Suggested guidelines for immunohistochemical techniques in veterinary diagnostic laboratories

This document is the consensus of the American Association of Veterinary Laboratory Diagnosticians (AAVLD) Subcommittee on Standardization of Immunohistochemistry on a set of guidelines for immunohistochemistry (IHC) testing in veterinary laboratories. Immunohistochemistry is a powerful ancillary methodology frequently used in many veterinary laboratories for both diagnostic and research purposes. However, neither standardization nor validation of IHC tests has been completely achieved in veterinary medicine. This document addresses both issues. Topics covered include antibody selection, fixation, antigen retrieval, antibody incubation, antibody dilutions, tissue and reagent controls, buffers, and detection systems. The validation of an IHC test is addressed for both infectious diseases and neoplastic processes. In addition, storage and handling of IHC reagents, interpretation, quality control and assurance, and troubleshooting are also discussed. Proper standardization and validation of IHC will improve the quality of diagnostics in veterinary laboratories.     

Gas exchange,water relations,and Ion concentrations of salt‐stressed tomato and melon plants

Abstract

Tomato and melon plants were grown in a greenhouse and irrigated with nutrient solution having an EC of 2 dS m^?1 (control treatment) and 4, 6, and 8 dS m^?1, produced by adding NaCl to the control nutrient solution. After 84 days, leaf water relations, gas exchange parameters, and ion concentrations, as well as plant growth, were measured. Melon plants showed a greater reduction in shoot weight and leaf area than tomato at the two highest salinity levels used (6 and 8 dS m^?1). Net photosynthesis (Pn) in melon plants tended to be lower than in tomato, for all saline treatments tested. Pn was reduced by 32% in melon plants grown in nutrient solution having an EC of 4 dS m^?1, relative to control plants, and no further decline occurred at higher EC levels. In tomato plants, the Pn decline occurred at EC of 6 dS m^?1, and no further reduction was detected at EC of 8 dS m^?1. The significant reductions in Pn corresponded to similar leaf Cl^? concentrations (around 409 mmol kg^?1 dry weight) in both plant species. Net Pn and stomatal conductance were linearly correlated in both tomato and melon plants, Pn being more sensitive to changes in stomatal conductance (g_s) in melon than in tomato leaves. The decline in the growth parameters caused by salinity in melon and tomato plants was influenced by other factors in addition to reduction in Pn rates. Melon leaves accumulated larger amounts of Cl^? than tomato, which caused a greater reduction in growth and a reduction in Pn at lower salinity levels than in tomato plants. These facts indicate that tomato is more salt‐tolerant than melon.     

Profitability of fungicide applications for managing soybean rust in scenarios of variable efficacy and costs: A stochastic simulation

Soybean rust (SBR) in Brazil is controlled with fungicides, which have shown variable, sometimes declining, efficacy over time. A Monte Carlo simulation framework was implemented as a decision tool and to estimate the probability for a fungicide programme being profitable depending on efficacy and total cost defined by the user. Probability distributions were fitted to slopes and intercepts of the disease–yield relationship and severity in the untreated plots reported in the literature, as well as historical records of soybean price. Simulations of disease reduction conditioned to predefined control efficacy and total application costs were split into scenarios that combined two categories of severity (high and low) and two attainable yield classes (high and low). These categories were defined based on the median of severity (57.8%) and median of the intercept (yield when severity is zero, 2995.1 kg/ha). Probability matrices were constructed relating fungicide efficacy and costs. A higher frequency of break-even events occurred in scenarios of high disease pressure and higher yield. Yearly simulations, starting with 79.4% efficacy, assuming two rates of decline previously determined for tebuconazole (high decline rate), showed that the programme may remain profitable during the first 5 to 7 years of use, in contrast to cyproconazole (low decline rate), a fungicide that would be profitable during the entire decade. This approach was shown to be useful and can be adapted to other diseases of soybean and other crops, as long as damage functions are available. An interactive web app was developed to perform the simulations accessible at alvesks.shinyapps.io/rusty-profits/.     

Cutaneous neosporosis in two adult dogs on chronic immunosuppressive therapy.

Antemortem diagnosis of generalized ulcerative and pyogranulomatous dermatitis with numerous intralesional tachyzoites was made from skin biopsy specimens from 2 adult dogs on chronic immunosuppressive therapy. A 9-year-old Italian Greyhound was on long-term corticosteroid therapy for the treatment of a lupus-like systemic autoimmune disorder, and a 7-year-old Labrador Retriever had received several months of chemotherapy for lymphosarcoma. The tachyzoites were identified as Neospora caninum by immunoperoxidase immunohistochemistry. Both dogs were treated with clindamycin. Lesions in the Greyhound resolved; however, the Labrador Retriever was euthanized because of evidence of neuromuscular disease, despite improvement of the skin lesions. These 2 cases indicate that cutaneous neosporosis can occur in adult dogs on chronic immunosuppressive therapy. The disease may result from reactivation of a congenital infection and/or a recently acquired primary infection.     

Growth and yield modelling of <Emphasis Type="Italic">Acacia mangium</Emphasis> in Colombia

To study the growth and yield of Acacia mangium in the Caribbean region of Colombia, allometric equations of total volume and aboveground plus coarse roots biomass were fitted as a function of the tree’s diameter at breast height (dbh). The von Bertalanffy’s growth model and 59, 0.1 ha plots (0.55–9.55 years old) were used to develop site index (SI) curves at 6 years base age. Then, using the state-space approach, stand growth and yield models were developed for basal area, volume and biomass. The results show that A. mangium is a very promising species for timber production, atmospheric carbon removal and soil restoration because it grows very fast even in mining degraded soils. On average sites it reaches 15 m in height in 3 years. However, early and reiterated thinning coupled with initial mortality by cattle invasion of the very young understocked plantations are producing relatively low yields.     

