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.     

Inheritance of virulence in Fusarium circinatum,the cause of pitch canker in pines

Wildtype strains of Fusarium circinatum, the causal agent of pitch canker, were crossed to obtain an F₁ generation. Progeny of this cross were tested for virulence by inoculating Pinus radiata seedlings, and were found to induce a wide range of lesion lengths. Two strains from the F₁ generation that induced long lesions (= high virulence) were used as parents to produce an F₂ generation, followed by a second round of selection for high virulence to obtain an F₃ generation. Mean lesion lengths were not significantly different between the three generations (P ≥ 0.196). A parallel set of crosses was performed to select for low virulence by using progeny in the F₁ and F₂ generations that induced short lesions as parents for F₂ and F₃ generations, respectively. In this case, both rounds of selection resulted in a significant reduction in mean lesion length, from 33.8 ± 0.8 mm in the F₁ generation, to 19.7 ± 0.7 and 12.9 ± 0.7 mm in the F₂ and F₃ generations, respectively. Thus it is apparent that F. circinatum retains the genetic capacity for avirulence to pines, which could reflect a lack of strong selection for virulence in nature. Progeny of a cross between high and low virulence parents manifested nearly continuous variation in lesion lengths, consistent with virulence being a quantitatively inherited trait. Based on this cross, broad‐sense heritability (H²) was determined to be 0.74, which suggests that virulence is under strong genetic control.     

Measurement of glycosaminoglycans and keratan sulphate in canine arthropathies

Glycosaminoglycans (gag) and keratan sulphate (ks) were measured in sera and synovial fluids from dogs with either osteoarthritis (oa) or rupture of the cranial cruciate ligament (ccl) and normal dogs. The dogs with oa had higher synovial fluid gag levels (P<0·002) and serum KS (P<0·03) compared to the normal dogs. No significant differences in serum gag were found in either group. In both oa and rupture of the ccl, gag levels were increased in the synovial fluid from the affected joint compared with the clinically normal (inactive) contralateral joint. Neither gag nor ks measurements correlated with serum and synovial fluid antibodies to collagen type II, synovial fluid white cell count or age of dog. It is unlikely that the measurement of these cartilage breakdown products is of value for diagnostic or prognostic use in canine arthropathies.     

Urethral conformation and position in relation to urinary incontinence in the bitch.

The conformation of the urethra, the position of the bladder neck and the distance between the external urethral orifice and the cranial pubic brim were compared in 30 continent and 30 incontinent bitches with incompetence of the urethral sphincter mechanism, using the measurements made from vagino-urethrograms. The bladder neck was significantly (P less than 0.001) further caudal in incontinent dogs than in continent dogs. Its position was not affected by neutering and could not be explained by the degree of urethral curvature. The distance from the external urethral orifice to the cranial pubic brim was correlated (P less than 0.001) with bodyweight but was not significantly different in the continent and incontinent bitches. In neutered animals, the distance between the cranial pubic brim and the external urethral orifice was significantly (P less than 0.05) shorter than in entire animals.     

Assessment of the ultrasonographic characteristics of the podotrochlear apparatus in clinically normal horses and horses with navicular syndrome

OBJECTIVE: To characterize the normal ultrasonographic appearance of the podotrochlear apparatus in horses by use of standardized measurements and identify soft tissue changes associated with navicular syndrome. DESIGN: Prospective study. ANIMALS: 7 clinically normal horses and 28 horses with navicular syndrome. PROCEDURE: The feasibility of identifying and measuring the soft tissue structures of the podotrochlear apparatus ultrasonographically via the transcuneal approach was assessed in 2 additional horses without navicular syndrome; both horses were euthanatized, and the structures identified ultrasonographically were confirmed at necropsy. Ultrasonographs were obtained in the study horses. Objective and subjective data were obtained to characterize ultrasonographic changes associated with navicular syndrome. RESULTS: Abnormalities of the flexor surface of the distal sesamoid (navicular) bone, the impar ligament, the distal digital annular ligament, deep digital flexor tendon (DDFT), and the podotrochlear (navicular) bursa were assessed via the transcuneal ultrasonographic approach. No significant differences were found between the measurements of the podotrochlear apparatus in normal horses and those with navicular syndrome; however, important subjective differences were detected ultrasonographically in horses with navicular syndrome. In horses with navicular syndrome, ultrasonographic findings were indicative of navicular bursitis, dystrophic mineralization of the DDFT and impar ligament, tendonitis and insertional tenopathy of the DDFT, desmitis of the impar ligament, and cortical changes in the flexor surface of the navicular bone. CONCLUSIONS AND CLINICAL RELEVANCE: Findings of ultrasonographic evaluation of the hoof appear to be useful in determining the cause of caudal heel pain and characterizing the components of navicular syndrome in horses.     

Temporal patterns of growth hormone in blood and seminal plasma of mature dairy bulls

This study was conducted to determine if growth hormone (GH) concentration in bovine seminal plasma would be proportional to but less variable than blood plasma GH. The relationship between GH in blood and seminal plasma was also examined critically. Blood samples were collected at 15-min intervals for 5.75 h, while semen was collected at 30-min intervals over the same time period. Average seminal plasma GH concentrations were 3.2 times higher (P less than .05) than blood plasma GH concentrations (40.4 +/- 15.8 ng/ml vs 12.6 +/- 1.2 ng/ml, respectively). The within animal correlation between blood and seminal plasma was consistently low and nonsignificant (P greater than .05). Overall blood plasma GH and seminal plasma GH concentrations were weakly correlated (r = .418; P greater than .05) among bulls. A predictable relationship between blood and seminal plasma GH concentration does not exist under the conditions of this study.     

A linear functional relationship model for circular data with an application to the assessment of ocean wave measurements

This article suggests a linear functional relationship model for comparing two sets of circular data subject to unobservable errors. Unlike the corresponding and relatively well-studied model for linear data, maximum likelihood estimation for this model is very complicated and no explicit solutions are possible. Using a numerical approximation, we are able to solve the likelihood equations approximately, and to obtain good approximations to the likelihood estimates of the parameters. The quality of our estimates and the feasibility of the estimation method are illustrated via simulation. By establishing a parallel with the model for linear data, we are able to explain the various problems occurring in the process of estimation and to substantiate our numerical results. The interest in the model arose in connection with the study of ocean wave data; an application to such data is also given.