Development of a qPCR for the detection and quantification of Salmonella spp. in sheep feces and tissues.

Salmonella spp. are common causes of disease in intensive livestock production systems, and contamination of foodstuffs is of significant concern for public health. Therefore, the identification and quantification of Salmonella spp. is important for monitoring the level of fecal shedding or tissue colonization in infected animals and animal products. We developed and evaluated a quantitative PCR (qPCR) method on spiked sheep tissue and fecal samples for the detection and quantification of Salmonella spp. Without the use of a pre-enrichment step, the qPCR limit of detection (LOD) results for sheep fecal (4 × 104-6 × 103 cfu/g) and tissue (4 × 105-4 × 103 cfu/g) samples were not adequate for detection purposes. With the inclusion of a 6-h pre-enrichment step in buffered peptone water (BPW), the LOD was 9 cfu/g (2.57 × 101 copies/g) in sheep feces, and 5.4 cfu/g (3.22 copies/g) sheep tissue. Comparison of the 6-h BPW qPCR method with a 24-h mannitol-selenite-cystine broth enrichment culture method using spiked samples revealed a sensitivity of 91% and 92%, respectively, and a specificity of 100% for both methods. The correlation was significant between the quantity (copies/mL) of Salmonella spp. in BPW at 6 h and at 0 h, allowing semiquantitative analysis. Our results demonstrate that, following inclusion of a 6-h pre-enrichment step in BPW, qPCR is semiquantitative with improved LODs of Salmonella spp. in sheep fecal and tissue samples.

Erratum: Harris, C.; White, P.J.; Mohler, V.L.; Lomax, S. Electroencephalography Can Distinguish between Pain and Anaesthetic Intervention in Conscious Lambs Undergoing Castration. Animals 2020, 10, 428.

The authors wish to make the following corrections to this paper [...].

Electroencephalography Can Distinguish between Pain and Anaesthetic Intervention in Conscious Lambs Undergoing Castration.

Australian sheep routinely undergo painful surgical husbandry procedures without anaesthesia or analgesia. Electroencephalography (EEG) has been shown to be a successful measure of pain in livestock under a general anaesthetic. The aim of this study was to compare this EEG model to that of conscious lambs undergoing castration with and without local anaesthesia. Sixteen merino crossbred ram lambs 6 to 8 weeks of age (13.81kg ± 1.97) were used in the study. Lambs were randomly allocated to 1 of 4 treatment groups: (1) Conscious EEG and surgical castration with no anaesthetic intervention (CON; n = 4); (2) Conscious EEG and surgical castration with pre-operative applied intra-testicular lignocaine injection (CON + LIG; n = 4); (3) surgical castration under minimal anaesthesia (MAM; n = 4); (4) and surgical castration with pre-operative lignocaine injection (2 mL lignocaine hydrochloride 20 mg/mL, under minimal anaesthesia (MAM + LIG; n = 4). Distinct differences in the EEG parameters Ptot, F50 and F95 between pre-and post-castration in conscious lambs were demonstrated in this study (p < 0.01). Further, CON and CON + LIG treatments were distinguishable using F50 and F95 measures (p = 0.02, p = 0.04, respectively). Significant changes in the EEG output of MAM animals were identified pre- to post-castration (p < 0.01). The EEG output of MAM and MAM + LIG were similar. EEG was successful in differentiating lambs treated with pain relief in a conscious state after castration by examining F50 and F95, which may suggest the suitability of conscious EEG pain measurement.

Salmonella in calves.

Salmonellae are endemic on most large intensive farms and salmonellosis is a common cause of neonatal morbidity and mortality. Disease and mortality usually reflect a variety of management events and environmental stressors that contribute to compromised host immunity and increased pathogen exposure. The diversity of salmonella serovars present on farms, and the potential for different serovars to possess different virulence factors, require the implementation of broad prophylactic strategies that are efficacious for all salmonellae. This article discusses strategies to promote host immunity and minimize pathogen exposure at the farm level. The benefits of control include a reduction in disease incidence and mortality, reduced drug and labor costs, and improved growth rates.

Cross-protective immunity in calves conferred by a DNA adenine methylase deficient Salmonellaenterica serovar Typhimurium vaccine.

The global trend towards intensive livestock production is associated with increased fecal oral pathogen transmission resulting in a high prevalence of Salmonella. Since many pathogenic Salmonella serovars are often endemic to livestock production systems, it is desirable to develop a vaccine that is capable of eliciting immunity to more than one serovar. Here we examined whether immunization with a modified live Salmonella enterica serovar Typhimurium vaccine strain lacking the DNA adenine methylase (Dam) conferred protection in calves against a heterologous S. enterica Dublin challenge. Vaccinated animals challenged with a virulent Dublin strain exhibited a significant attenuation of clinical disease (improved attitude scores and reduced fever and diarrhea) and a concomitant reduction in Dublin fecal shedding and colonization of mesenteric lymph nodes (MLN) compared to non-vaccinated control animals. These data suggest that vaccination with a dam(-) Typhimurium vaccine strain conferred significant cross-protection against clinical disease in cattle attributable to heterologous challenge with Dublin.