How viral and host factors contribute to the severe pathogenicity of the H5N1 subtype of avian influenza virus infection in humans is poorly understood. We identified three clusters of differentially expressed innate immune response genes in lungs from H5N1 (A/Vietnam/1203/04) influenza virus-infected ferrets by oligonucleotide microarray analysis. Interferon response genes were more strongly expressed in H5N1-infected ferret lungs than in lungs from ferrets infected with the less pathogenic H3N2 subtype. In particular, robust CXCL10 gene expression in H5N1-infected ferrets led us to test the pathogenic role of signaling via CXCL10's cognate receptor, CXCR3, during H5N1 influenza virus infection. Treatment of H5N1-infected ferrets with the drug AMG487, a CXCR3 antagonist, resulted in a reduction of symptom severity and delayed mortality compared to vehicle treatment. We contend that unregulated host interferon responses are at least partially responsible for the severity of H5N1 infection and provide evidence that attenuating the CXCR3 signaling pathway improves the clinical course of H5N1 infection in ferrets.

Farber disease (FD) is a severe inherited disorder of lipid metabolism characterized by deficient lysosomal acid ceramidase (ACDase) activity, resulting in ceramide accumulation. Ceramide and metabolites have roles in cell apoptosis and proliferation. We introduced a single-nucleotide mutation identified in human FD patients into the murine Asah1 gene to generate the first model of systemic ACDase deficiency. Homozygous Asah1P361R/P361R animals showed ACDase defects, accumulated ceramide, demonstrated FD manifestations and died within 7–13 weeks. Mechanistically, MCP-1 levels were increased and tissues were replete with lipid-laden macrophages. Treatment of neonates with a single injection of human ACDase-encoding lentivector diminished the severity of the disease as highlighted by enhanced growth, decreased ceramide, lessened cellular infiltrations and increased lifespans. This model of ACDase deficiency offers insights into the pathophysiology of FD and the roles of ACDase, ceramide and related sphingolipids in cell signaling and growth, as well as facilitates the development of therapy.

Facial expressions are increasingly being used to assess pain in non-human species, including rodents, horses, and lambs. The development of these species-specific grimace scales has allowed for more rapid pain detection, which can lead to better animal welfare if intervention promptly occurs. For grimace scales to ever be used as a stand-alone measure of pain, it is important they correlate with established pain assessment tools, such as behavioral analysis. This preliminary study aimed to determine whether piglets exhibit pain grimacing and if these facial expressions correlate with their behavior. It also assessed and compared the behavior of boar piglets given an analgesic and topical anesthetic prior to surgical castration and tail docking to piglets that did not receive anything for pain relief. Five-day-old male Yorkshire piglets (n = 19) from four pens were randomly assigned, within their pen, to one of five possible treatments: meloxicam (0.4 mg/kg, intramuscularly) + EMLA® cream, meloxicam (0.4 mg/kg, intramuscularly) + non-medicated cream, saline (intramuscularly) + EMLA® cream, saline (intramuscularly) + non-medicated cream, or no treatment prior to surgical castration and tail docking. Piglet behaviors were video recorded for 8 h immediately after castration, as well as for 1 h at 24 h pre- and post-castration. Their individual behaviors were scored continuously for the first 15 min of every hour of video collected. Facial images were also captured across all time points. A Piglet Grimace Scale (PGS) was developed and used by two observers blinded to treatment, time, and procedure to score over 600 piglet faces. All piglets displayed significant behavioral changes up to 7 h post-castration when compared to baseline, and the use of meloxicam and EMLA® cream was not associated with a reduction in painful behaviors. Significantly higher PGS scores were noted at 0, 3, 4, and 5 h post-castration when compared to PGS scores at 7 h and there was no effect of treatment. PGS scores significantly correlated with piglet behavioral activity. The results suggest that the PGS may have utility for pain evaluation in neonatal pigs.

No abstract

Ensuring that laboratory rodent pain is well managed underpins the ethical acceptability of working with these animals in research. Appropriate treatment of pain in laboratory rodents requires accurate assessments of the presence or absence of pain to the extent possible. This can be challenging some situations because laboratory rodents are prey species that may show subtle signs of pain. Although a number of standard algesiometry assays have been used to assess evoked pain responses in rodents for many decades, these methods likely represent an oversimplification of pain assessment and many require animal handling during testing, which can result in stress-induced analgesia. More recent pain assessment methods, such as the use of ethograms, facial grimace scoring, burrowing, and nest-building, focus on evaluating changes in spontaneous behaviors or activities of rodents in their home environments. Many of these assessment methods are time-consuming to conduct. While many of these newer tests show promise for providing a more accurate assessment of pain, most require more study to determine their reliability and sensitivity across a broad range of experimental conditions, as well as between species and strains of animals. Regular observation of laboratory rodents before and after painful procedures with consistent use of 2 or more assessment methods is likely to improve pain detection and lead to improved treatment and care—a primary goal for improving overall animal welfare.