Second round of an interlaboratory comparison of SARS-CoV2 molecular detection assays used by 45 veterinary diagnostic laboratories in the United States

The COVID-19 pandemic presents a continued public health challenge. Veterinary diagnostic laboratories in the United States use RT-rtPCR for animal testing, and many laboratories are certified for testing human samples; hence, ensuring that laboratories have sensitive and specific SARS-CoV2 testing methods is a critical component of the pandemic response. In 2020, the FDA Veterinary Laboratory Investigation and Response Network (Vet-LIRN) led an interlaboratory comparison (ILC1) to help laboratories evaluate their existing RT-rtPCR methods for detecting SARS-CoV2. All participating laboratories were able to detect the viral RNA spiked in buffer and PrimeStore molecular transport medium (MTM). With ILC2, Vet-LIRN extended ILC1 by evaluating analytical sensitivity and specificity of the methods used by participating laboratories to detect 3 SARS-CoV2 variants (B.1; B.1.1.7 [Alpha]; B.1.351 [Beta]) at various copy levels. We analyzed 57 sets of results from 45 laboratories qualitatively and quantitatively according to the principles of ISO 16140-2:2016. More than 95% of analysts detected the SARS-CoV2 RNA in MTM at ≥500 copies for all 3 variants. In addition, for nucleocapsid markers N1 and N2, 81% and 92% of the analysts detected ≤20 copies in the assays, respectively. The analytical specificity of the evaluated methods was >99%. Participating laboratories were able to assess their current method performance, identify possible limitations, and recognize method strengths as part of a continuous learning environment to support the critical need for the reliable diagnosis of COVID-19 in potentially infected animals and humans.     

Presettlement and modern disturbance regimes in coast redwood forests: Implications for the conservation of old-growth stands

Coast redwood (Sequoia sempervirens), a western North American conifer of ancient lineage, has a paradoxical combination of late-successional characteristics and strong adaptations to disturbance. Despite its shade tolerance and heavy dominance of the canopy on many sites, redwood saplings are uncommon in upland old-growth stands. Information needed to ensure the conservation of old-growth redwood forests has been limited. In this review paper, we integrate evidence on redwood biology with data on the historic and modern disturbance regimes to help clarify the degree to which key attributes of redwood forests may have been dependent upon periodic disturbance. Available evidence suggests that episodes of fire, flooding, and slope failure prior to European settlement were frequent but predominantly of low to moderate severity and extent, resulting in broadly uneven-aged forests. The majority of fires prior to European settlement were apparently of human origin. Frequency and severity of the major disturbance agents have been radically changed in modern times. Fires have been largely excluded, and flooding has been altered in ways that have often been detrimental to old-growth redwoods on alluvial terraces. However, because of the apparent anthropogenic origin of most presettlement fires, the long-term evolutionary role of fire for coast redwood is ecologically ambiguous. With fire exclusion, redwood possibly could be displaced to some extent on upland sites by increasing abundance of fire-sensitive competitors. Alternatively, redwood may be able to maintain dominance by vegetative sprouting and new seedling establishment on root-wad mounds, fallen logs, and on soil exposed by slope failure. Future research priorities are suggested that will help resolve some of the current ambiguities.     

Feline one-way mixed leukocyte reaction

The feline one-way mixed leukocyte reaction (MLR) was accomplished by a microtechnique assay using gradient purified mononuclear leukocytes from the peripheral blood of specific-pathogen-free (SPF) cats. An eight day assay was required with an appropriate serum supplement of rabbit or cat serum and obtainable cell concentrations of 1×10⁵ mononuclear cells per well of each responder and stimulator population. This assay was devised as an $\text{in vitro}$ correlate of cell-mediated immunity and to demonstrate that feline histocompatibility differences can be detected in culture.     

Prevalence of Giardia spp. infection in pre-weaned and weaned calves in relation to management factors

Two hundred and forty calf faecal samples from 16 Malaysian farms were screened by PCR for Giardia spp. The overall prevalence was 12.5% and the overall farm prevalence was 68.8% (11/16 farms). The prevalence in pre-weaned and weaned calves was 16.7% and 8.3%, respectively. Sequence analysis of 25 isolates identified all as G. duodenalis assemblage E. Management factors associated with an increased risk of infection with Giardia spp. included keeping weaned calves in pens with sand floors and calf age. Keeping pre-weaned calves in pens with concrete floors and calving in single cow calving areas decreased the risk.     

