monocytogenes from Bovine Fecal Samples.

Current diagnostic methods for detecting foodborne pathogens are time-consuming, require sophisticated equipment, and have a low specificity and sensitivity. Magnetic nanoparticles (MNPs) and plasmonic/colorimetric biosensors like gold nanoparticles (GNPs) are cost-effective, high-throughput, precise, and rapid. This study aimed to validate the use of MNPs and GNPs for the early detection of Escherichia coli O157:H7, Salmonella enterica spp., Campylobacter jejuni, and Listeria monocytogenes in bovine fecal samples. The capture efficiency (CE) of the MNPs was determined by using Salmonella Typhimurium (ATCC_13311) adjusted at an original concentration of 1.5 × 108 CFU/mL. One (1) mL of this bacterial suspension was spiked into bovine fecal suspension (1 g of fecal sample in 9 mL PBS) and serially diluted ten-fold. DNA was extracted from Salmonella Typhimurium to determine the analytical specificity and sensitivity/LOD of the GNPs. The results showed that the CE of the MNPs ranged from 99% to 100% and could capture as little as 1 CFU/mL. The LOD of the GNPs biosensor was 2.9 µg/µL. The GNPs biosensor was also tested on DNA from 38 naturally obtained bovine fecal samples. Out of the 38 fecal samples tested, 81.6% (31/38) were positive for Salmonella enterica spp., 65.8% (25/38) for C. jejuni, 55.3% (21/38) for L. monocytogenes, and 50% (19/38) for E. coli O157:H7. We have demonstrated that MNP and GNP biosensors can detect pathogens or their DNA at low concentrations. Ensuring food safety throughout the supply chain is paramount, given that these pathogens may be present in cattle feces and contaminate beef during slaughter.

Epidemiological Survey of Peste des Petits Ruminants in Ethiopia: Cattle as Potential Sentinel for Surveillance.

Peste des petits ruminants (PPR) is a highly contagious viral disease of small ruminants; it emerged in countries previously free of the disease following the eradication of rinderpest. PPR is classified by international organizations as the next priority animal disease for global eradication campaign. Assessment of the local situations is the first step in the eradication efforts. The objective of this study was to investigate and compare the seroprevalence of PPR in cattle, sheep, and goats under two livestock production systems in Ethiopia: North Shewa zone of Amhara region represents a highland sedentary life style characterized by mixed livestock-crop production system; Zone Three of Afar region represents a lowland nomadic life style characterized by pastoral livestock production system. N-competitive ELISA PPR test was performed on sera from 2,993 animals ≥6 months old sampled at watering and grazing points. Multivariable logistic regression models comparing the seropositivity between the two production systems were built by classifying doubtful results as positive, negative, or excluding them from the data. The odds ratio (OR) comparing overall PPR seroprevalence in the sedentary North Shewa Zone compared to the nomadic Zone Three ranged from 19 to 27 (P < 0.001), depending on how doubtful results were classified, which contrasts with what has been reported in the literature. This is not likely to be related solely to vaccination, since seroprevalences in cattle and small ruminants were similarly high or low in the respective zones (0-4% for Zone Three and 20-40% for North Shewa Zone), and cattle were not likely to be vaccinated. The OR of seropositivity for goats compared to cattle ranged from 1.9 [95% confidence interval (CI): 1.3-2.7; P < 0.001] to 2.2 (95% CI: 1.5-3.1; P < 0.001) when doubtful results were excluded or classified as negative, respectively. When doubtful results were classified as positive, association between seropositivity and animal species was not significant (P > 0.05). Our results suggest to further investigate cattle as sentinel animals for PPR surveillance.

Quantitative risk assessment of entry of contagious bovine pleuropneumonia through live cattle imported from northwestern Ethiopia.

Contagious bovine pleuropneumonia (CBPP) is a highly contagious bacterial disease of cattle caused by Mycoplasma mycoides subspecies mycoides small colony (SC) bovine biotype (MmmSC). It has been eradicated from many countries; however, the disease persists in many parts of Africa and Asia. CBPP is one of the major trade-restricting diseases of cattle in Ethiopia. In this quantitative risk assessment the OIE concept of zoning was adopted to assess the entry of CBPP into an importing country when up to 280,000 live cattle are exported every year from the northwestern proposed disease free zone (DFZ) of Ethiopia. To estimate the level of risk, a six-tiered risk pathway (scenario tree) was developed, evidences collected and equations generated. The probability of occurrence of the hazard at each node was modelled as a probability distribution using Monte Carlo simulation (@RISK software) at 10,000 iterations to account for uncertainty and variability. The uncertainty and variability of data points surrounding the risk estimate were further quantified by sensitivity analysis. In this study a single animal destined for export from the northwestern DFZ of Ethiopia has a CBPP infection probability of 4.76×10(-6) (95% CI=7.25×10(-8) 1.92×10(-5)). The probability that at least one infected animal enters an importing country in one year is 0.53 (90% CI=0.042-0.97). The expected number of CBPP infected animals exported any given year is 1.28 (95% CI=0.021-5.42). According to the risk estimate, an average of 2.73×10(6) animals (90% CI=10,674-5.9×10(6)) must be exported to get the first infected case. By this account it would, on average, take 10.15 years (90% CI=0.24-23.18) for the first infected animal to be included in the consignment. Sensitivity analysis revealed that prevalence and vaccination had the highest impact on the uncertainty and variability of the overall risk.