Development of a microtiter plate ELISA and a dipstick ELISA for the determination of the organophosphorus insecticide fenthion

In previous studies, polyclonal antibodies against the organophosphorus insecticide fenthion were obtained and an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed for this pesticide. In this study, using these antibodies and an enzyme tracer, direct competitive ELISAs for fenthion in microtiter plate and dipstick formats were developed. The microtiter plate ELISA showed an IC(50) value of 1.2 microg/L with a detection limit of 0.1 microg/L. The antibodies showed negligible cross-reactivity with other organophosphorus pesticides. The use of the dipstick format using Immunodyne as a support membrane allowed the quick visual detection of fenthion in concentrations >10 microg/L. The IC(50) value of the dipstick format using reflectance detection was 15 microg/L with a detection limit of 0.5 microg/L. The recoveries of fenthion from spiked vegetable samples using the two formats without any prior enrichment or cleanup steps were 87-116%.     

Single-nucleotide polymorphism-based epidemiological analysis of Korean Mycobacterium bovis isolates

BackgroundBovine tuberculosis (TB) is caused by Mycobacterium bovis, a well-known cause of zoonotic tuberculosis in cattle and deer, and has been investigated in many physiological and molecular studies. However, detailed genome-level studies of M. bovis have not been performed in Korea.ObjectivesTo survey whole genome-wide single-nucleotide polymorphism (SNP) variants in Korean M. bovis field isolates and to define M. bovis groups in Korea by comparing SNP typing with spoligotyping and variable number tandem repeat typing.MethodsA total of 46 M. bovis field isolates, isolated from laryngopharyngeal lymph nodes and lungs of Korean cattle, wild boar, and Korean water deer, were used to identify SNPs by performing whole-genome sequencing. SNP sites were confirmed via polymerase chain reaction using 87 primer pairs.ResultsWe identified 34 SNP sites with different frequencies across M. bovis isolates, and performed SNP typing and epidemiological analysis, which divided the 46 field isolates into 16 subtypes.ConclusionsThrough SNP analysis, detailed differences in samples with identical spoligotypes could be detected. SNP analysis is, therefore, a useful epidemiological tracing tool that could enable better management of bovine TB, thus preventing further outbreaks and reducing the impact of this disease.     

Systemic coccidioidomycosis in a llama cria native to Missouri

A 3-mo-old male llama was examined because of a 4-wk history of lethargy and ill thrift. Clinical examination revealed subcutaneous masses in the left prescapular and right inguinal regions, mild ataxia, a slight head tilt to the right, and right ear droop. The cria died before clinical workup was complete. At autopsy, there was generalized lymphadenomegaly, a hepatic nodule, a midbrain mass causing rostral compression of the cerebellum, and internal hydrocephalus. Microscopic findings included pyogranulomatous lymphadenitis, meningoencephalitis, hepatitis, and bronchopneumonia. Intralesional fungal spherules, most consistent with Coccidioides spp., were identified in the lymph nodes, lung, and brain. Fungal culture, single-nucleotide variation genotyping real-time PCR, and DNA sequencing confirmed Coccidioides posadasii. The dam of the cria was native to Arizona and had been moved to Missouri ~2.5 y previously. Agar gel immunodiffusion assay of the herd revealed that only the dam was positive for Coccidioides spp.; 6 herdmates were negative. Computed tomography of the dam revealed multiple nodules within the lungs and liver, which were presumed to be an active coccidioidomycosis infection. This case of systemic coccidioidomycosis in a llama native to Missouri was presumably acquired by vertical transmission from the dam.     

Dynamic in-lake spawning migrations by female sockeye salmon

Abstract –  Precise homing by salmon to natal habitats is considered the primary mechanism in the evolution of population-specific traits, yet few studies have focused on this final phase of their spawning migration. We radio tagged 157 female sockeye salmon ( Oncorhynchus nerka ) as they entered Lake Clark, Alaska, and tracked them every 1–10 days to their spawning locations. Contrary to past research, no specific shoreline migration pattern was observed (e.g., clockwise) nor did fish enter a tributary unless they spawned in that tributary. Tributary spawning fish migrated faster (mean = 4.7 km·day⁻¹, SD = 2.7, vs. 1.6 km·day⁻¹, SD = 2.1) and more directly (mean linearity = 0.8, SD = 0.2, vs. 0.4, SD = 0.2) than Lake Clark beach spawning fish. Although radio-tagged salmon migrated to within 5 km of their final spawning location in an average of 21.2 days (SD = 13.2), some fish migrated five times the distance necessary and over 50 days to reach their spawning destination. These results demonstrate the dynamic nature of this final phase of migration and support studies indicating a higher degree of homing precision by tributary spawning fish.     