Eastern equine encephalitis in 9 South American camelids

BACKGROUND: Eastern equine encephalitis (EEE) virus is a mosquito-borne togavirus (alphavirus) that causes severe (often fatal) encephalitis in many mammalian species, but it has not been reported previously in South American camelids. Hypothesis: South American camelids can become naturally infected with EEE virus and show encephalitic signs similar to those observed in other affected species. ANIMALS: Nine cases (8 alpacas and 1 llama, aged 3.5 weeks to 12 years) were identified; 4 of 9 were 510 weeks old. All cases were from the East Coast of the United States and presented in late summer and fall. METHODS: A retrospective study was performed to include confirmed cases of EEE in camelids in North America before 2006. RESULTS: Eight of nine (89%) camelids died or were euthanized in extremis, with the mean time to death of 2 days. Clinical signs were consistent with encephalitis and included fever, lethargy, ataxia, seizures, recumbency, torticollis, opisthotonus, and vestibular signs. No consistent hematologic abnormalities were identified, and cerebrospinal fluid contained an increased protein concentration in the single camelid analyzed. No successful therapy was identified. EEE was confirmed by alphavirus detection by using immunohistochemistry (IHC) and polymerase chain reaction (PCR) in the central nervous system (CNS) and by serology. Findings included polioencephalitis with lymphocytic perivascular cuffing; neutrophil infiltration; gliosis; neuron satellitosis; necrosis; and edema, with intracytoplasmic alphavirus within neurons and glial cells. No virus was detected in extraneural tissues. CONCLUSIONS AND CLINICAL IMPORTANCE: In endemic areas, EEE should be considered a differential diagnosis for young and adult camelids with CNS disease. Brain histopathology with indirect IHC or PCR is diagnostic.     

Impact of seed blend and structured maize refuge on Helicoverpa zea (Lepidoptera: Noctuidae) potential phenological resistance development parameters in pupae and adults

BACKGROUND

Helicoverpa zea, an economic pest in the south-eastern United States, has evolved practical resistance to Bacillus thuringiensis (Bt) Cry toxins in maize and cotton. Insect resistance management (IRM) programs have historically required planting of structured non-Bt maize, but because of its low adoption, the use of seed blends has been considered. To generate knowledge on target pest biology and ecology to help improve IRM strategies, nine field trials were conducted in 2019 and 2020 in Florida, Georgia, North Carolina, and South Carolina to evaluate the impact of Bt (Cry1Ab + Cry1F or Cry1Ab + Cry1F + Vip3A) and non-Bt maize plants in blended and structured refuge treatments on H. zea pupal survival, weight, soil pupation depth, adult flight parameters, and adult time to eclosion.

RESULTS

From a very large sample size and geography, we found a significant difference in pupal mortality and weight among treatments in seed blends with Vip3A, implying that cross-pollination occurred between Bt and non-Bt maize ears. There was no treatment effect for pupation depth, adult flight distance, and eclosion time.

CONCLUSION

Results of this study demonstrate the potential impact of different refuge strategies on phenological development and survival of an important pest species of regulatory concern. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

A DNA vaccine against dolphin morbillivirus is immunogenic in bottlenose dolphins

The immunization of exotic species presents considerable challenges. Nevertheless, for facilities like zoos, animal parks, government facilities and non-profit conservation groups, the protection of valuable and endangered species from infectious disease is a growing concern. The rationale for immunization in these species parallels that for human and companion animals; to decrease the incidence of disease. The U.S. Navy Marine Mammal Program, in collaboration with industry and academic partners, has developed and evaluated a DNA vaccine targeting a marine viral pathogen – dolphin morbillivirus (DMV). The DMV vaccine consists of the fusion (F) and hemagglutinin (H) genes of DMV. Vaccine constructs (pVR-DMV-F and pVR-DMV-H) were evaluated for expression in vitro and then for immunogenicity in mice. Injection protocols were designed for application in Atlantic bottlenose dolphins (Tursiops truncatus) to balance vaccine effectiveness with clinical utility. Six dolphins were inoculated, four animals received both pDMV-F and pDMV-H and two animals received a mock vaccine (vector alone). All animals received an inoculation week 0, followed by two booster injections weeks 8 and 14. Vaccine-specific immune responses were documented in all four vaccinated animals. To our knowledge, this is the first report of pathogen-specific immunogenicity to a DNA vaccine in an aquatic mammal species.     

Minimally‐invasive plate osteosynthesis of a Salter‐Harris type 2 fracture of the proximal phalanx in a filly

A 7‐month‐old Warmblood filly was referred to the equine hospital because of a Salter‐Harris type 2 fracture of the proximal physis of the right hind P1. On admission, the filly had acute grade 3/5 lameness at the walk and diffuse swelling in the area of the right hind proximal phalanx. Radiographs were taken to assess the fracture and plan treatment. Surgical treatment using two 4.5 mm narrow locking compression plates (PIP‐LCPs) resulted in fracture stabilisation and rapid healing. This case report suggests that internal fixation using PIP‐LCPs is useful for the treatment of Salter‐Harris type 2 fractures of the proximal physis of P1 in large foals.