Spontaneous gastroduodenal perforation in 16 dogs and seven cats (1982-1999)

The records of 23 dogs and cats diagnosed with spontaneous gastroduodenal perforation (GDP) were retrospectively reviewed. Survival was 63% in dogs and 14% in cats. Rottweilers <5 years of age were overrepresented. Clinical evidence of gastrointestinal bleeding was common in dogs but not in cats. Shock was an uncommon presenting condition in dogs and was not closely linked to outcome. In fact, progression of an ulcerating lesion to GDP was not associated with marked changes in symptoms exhibited by many patients in this study. Most GDPs were associated with histopathological evidence of subacute or chronic peritoneal reaction at the time of diagnosis. This suggests that diagnostic methods employed lacked sensitivity in identifying early perforating lesions, and that dramatic signs of acute abdomen following gastroduodenal perforation may not be as common as was previously thought.     

Heinz body anemia in a dog that had been sprayed with skunk musk

A 5-year-old pit bull-type dog was examined because of lethargy and black feces, and a presumptive diagnosis of methemoglobinemia was made on the basis of grossly visible brown discoloration of the mucous membranes and urine. Heinz body formation was seen on examination of blood smears, and severe anemia developed 3 days later. The dog did not have any history of exposure to toxins or drugs that have been associated with Heinz body formation. The only remarkable historical finding was that the dog had been wandering loose the day prior to initial examination and returned home smelling strongly of skunk spray. Skunk spray contains thiols and other compounds that are strong oxidizing agents. It was speculated that the methemoglobinemia and Heinz body anemia were a result of exposure to these compounds. Skunk interactions with dogs usually have malodorous but otherwise harmless results. However, findings in this dog suggest that more serious consequences may develop in isolated cases.     

Evaluation of ilial screw loosening after triple pelvic osteotomy in dogs: 227 cases (1991-1999)

OBJECTIVE: To investigate factors influencing screw loosening after triple pelvic osteotomy (TPO) and ischial wire stabilization of the acetabular segment. DESIGN: Retrospective case series. Animals-227 dogs with congenital hip dysplasia or subluxated hip joints. PROCEDURES: Medical records and radiographs of 227 dogs that underwent 332 TPO procedures were evaluated, and data pertaining to screw type, plate position, sacral screw engagement, use of ischial interfragmentary wires, and pelvic alignment were assessed for associations with screw loosening. RESULTS: Complications developed in 96 of the 332 (29%) procedures. Cancellous screws without sacral engagement were associated with the lowest frequency (6%) of loosening, compared with cancellous and cortical screws engaging the sacrum and cortical screws that did not engage the sacrum. Frequency of screw loosening increased when cortical or cancellous screws engaged the sacrum and when cortical screws were used. In dogs that had surgery bilaterally, the first limb on which TPO was performed had a higher frequency of screw loosening than the second limb. Pelvic alignment loss was greatest (5.4 degrees ) when the 3 most cranial screws were loosened. Loss of pelvic alignment was significantly different between dogs that underwent surgery and had complications and those that underwent surgery and did not have complications in association with loosening of 1, 2, and 3 screws. CONCLUSIONS AND CLINICAL RELEVANCE: TPO screw loosening was multifactorial and related to stability of the affected ilium, screw type, and screw position. Placing cancellous screws that do not engage the sacrum in pelvic osteotomy plate positions 1 through 3 may decrease the number of screws that loosen.     

Urodynamic effects of a percutaneously controlled static hydraulic urethral sphincter in canine cadavers

OBJECTIVE: To describe a percutaneously controlled static hydraulic urethral sphincter (SHUS) and evaluate urodynamic effects of the SHUS in canine cadavers. SAMPLE POPULATION: Cadavers of 6 adult female dogs. PROCEDURE: Cadavers were obtained immediately after dogs were euthanatized. Baseline maximal urethral closure pressure (MUCP) and cystourethral leak point pressure (CLPP) were measured by use of a urethral pressure profilometer. An SHUS system was constructed by use of a silicone vascular occluder and subcutaneous infusion port. The SHUS system was then placed around the pelvic urethra in each cadaver. Measurements of MUCP and CLPP were repeated after varying occlusion of the SHUS (0%, 25%, and 50% occlusion). Baseline MUCP and CLPP values were compared with values obtained at 0%, 25%, and 50% occlusion of the SHUS by use of repeated-measures ANOVA. RESULTS: Mean +/- SD MUCP for canine cadavers was 7 +/- 1.3 cm H2O at baseline, which increased to 127 +/- 53 cm H2O after 50% occlusion of the SHUS. Mean CLPP was 11 +/- 8.6 cm H2O at baseline, which increased to 73 +/- 38 cm H2O after 50% occlusion of the SHUS. Mean MUCP and CLPP were significantly associated with the amount of occlusion. CONCLUSIONS AND CLINICAL RELEVANCE: The SHUS had positive effects on MUCP and CLPP in canine cadavers. Therefore, additional evaluation of the SHUS in live dogs is warranted.