Effect of yellow clay on respiration and phytoplankton uptake of bivalves

ABSTRACT:   Yellow clay dispersion has been applied to mitigate red tides in Korea since 1995. The present study documents the effect on physiology of two marine bivalves by examinations of clay uptake, respiration, and clearance rate ( CR ) of animals after the treatment of yellow clay. The amount of clay in gills and inside cavities of shellfish was increased with time, and the inside of the shellfish filled with clay in 2 h. The ejection of the accumulated clay was evident from as early as 30 min after removal from clay pools. Animals removed from the clay pool discharged all observable clay by 6 h. Within 30 min, CR decreased quickly and returned to the level of the untreated control groups within 2 h.     

Effects of breed, heterosis, age of dam, litter size, and birth weight on lamb mortality

Sources of individual plus maternal effects on lamb mortality were studied in data collected at the U.S. Meat Animal Research Center from 1980 to 1985 for paternal and maternal breeds lambing yearly and for maternal breeds lambing at 8-mo intervals. Records included 16,881 lambs born. Breeds included were Finnsheep (F), Dorset (D), Rambouillet (R), Suffolk (S), Targhee (T), Composite 1 (C1 = F/2, D/4, R/4), Composite 2 (C2 = F/2, S/4, T/4), and Composite 3 (C3 = Columbia/2, S/4, Hampshire/4), Traits analyzed were perinatal, postnatal, and total mortality to 60 d of age and postnatal respiratory, digestive, starvation, injury, and other or unknown causes of mortality. The least squares analyses included breed, year, sire within breed-year, sex, linear and quadratic season and age of dam covariates (Model 1), plus litter size (Model 2), plus birth weight (Model 3), and significant two-way interactions. Age of dam, litter size, and birth weight all had important, often quadratic, effects that differed among breeds. Models 1, 2, and 3, respectively, reduced variation by 8, 10, and 16% for perinatal, 7, 8, and 12% for postnatal, and 9, 11, and 20% for total mortality. In Model 1, breed means ranged from 3.5 to 16.2% for perinatal, 7.2 to 21.1% for postnatal, and 16.7 to 32.8% for total mortality. Respiratory and starvation problems were major causes of postnatal mortality. Heterosis for lamb survival retained in composites was 9% for C1 and 18% for C2. Mortality was 1 to 5% higher for males than for females.(ABSTRACT TRUNCATED AT 250 WORDS)     

Protection against Klebsiella pneumoniae respiratory tract infection of mice and squirrel monkeys given kanamycin by aerosol and injection.

The methods of aerosol administration of kanamycin and IM injection of the antibiotic were compared for their protection of mice and squirrel monkeys against Klebsiella pneumoniae respiratory tract infection. Mice exposed to LD90 of K pneumoniae at 0.5, 4, 24, 48 and 72 hours after they were treated with aerosol of kanamycin (27 mg/kg of body weight) were significantly better protected at all exposure times than were mice given the antibiotic (450 mg/kg) by IM injection. Squirrel monkeys given the aerosol at dose level of 11.25 mg/kg were completely protected against K pneumoniae exposure at 6 and 24 hours, whereas only one of eight monkeys treated with the same dose given IM survived the exposure at 6 hours and none survived at 24 hours. Antibiotic clearance curves indicated that kanamycin remained in the lungs at higher concentrations and for longer periods after aerosol treatment than after IM treatment.     

Mechanism of pressure-induced gelation of milk.

The pressure-induced gelation of concentrated skimmed milk and milk-sugar mixtures was studied to discover the main components responsible for gelation. The major protein component responsible for gelation is micellar casein. Gelation occurs at similar pressures to casein micelle disintegration in dilute milk, and both can be prevented by inclusion of excess calcium chloride. Transmission electron micrographs show that the protein network is formed from particles with diameters approximately an order of magnitude smaller than those of intact casein micelles. Gelation occurs on decompression and is found to be baroreversible. Concentrations of sugar up to 30% reduce the critical concentration of casein required for gelation, but higher sugar concentrations inhibit gelation. A mechanism of gelation based on the aggregation of casein submicelles formed by pressure-induced disintegration of casein micelles is proposed. Observations on the effect of sucrose on gelation are discussed in terms of the influence of sugars on the solvent quality in aqueous casein systems.