Prevalence of Mycobacterium paratuberculosis in ileocecal lymph nodes of cattle culled in the United States

During 1983 and 1984, ileocecal lymph node specimens were obtained from clinically normal cattle at 76 US Department of Agriculture-inspected cull cattle slaughterhouses in 32 states and Puerto Rico. Mycobacterium paratuberculosis was isolated from the lymph nodes of 119 of 7,540 cattle. The prevalence of bovine paratuberculosis was 1.6% overall, with 2.9% in dairy culls and 0.8% in beef culls. The prevalence for females and males did not appear to differ significantly.     

Resistance in Lycopersicon esculentum Intraspecific Crosses to Race T1 Strains of Xanthomonas campestris pv. vesicatoria Causing Bacterial Spot of Tomato

ABSTRACT We used molecular markers to identify quantitative trait loci (QTL) that confer resistance in the field to Xanthomonas campestris pv. vesicatoria race T1, a causal agent of bacterial spot of tomato. An F(2) population derived from a cross between Hawaii 7998 (H 7998) and an elite breeding line, Ohio 88119, was used for the initial identification of an association between molecular markers and resistance as measured by bacterial populations in individual plants in the greenhouse. Polymorphism in this cross between a Lycopersicon esculentum donor of resistance and an elite L. esculentum parent was limited. The targeted use of a core set of 148 polymerase chain reaction-based markers that were identified as polymorphic in L. esculentum x L. esculentum crosses resulted in the identification of 37 markers that were polymorphic for the cross of interest. Previous studies using an H 7998 x L. pennellii wide cross implicated three loci, Rx1, Rx2, and Rx3, in the hypersensitive response to T1 strains. Markers that we identified were linked to the Rx1 and Rx3 loci, but no markers were identified in the region of chromosome 1 where Rx2 is located. Single marker-trait analysis suggested that chromosome 5, near the Rx3 locus, contributed to reduced bacterial populations in lines carrying the locus from H 7998. The locus on chromosome 5 explained 25% of the phenotypic variation in bacterial populations developing in infected plants. An advanced backcross population and subsequent inbred backcross lines developed using Ohio 88119 as a recurrent parent were used to confirm QTL associations detected in the F(2) population. Markers on chromosome 5 explained 41% of the phenotypic variation for resistance in replicated field trials. In contrast, the Rx1 locus on chromosome 1 did not play a role in resistance to X. campestris pv. vesicatoria race T1 strains as measured by bacterial populations in the greenhouse or symptoms in the field. A locus from H 7998 on chromosome 4 was associated with susceptibility to disease and explained 11% of the total phenotypic variation. Additional variation in resistance was explained by plant maturity (6%), with early maturing families expressing lower levels of resistance, and plant habit (6%), with indeterminate plants displaying more resistance. The markers linked to Rx3 will be useful in selection for resistance in elite x elite crosses.     

Serum complement activity and serum enzymes in rats after a subcutaneous injection of toxin prepared from Pasteurella multocida type D

Toxin produced by Pasteurella multocida type D was investigated for its effect on serum complement and serum biochemistry in rats. Rats were given a sublethal single subcutaneous injection of D toxin equivalent to 0.2 microgram/kg of body weight. Serum obtained 1, 3, 5 and 7 days post-treatment was tested for complement activity, total bilirubin, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP). Serum complement titers were significantly elevated (P less than 0.05) at all times after injection of toxin compared to rats injected with diluent and tested at the same intervals. Bilirubin was decreased but both control and D toxin-treated rats had low concentrations of bilirubin in their sera. The other biochemical constituents measured had no consistent pattern that would indicate liver damage in the rats.     

Evaluation of field isolates of pseudorabies (Aujeszky's disease) virus as determined by restriction endonuclease analysis and hybridization

Sixty-seven isolates of pseudorabies virus (PRV) from 13 states were cleaved with 4 restriction endonucleases (RE), and after electrophoresis in agarose, their banding patterns were photographed and evaluated. The deoxyribonucleic acid (DNA) cleavage fragments were designated into regions specified by molecular weight ranges based on lambda phage DNA as a size marker. The 67 PRV isolates were evaluated according to the total number of cleavage fragments, by the number of fragments within designated molecular weight regions, and finally, by the migration of fragments within regions. Four of the 67 PRV isolates (all 4 from California) did not have a 4.1 to 4.6 megadalton HinfI band, but hybridization of the HinfI digests with a 32P probe made with the 4.4 megadalton band hybridized with 2 lighter fragments, 2.5 and 1.9 megadaltons, respectively. The BamHI digests of DNA from some PRV isolates with submolar fragments were hybridized with 32P probes made with fragments from the submolar region and the BamHI E fragment. Both probes hybridized to the submolar region of PRV with BamHI submolar fragments, but only to the trimolar (E, F, and G) band of PRV without submolar fragments in the 4.1 to 7.5 BamHI megadalton region. Epidemiologic evidence was obtained which indicated that a Missouri strain of PRV was transferred to an Illinois swine herd by importation of feeder pigs from Missouri. The results indicate that there are numerous genomically different PRV currently in circulation in the United States and that the combination of RE analysis and DNA hybridization offers useful epidemiologic information to evaluate the various strains.     

