Improved detection of Mycobacterium tuberculosis and M. bovis in African wildlife samples using cationic peptide decontamination and mycobacterial culture supplementation

In South Africa, mycobacterial culture is regarded as the gold standard for the detection of Mycobacterium tuberculosis complex (MTBC) infection in wildlife even though it is regarded as “imperfect.” We compared a novel decontamination and mycobacterial culture technique (TiKa) to the conventional mycobacterium growth indicator tube (MGIT) system using known amounts of bacilli and clinical samples from MTBC-infected African buffaloes (Syncerus caffer), white rhinoceros (Ceratotherium simum), and African elephants (Loxodonta africana). Use of the TiKa-KiC decontamination agent on samples spiked with 10,000 to 10 colony forming units (cfu) of M. bovis (SB0121) and M. tuberculosis (H37Rv) had no effect on isolate recovery in culture. In contrast, decontamination with MGIT MycoPrep resulted in no growth of M. bovis samples at concentrations < 1,000 cfu and M. tuberculosis samples < 100 cfu. Subsequently, we used the TiKa system with stored clinical samples (various lymphatic tissues) collected from wildlife and paucibacillary bronchoalveolar lavage fluid, trunk washes, and endotracheal tube washes from 3 species with known MTBC infections. Overall, MTBC recovery by culture was improved significantly (p < 0.01) by using TiKa compared to conventional MGIT, with 54 of 57 positive specimens versus 25 of 57 positive specimens, respectively. The TiKa mycobacterial growth system appears to significantly enhance the recovery of MTBC members from tissue and paucibacillary respiratory samples collected from African buffaloes, African elephants, and white rhinoceros. Moreover, the TiKa system may improve success of MTBC culture from various sample types previously deemed unculturable from other species.     

Comparison of hematologic and biochemical values for blood samples obtained via jugular venipuncture and via vascular access ports in cats

OBJECTIVE: To determine whether hematologic and serum biochemical values for blood samples obtained from cats via vascular access ports (VAP) are comparable to those for samples obtained by direct venipuncture. DESIGN: Prospective study. ANIMALS: 14 healthy cats. PROCEDURE: A VAP was surgically implanted in a jugular vein in each cat. Blood samples were obtained from the VAP and by direct venipuncture of the contralateral jugular vein 10 weeks after VAP placement. Results of hematologic and serum biochemical analyses were compared by use of a paired t-test. The Pvalue to reject the null hypothesis was adjusted to account for multiple comparisons by using the Bonferroni procedure in which the nominal P-to-reject value is divided by the number of comparisons (0.05/24 = 0.002). RESULTS: Paired samples (VAP and venipuncture) obtained 10 weeks after VAP placement were evaluated for each cat. Of the 24 measured analytes, only potassium, total protein, and albumin concentrations differed significantly (P< 0.001 for all 3) between VAP and venipuncture samples. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that samples obtained from VAP are suitable for routine hematologic monitoring of feline cancer patients. Sample hemolysis may account for a slight increase in potassium, total protein, and albumin concentrations obtained from VAP samples. However, the values of variables most critical for monitoring of patients receiving chemotherapy (ie, mature neutrophil and platelet counts) are comparable. If proper techniques are used, VAP may be used for administration of chemotherapy as well as for blood collection in cats undergoing cancer treatment.     

A Biophysical and Economic Assessment of a Community‐based Rehabilitated Gully in the Ethiopian Highlands

Gully erosion reduces agricultural productivity by destroying valuable land resources, increases sediment concentrations, reduces water quality, and fills up reservoirs. Gully rehabilitation has proven to be challenging especially in the high‐rainfall areas of the Ethiopian Highlands and has therefore had limited success. This paper describes a successful low‐cost gully rehabilitation effort with community participation in the Birr watershed in the Blue Nile basin that begun in early 2013. Initially, farmers were reluctant to participate for religious reasons, but with the aid of local priests and respected elders, community discussions, and a visit to a rehabilitated gully, a consensus was reached to rehabilitate a 0·71‐ha upland gully. The rehabilitation measures consisted of regrading the gully head at a 45° slope, constructing low‐cost check dams from locally available materials, and planting Pennisetum purpureum grass and Sesbania sesban. At the end of the first post‐implementation rainy season, 2,200 tons of soil was conserved by the constructed check dams and newly planted vegetation, compared with soil losses of 680 and 560 tons in two untreated, nearby gullies. In 2014, an additional 3,100 tons of soil was conserved. In 2013, the marginal rate of return (MRR) on the gully rehabilitation investment was 2·6 based on the value of increased forage production alone. When we include trapped soil nutrient values, the rehabilitation MRR was increased to 10. Although these numbers are impressive, the best proof of the success was that farmers on their own initiative rehabilitated an additional five gullies in 2014. Copyright © 2015 John Wiley & Sons, Ltd.     

Activation of the cellular proto-oncogene product p21Ras by addition of a myristylation signal

The 21-kD proteins encoded by ras oncogenes (p21Ras) are modified covalently by a palmitate attached to a cysteine residue near the carboxyl terminus. Changing cysteine at position 186 to serine in oncogenic forms produces a nonpalmitylated protein that fails to associate with membranes and does not transform NIH 3T3 cells. Nonpalmitylated p21Ras derivatives were constructed that contained myristic acid at their amino termini to determine if a different form of lipid modification could restore either membrane association or transforming activity. An activated p21Ras, altered in this way, exhibited both efficient membrane association and full transforming activity. Surprisingly, myristylated forms of normal cellular Ras were also transforming. This demonstrates that Ras must bind to membranes in order to transmit a signal for transformation, but that either myristate or palmitate can perform this role. However, the normal function of cellular Ras is diverted to transformation by myristate and therefore must be regulated ordinarily by some unique property of palmitate that myristate does not mimic. Myristylation thus represents a novel mechanism by which Ras can become transforming.     

