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.     

Liver haemorrhages induced by rapeseed meal: incidence in adult male and female fowls

Adult cockerels of a commercial egg-producing strain did not develop liver haemorrhages when maintained for 12 weeks on a diet containing 400 g rapeseed meal/kg food. Haemorrhages occurred in 43% of laying hens of the same strain as the cockerels and 33% of laying hens of another strain maintained on the same diet for the same period. Oestrogenisation of the male birds did not influence the incidence of haemorrhage.     

Effect of Fusarium roseum corn culture containing zearalenone on early pregnancy in swine

A corn culture of Fusarium roseum was added to a standard corn-soybean swine gestation ration. Low, middle, and high dosage mixed feeds contained 7, 38, and 64 mg of zearalenone/kg of feed (7, 38, and 64 ppm) and 0.5, 2.5, and 4.5 mg of deoxynivalenol/kg, respectively. Control feed was the standard ration without added F roseum corn culture. Mature gilts were bred by natural service and fed control or F roseum molded feed from 3 to 34 days after breeding. The main effect of the molded feed was an inhibition of fetal development, with decreased numbers of fetuses present in treated animals at slaughter (38 to 43 days after breeding). Normal litters were present in 7 of 8 control animals, in 2 of 4 gilts given the low-dosage feed, in 1 of 4 gilts given the medium dosage, and in 0 of 4 given the high-dosage feed. Corpora lutea were maintained in all treated animals, as evidenced by serum progesterone concentrations. Serum estradiol concentrations were decreased in gilts in the middle- and high-dosage groups. The genital system of the gilts fed low- and middle-dosage feeds had a gross and microscopic appearance similar to that of the pregnant controls and reflected prolonged progesterone stimulation. Morphologic changes in the genital system of the high-dosage group were intermediate between changes induced by progesterone and those induced by estrogen. Clinical signs of hyperestrogenism and partial feed refusal were noticed in only some of the high-dosage group animals.     

Comparison of three serologic techniques for detection of antibody to Mycoplasma pulmonis

A comparison of three micro-assay detection systems for naturally occurring and experimentally induced rat antibody to Mycoplasma pulmonis was performed. The micro-enzyme linked immunoabsorbent assay was found superior to counter immunoelectrophoresis or complement fixation techniques when comparing sensitivity, duration of antibody response, or technical requirements. The primary immune response was routinely detected at 3 weeks after antigen exposure when using the enzyme linked or complement fixation assays.     

Recurrence of thoracolumbar intervertebral disk extrusion in chondrodystrophic dogs after surgical decompression with or without prophylactic fenestration: 265 cases (1995-1999)

OBJECTIVES: To evaluate a population of chondrodystrophic dogs treated for Hansen type 1 intervertebral disk (IVD) disease by surgical decompression with or without prophylactic fenestration and determine the rate and location of surgically confirmed recurrence of intervertebral disk extrusion. ANIMALS: 265 dogs. STUDY DESIGN: Retrospective study. PROCEDURES: Medical records of dogs that underwent spinal decompression between 1995 and 1999 were reviewed. RESULTS: 3 (4.9%) dogs were euthanatized or died prior to discharge. Fenestration was performed in 252 dogs, including 37 (14.7%) at the site of decompression only, 48 (19%) at 3 to 4 disk spaces, and 167 (66%) at 5 to 7 disk spaces. There were 12 instances of recurrent disk extrusion confirmed by removal of disk at a second surgery 3.5 to 33 months after the first surgery. Recurrence was always at a new disk space, and rates did not significantly differ between dogs that underwent single or multiple fenestrations. Two recurrences were at a previously fenestrated disk space. Seven recurrences were at a site immediately adjacent to a fenestrated disk space, and 5 recurrences were at L4-5. CONCLUSIONS AND CLINICAL RELEVANCE: Prophylactic fenestration is generally successful in preventing future disk extrusions at fenestrated disk spaces. Prospective evaluation is still required to determine whether fenestration decreases the overall rate of recurrence. Prophylactic fenestration could promote disk extrusion at adjacent, nonfenestrated disk spaces. This could have a substantial clinical impact if recurrence develops at L4-5.     

Effect of Fusarium roseum (Gibberella zea) on pregnancy and the estrous cycle in gilts fed molded corn on days 7-17 post-estrus

A laboratory produced corn culture of Fusarium roseum (Gibberella zea) was fed as 10% of the total diet to bred and non-bred gilts on days 7-17 postestrus. The corn culture contained both zearalenone and deoxynivalenol by analysis, and caused considerable feed refusal. Although 6 of 10 bred gilts fed the corn culture appeared to be pregnant by ultrasound testing between days 40-50 post-breeding, none of them farrowed. Progesterone analyses indicated that none of the 10 gilts cycled normally during the 20 week observation period. Three maintained high serum progesterone (greater than 10 ng/ml) during the entire period. Progesterone analyses were similar in 5 non-bred gilts fed the corn culture in that none of them cycled normally in 20 weeks. The retention or loss of corpora lutea could not be attributed to levels of luteinizing hormone, as no relationship was found between concentrations of progesterone and luteinizing hormone. This experiment indicates that limited dietary exposure of female swine to Fusarium roseum infected corn can result in embryonic loss and disruption of normal reproductive cycling for an extended length of time.     

