Abundant PrP(CWD) in tonsil from mule deer with preclinical chronic wasting disease.

A monoclonal antibody dot-blot assay was used to evaluate detergent lysates of tonsil tissue from mule deer to detect PrP(CWD), the marker for the cervid transmissible spongiform encephalopathy chronic wasting disease (CWD). Samples of formalin-fixed brain and tonsil tissues from mule deer were examined for PrP(CWD) using immunohistochemistry (IHC) with Mab F99/97.6.1, the gold standard for diagnosis of preclinical CWD. The contralateral tonsil from each of the 143 deer was prepared for confirmatory IHC and as a 10% (wt/vol) detergent lysate without purification or enrichment steps for monoclonal antibody dot-blot assay. PrP(CWD) was detected by dot-blot assay in 49 of 50 samples considered positive by IHC. Forty-eight of the positive samples were evaluated with a quantitative dot-blot assay calibrated with recombinant PrP. Tonsillar PrP(CWD) concentrations ranged from 34 to 1,188 ng per 0.5 mg starting wet weight of tissue. The abundant PrP(CWD) in mule deer tonsil will facilitate development and validation of high-throughput screening tests for CWD in large populations of free-ranging deer.     

Multiyear analysis of the dependency of the planting date on rainfall and soil moisture in paddy fields in Cambodia, 2003–2019

The dependencies of the planting date on rainfall and soil moisture in paddy fields in Cambodia were analyzed to quantify farmers’ empirical knowledge regarding their decision of the planting date. Remote sensing data from multiple satellites covering the 2003–2019 period were analyzed. The planting dates in rain-fed paddies ranged from April to August, with large spatial variations and year-to-year fluctuations. In years when planting was suppressed in April and May, planting was extensively enhanced in June and August compared to normal years, and vice versa. Over the northeastern side of Tonle Sap Lake and south of Phnom Penh city, the areas planted in April and May were found to have positive correlations with rainfall and soil moisture, suggesting that wetter-than-average conditions encouraged farmers to plant earlier in the season. In contrast, this relationship was unclear on the western side of Tonle Sap Lake, where the rainfall amounts were larger throughout the year than in other areas in Cambodia. In this region, the relationship between the planting area and soil water availability was either unclear or was even slightly negative from June to August. Since more frequent dry spells have been detected after the onset of the rainy season in recent years, further studies and disseminations of potential changes in dry spells are important for the agronomic adaptation of planting dates under climate change.

     

A comparison of Freund's Complete and Freund's Modified Adjuvants used with a contraceptive vaccine in wild horses (Equus caballus).

Fifteen captive wild mares (Equus caballus) were treated with porcine zona pellucida contraceptive vaccine and either Freund's Complete Adjuvant (n = 7) or Freund's Modified Adjuvant (n = 8). All mares received a booster inoculation of porcine zona pellucida plus Freund's Incomplete Adjuvant a month later. Anti-porcine zona pellucida antibodies were measured over 10 mo following the initial inoculation. There were no significant differences in antibody titers at any point during the 10 mo, and seven of the eight mares in the Freund's Modified Adjuvant group were above the 60% level at the end of the study, which is considered to be the contraceptive threshold for horses. There were no significant differences in titers between pregnant and nonpregnant horses, nor was there a significant correlation between age and titers. One local injection site reaction occurred after booster treatment with Freund's Incomplete Adjuvant, and 11 healthy foals were born during the course of the study. These data suggest that Freund's Modified Adjuvant is an acceptable substitute for Freund's Complete Adjuvant in certain free-ranging and captive wildlife species.     

Persistence of carbendazim in field-grown soybeans following application of thiophanate-methyl fungicide

Carbendazim was quantified spectrophotometrically in leaves, petioles, stems and pods from upper and lower halves of entire soybean plants following one or two foliar applications of thiophanate-methyl fungicide (1.1 kg ha^?1 a.i.). Following the first application, the highest carbendazim concentrations occurred 1-4 days post-application and declined thereafter to undetectable (?0.12 mg kg^?1) or barely detectable levels by day 26. The highest concentrations were in the leaves where carbendazim levels exceeded 3.5 mg kg^?1 on day 4 and remained above 1.0 mg kg^?1 until day 19. Concentrations in petioles and upper pods were initially in excess of 1.0 mg kg^?1 and remained above 0.5 mg kg^?1 for 12 days. In lower pods and in stems, concentrations were very low and often not detectable. Typically, concentrations were higher in samples from the upper half of the plant than in those from the lower half. A second application on day 18 resulted in carbendazim concentrations similar to those immediately after the first application; however, concentrations declined rapidly coincidentally with a period of rain. This decline may have been facilitated by poor fungicide uptake due to senescence of the plants.     

Mitochondrial DNA variation of the bocachico Prochilodus magdalenae (Characiformes,Prochilodontidae) in the Magdalena River Basin,Colombia

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Label-Free Darkfield-Based Technique to Assist in the Detection of <Emphasis Type="Italic">Giardia</Emphasis> Cysts

Giardia is a pathogenic protozoan that has caused giardiasis outbreaks worldwide, and this water-borne disease may occur due to faults in water supply and treatment systems. To support surveillance and control programs, the monitoring of this parasite is essential in water samples. Although Giardia cyst detection is usually based on standard light microscopy, the resulting low contrasted cysts together with a wide array of particles of comparable size to the target protozoan demands a high level of observer skill and experience, as well as a long time to process the individual samples. The visualization of this protozoan can be significantly improved by using staining procedures, e.g., Lugol’s iodine in brightfield or fluorescence-based methods such as fluorescence-isothiocyanate (FITC) and 4′,6-diamidino-2-phenylindole (DAPI). However, the significant costs make fluorescence microscopy difficult to be performed in less-developed countries. Accordingly, the present study evaluated the effectiveness of inexpensive darkfield (DF) setups to acquire higher contrasted images of unstained cyst samples as a direct and rapid method for monitoring protozoa. Two low-cost custom-made DF illumination modes, transmitted and reflected, were evaluated on a standard light microscope. Examination of purified Giardia cyst suspensions with both DF setups revealed a direct correlation between morphological appearance and uptake of DAPI. Images captured under transmitted illumination showed higher contrast and sharpness when compared to the reflected images. DF microscopy might provide a simple, direct, and inexpensive method for observing Giardia cysts, which shows basic aspects of their intracellular structure, although the applicability of the method to raw water concentrates remains to be demonstrated.