Immunohistochemistry of adenosine deaminase: implications for adenosine neurotransmission

Immunohistochemical analysis of adenosine deaminase in rat brain revealed an extensive plexus of adenosine deaminase-containing neurons in the basal hypothalamus. These neurons converged on and were most numerous in three major centers, namely, the tuberal, caudal, and postmammillary caudal magnocellular nuclei. Most other brain regions were devoid of cells containing adenosine deaminase. Some adenosine deaminase-containing neurons were retrogradely labeled with the fluorescent dye fast blue when the dye was injected into the frontal cortex and striatum. Specific populations of neurons having high levels of adenosine deaminase may release adenosine as a neurotransmitter.     

Predicted and observed finger diameters in field soils

Wetting front instability resulting in fingered flow has been found in both wettable and nonwettable soils. Understanding how and when this phenomenon occurs under field conditions is greatly limited. Laboratory research has resulted in a number of expressions for finger diameter. In this paper we test the applicability of one of these equations for three different soils in the Netherlands where detailed soil sampling of moisture content was done earlier. In addition, information needed for finger prediction, such as the main wetting and drying loops of the soil moisture characteristic curves and the unsaturated and saturated soil conductivities, were measured in the laboratory. Results show that predicted finger diameters for the two sandy soils agreed well with the observed moisture patterns, while for the loess soil the wetting front was flat as predicted. The finger diameters in dry soil were based on the main wetting loop and in the wet soils were dependent on the main drying loop.     

Cardiopulmonary measurements in nonanesthetized, resting normal ponies.

Cardiopulmonary measurements were determined in 19 nonanesthetized, normal ponies. Mean values for arterial pressure, pulmonary arterial pressure, cardiac output, heart rate, packed cell volume, and hemoglobin are reported, as well as acid-base determinations of arterial blood and cerebrospinal fluid. Respiratory function test data include total ventilation, respiratory rate, alveolar ventilation, oxygen uptake, and carbon dioxide output. The data compare favorably with the available data from previous reports on ponies. Because of large day-to-day variations in total ventilation, alveolar ventilation should be measured if ponies are used in the study of pulmonary function.     

Genetic and Phenotypic Analysis of Soybean mosaic virus Resistance in PI 88788 Soybean

ABSTRACT Resistance to Soybean mosaic virus (SMV) was identified in PI 88788 soybean, a germ plasm accession from China that is used widely as a source of resistance to soybean cyst nematode. Strains SMV-G1 through -G7 infected the inoculated leaves of PI 88788 but were not detected in upper, noninoculated trifoliolate leaves. Inheritance of resistance was determined by inoculating progenies of crosses of PI 88788 with susceptible cvs. Essex and Lee 68 with SMV strains G1 and G7. Allelomorphic relationships with known genes for resistance to SMV were tested in crosses with the resistant genotypes PI 96983, L29, and V94-5152, possessing Rsv1, Rsv3, and Rsv4 genes, respectively. Data analyses showed that resistance in PI 88788 to SMV-G1 is controlled by a single, partially dominant gene; however, to SMV-G7, the same gene was completely dominant. The PI 88788 gene was independent of the Rsv1 and Rsv3 loci, but allelic to Rsv4 in V94-5152. Expression of the Rsv4 gene in PI 88788 resulted in a reduced number of infection sites and restricted short- and long-distance movement of virus, rather than hypersensitivity. A unique late susceptible phenotype was strongly associated with heterozygosity. This gene has potential value for use in gene pyramiding to achieve resistance to several SMV strains, as well as for rate-reducing resistance.     

Effect of staining and freezing media on sortability of stallion spermatozoa and their post-thaw viability after sex-sorting and cryopreservation

Sex‐sorted, frozen–thawed stallion spermatozoa remain out of reach of commercial horse breeders because of the low efficiency of the sex‐sorting process and unacceptable fertility rates after insemination. Two experiments were designed to test the effects of alternative staining and freezing media to improve the viability of sex‐sorted frozen–thawed stallion spermatozoa. Experiment 1 compared two freezing media, INRA 82^® and a modified lactose‐ethylenediaminetetraacetic acid (EDTA), for the cryopreservation of sex‐sorted stallion spermatozoa. No significant differences between the two freezing media could be identified, suggesting that both cryodiluents would be suitable for incorporation into a sex‐preselection protocol for stallion spermatozoa. Experiment 2 compared Kenney’s modified Tyrode’s (KMT) and Sperm TALP (Sp‐TALP) as the staining and incubation medium for stallion spermatozoa prior to sex‐sorting. A significant increase in the percentage of acrosome‐reacted spermatozoa occurred after staining and incubation in the clarified Sp‐TALP compared with KMT. As no improvements in sorting rates were achieved using Sp‐TALP, it was concluded that stallion sorting protocols could include KMT as the staining and incubation medium while either INRA 82^® or lactose‐EDTA could be employed as a cryodiluents.