Septic peritonitis and uroperitoneum secondary to subclinical omphalitis and concurrent necrotizing cystitis in a colt

A 15-day-old American Quarter horse colt was presented for depression and pyrexia. Peritonitis was diagnosed following peritoneal fluid analysis. Exploratory laparotomy revealed an area of focal necrosis over the dorsal wall of the urinary bladder leading to peritonitis and uroperitoneum. The affected area of the urinary bladder was resected and the peritonitis resolved with medical treatment.     

Impact of diet on development of bronchial-associated immunity in the neonatal piglet

Bronchial-associated immune development is critically important to protect neonates from respiratory infections. Herein, bronchial-associated immune development in formula-fed and sow-reared pigs is described. Colostrum-fed newborn piglets were fed medicated sow milk replacer formula beginning at 48 h of life or remained with the sow. Blood and tissues were sampled at one-week (d7) and three-weeks (d21) of age. Lymphocyte subpopulations, including T helper 2, cytotoxic T, memory T, and NK cells, in peripheral blood, mediastinal lymph nodes, and thoracic lymph nodes were identified using flow cytometry. Additionally, IL-1β, IL-6, IL-12, TNFα, TGF-β1, TGF-β2, IFNα, IFNβ, and dectin gene expression were analyzed by quantitative RT-PCR. Total IgG, IgM, and IgA concentrations in serum were analyzed. Dietary and developmental effects were observed. This set of baseline measurements provides a framework for future respiratory challenge studies where the effects of diet on the neonate's ability to resist and/or recover from infection can be tested.     

Effect of adrenalectomy on cholecystokinin-8-induced Fos-like immunoreactivity in myenteric neurons and the dorsal vagal complex in rats

OBJECTIVE: To investigate the effect of adrenalectomy on cholecystokinin-8 (CCK-8)-induced Fos-like immunoreactivity (Fos-LI) in the myenteric neurons of the dorsal vagal complex (DVC) in rats. ANIMALS: 16 male Sprague Dawley rats. PROCEDURES: Rats were allocated to 1 of 2 groups and underwent adrenalectomy or a sham adrenalectomy procedure. Rats were challenged with a supraphysiologic dose of CCK-8 (40 microg/kg) or physiologic saline (0.9% NaCl) solution (0.5 mL) administered IP; after 90 minutes, rats were euthanized, and Fos-LI was quantified in the DVC (at the levels of the area postrema, nucleus tractus solitarii, and dorsal motor nucleus of the vagus) and the myenteric neurons of the duodenum and jejunum by use of a diaminobenzidine reaction enhanced with nickel. The Fos-LI-positive cells were counted by use of an automated system and manually in the DVC and intestinal samples, respectively. Counts of Fos-LI in the different hindbrain levels and myenteric neurons were compared between the adrenalectomy--and shamtreated groups and between the CCK-8- and saline solution-treated groups. RESULTS: After adrenalectomy, CCK-8-induced Fos-LI was attenuated only in the myenteric neurons of the duodenum. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that the adrenal gland has a role in the activation of myenteric neurons by CCK-8 in rats.     

Morphologic and biochemical characterization of hyperextension of the metacarpophalangeal and metatarsophalangeal joints in llamas

OBJECTIVE: To determine the morphologic and biochemical characteristics of hyperextension of the metacarpophalangeal and metatarsophalangeal joints in llamas. ANIMALS: 12 adult llamas (6 with bilateral hyperextension of the metacarpophalangeal or metatarsophalangeal joints and 6 age- and sex-matched control llamas). PROCEDURES: Llamas were evaluated by use of lameness examination, ultrasonography, and radiography. A CBC, serum biochemical analysis, and determination of concentrations of trace minerals in serum and liver samples were performed. Llamas were euthanized, and samples of the superficial digital flexor tendon, deep digital flexor tendon, and suspensory ligament were obtained from 4 areas and snap-frozen in liquid nitrogen or suspended in neutral-buffered 10% formalin. Immunohistochemical evaluation of collagen types I and III and assays for measurement of lysyl oxidase activity were performed. RESULTS: 2 affected llamas had a visible gait deficit associated with metacarpophalangeal joint hyperextension. Radiographic evidence of osteoarthritis was detected in 1 severely affected llama, and ultrasonographic changes of soft tissue mineralization and suspensory desmitis were observed in 2 llamas. Liver concentrations of copper were lower and serum concentrations of zinc higher in affected llamas, compared with values in control llamas. Lysyl oxidase activity and collagen distribution did not differ significantly between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Hyperextension of the metacarpophalangeal or metatarsophalangeal joints in llamas does not appear to be the result of injury or degeneration of the suspensory ligament or flexor tendons. Lower copper concentrations coupled with higher zinc concentrations in affected llamas may be indicative of secondary copper deficiency.