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.     

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.     

Technical note: Constrained optimization of breed composition in composite populations to balance net merit and risk

We developed a method to derive optimum composite genotypes when the proportion of a specific breed (or combination of breeds) in the composite is constrained to a desired level. The method allows development of composites that balance net merit according to available parameters and reflects the production system and(or) risk preference, particularly when traits in the breeding objective do not represent all traits important for production. The method can also be used to evaluate the cost of constraining breed use. For example, when the true optimal composite is difficult to obtain in the time frame available to the producer, the predicted merit of the composite with breed proportion restricted to those that are obtainable in practice can be compared with the predicted merit of the true optimum composite. In a case study the method is used to optimize a tropical composite using Brahman, Charolais, and Belmont Red breeds. The proportion of Charolais is to be constrained to 50%. Using growth parameters from a tropical crossbreeding experiment, the unconstrained optimum composite was 32.5% Brahman, 11.1% Belmont Red, and 56.3% Charolais. When the proportion of Charolais in the composite was constrained to 50%, optimum proportions of Brahman and Belmont Red were 33.9% and 16.1% respectively. In the case study net merit (annualized profit per cow exposed, $), as defined by the growth parameters, was .2% less than that of the unconstrained optimum composite.     

Behavioural and physiological responses of sheep to 16 h transport and a novel environment post-transport

The effect of a novel lairage environment on the ability of sheep to recover from 16 h of transport was investigated. Sheep were transported from grass paddocks to either novel outside paddocks or inside pens, and housed groups were transported to either familiar or novel inside pens. During transport, sheep from outside paddocks lay down less than those from inside pens. In sheep transported to inside pens, those from outside paddocks spent more time lying and spent less time eating; hay and water intakes during the first 12 h post-transport were lower than those previously kept inside. There was no obvious effect of a novel environment post-transport on blood biochemistry, suggesting that the lower post-transport feed and water intakes in a novel environment did not have a significant effect on the ability of the sheep to recover from the feed and water deprivation associated with transport.     

Effects of pasteurization of colostrum on subsequent serum lactoferrin concentration and neutrophil superoxide production in calves

OBJECTIVE: To determine the effects of pasteurization of colostrum on serum lactoferrin concentration and neutrophil oxidative function by comparing values from calves given pasteurized (76 C, 15 minutes) colostrum versus calves given fresh frozen colostrum. ANIMALS: 8 Holstein bull calves were used to study the effects of pasteurization of colostrum on the absorption of lactoferrin and neutrophil oxidative burst. Three additional calves were used to study the effect of exogenous lactoferrin on neutrophil oxidative burst. METHODS: Calves were fed fresh frozen or heat pasteurized colostrum (76 C for 15 minutes) via esophageal feeder within 4 hours of birth. Neutrophils were isolated from whole blood samples. Neutrophil oxidative burst was induced by phorbol ester (300 ng/ml) stimulation of cells (1 X 10(6) cells) at 37 C. Serum lactoferrin concentrations were compared, using immunoblot analysis. Serum IgG concentrations were determined by radial immunoassay. Comparisons were made between the use of the 2 types of colostrum in calves by measuring subsequent serum IgG and lactoferrin concentrations and neutrophil superoxide production. RESULTS: Serum IgG and lactoferrin concentrations increased more in calves receiving fresh frozen colostrum. Neutrophil superoxide production was higher in neutrophils prepared from calves receiving fresh frozen colostrum. Colostral lactoferrin addition to neutrophil incubations resulted in increased oxidative burst. CONCLUSIONS AND CLINICAL RELEVANCE: Compared with calves given fresh frozen colostrum, calves given pasteurized colostrum had decreased serum IgG and lactoferrin concentrations and neutrophil superoxide production 24 hours after administration. These results suggest that pasteurizing bovine colostrum at 76 C for 15 minutes has substantial effects on passive transfer of proteins and neutrophil function.     

Effects of hexavalent chromium on development of crabs,Rhithropanopeus harrisii and Callinectes sapidus

Survival of Rhithropanopeus harrisii larvae from hatching to first crab stage occurred in Na₂CrO₄ concentrations from 1.1 to 29.1 ppm. Estimated LC50 for complete zoeal development was 17.8 ppm Na₂CrO₄ and it was 13.7 ppm for development to first crab stage. A concentration of 1.1 ppm Na₂CrO₄ was nontoxic, while Na₂CrO₄ concentrations of 7.2 and 14.5 ppm were sublethal and concentrations of 29.1 to 58.1 ppm were acutely toxic. Low concentrations of Na₂Cr0₄ caused an increase in swimming speed and high concentrations caused a decline. Survival of Callinectes sapidus larvae occurred in Na₂CrO₄ concentrations from 1.1 to 4.7 ppm. The LC50 for complete zoeal development was estimated to be 2.9 ppm Na₂CrO₄ and the LC50 for development to first crab stage was estimated to be 1.0 ppm Na₂CrO₄ The total Cr in sodium chromate is 32% by weight (Tacey,1981), hence, the total Cr concentrations tested were 32% of the Cr salts given above. Statistical analyses of the data on survival, duration and mortality of larvae are presented.     

Mutations in the juxtamembrane domain of c-KIT are associated with higher grade mast cell tumors in dogs

Mast cell tumors are among the most commonly seen tumors of the skin in dogs and are more highly aggressive than mast cell tumors of other species. Some breeds display a markedly higher incidence of mast cell tumor development than others and appear to have some genetic predisposition. Recently, mutations have been found in canine mast cell tumor tissues and cell lines within the juxtamembrane domain of the protooncogene c-KIT In previous studies utilizing a small number of cases, no association between the presence of a mutation and the breed of dog or grade of the tumor could be identified. An expanded study with a larger sample set was performed to explore this possibility. The juxtamembrane domain of c-KIT was amplified using the polymerase chain reaction from genomic DNA preparations of 88 paraffin-embedded mast cell tumors from selected breeds. Mutations, consisting of duplications and deletions, were found in 12 of the tumors. A significant association was found between the presence of a mutation and a higher grade of tumor but not between breed and grade or between breed and the presence of a mutation.     

Carbon kinetics of milk formation in Holstein cows in late lactation

Carbon transfer to milk in Holstein cows in late lactation was measured by introducing changes in the natural stable carbon isotope composition of the feed. Six Holstein cows in mid-lactation were placed on a diet naturally low in 13C (-25.0% vs Pee Dee belemnite [PDB] an international carbon isotope standard), based on alfalfa-barley, and six others were placed on a diet naturally enriched in 13C (-11.5% vs PDB), based on corn. After a 7-wk equilibration period on these diets, three cows were switched from alfalfa-barley to corn, and three were switched from corn to alfalfa-barley. The three other cows in each group served as controls. 13C/12C ratios were measured in daily morning milk samples during the week before and for 6 wk after the changes in diet. After the diets had been switched, milk isotope ratios rapidly approached the isotopic composition of the new diet, indicating rapid transfer of dietary carbon into milk. The data were consistent with a model whereby milk was synthesized from a single precursor pool that responded rapidly to dietary perturbation. The milk precursor pool had a half-life of approximately .9 d and had a mass of approximately 7 kg of carbon, which was renewed daily by the entry of 5 kg of digestible dietary carbon.