Effect of muramyl dipeptide on immunogenicity of Corynebacterium pseudotuberculosis whole-cell vaccines in mice and lambs

Colostrum-deprived lambs and CF1 mice were vaccinated with water-in-oil emulsion vaccines containing nonviable whole cells (WC) of Corynebacterium pseudotuberculosis with and without muramyl dipeptide (MDP). Efficacy of vaccines was determined from the survival of mice and lesions in lambs after IV injection of 10(4) colony-forming units of C pseudotuberculosis. In mice, protection was related to the concentration of WC in the vaccine. At 50, 100, or 150 micrograms of WC, protection was good (78.8%). At 10 or 25 micrograms of WC, protection was considerably less (54.7%). At high WC concentrations, protection could only be moderately increased to 82.3% with high (50 and 100 micrograms) concentrations of MDP or increased to 90% protection with low (5 and 10 micrograms) concentrations of MDP. At low WC concentrations, protection significantly decreased to 32% (P less than 0.025) with high concentrations of MDP, but significantly increased to 72.5% (P less than 0.025) with low concentrations of MDP. Therefore, the amount of protection with lower concentrations of WC and MDP was comparable with the amount of protection with higher concentrations of WC without MDP. In lambs, high prechallenge antibody titers (geometric mean titers from 5.1 to 5.4 by day 35) were observed after vaccination with WC. Protection and vaccination site abscesses in lambs were related to the concentration of WC and MDP. Pulmonary or vaccination site abscesses were not observed in 4 of 4 lambs vaccinated with 1 mg of WC + 50 micrograms of MDP.     

Severe Leukemoid Response Associated With Mycobacterium genavense Infection in a Pet Budgerigar (Melopsittacus undulatus)

A 2-year-old pet budgerigar was presented after a period of general malaise that was unresponsive to supportive therapy. Radiographic images revealed a marked hepatomegaly. Hematology results indicated the bird had a severe leukocytosis with marked discrepancy between the hemocytometer white blood cell (WBC) count and WBC estimated from the smear. The leukocytosis was characterized by an extreme heterophilia with marked left shift, including the presence of metamyelocytes and promyelocytes with marked toxic changes, and a marked monocytosis, consistent with a leukemoid response. The bird died despite supportive treatment. Hepatosplenomegaly was present at necropsy. Histopathology of multiple tissues demonstrated acid-fast organisms on Ziehl-Neelsen staining, subsequently identified as Mycobacterium genavese by polymerase chain reaction analysis. Mycobacteriosis should be considered as a differential diagnosis in the presence of a severe leukemoid response with multiple heterophilic precursors in avian species.     

Coffee and liver diseases

Coffee consumption is worldwide spread with few side effects. Interestingly, coffee intake has been inversely related to the serum enzyme activities gamma-glutamyltransferase, and alanine aminotransferase in studies performed in various countries. In addition, epidemiological results, taken together, indicate that coffee consumption is inversely related with hepatic cirrhosis; however, they cannot demonstrate a causative role of coffee with prevention of liver injury. Animal models and cell culture studies indicate that kahweol, diterpenes and cafestol (some coffee compounds) can function as blocking agents by modulating multiple enzymes involved in carcinogenic detoxification; these molecules also alter the xenotoxic metabolism by inducing the enzymes glutathione-S-transferase and inhibiting N-acetyltransferase. Drinking coffee has been associated with reduced risk of hepatic injury and cirrhosis, a major pathogenic step in the process of hepatocarcinogenesis, thus, the benefit that produces coffee consumption on hepatic cancer may be attributed to its inverse relation with cirrhosis, although allowance for clinical history of cirrhosis did not completely account for the inverse association. Therefore, it seems to be a continuum of the beneficial effect of coffee consumption on liver enzymes, cirrhosis and hepatocellular carcinoma. At present, it seems reasonable to propose experiments with animal models of liver damage and to test the effect of coffee, and/or isolated compounds of this beverage, not only to evaluate the possible causative role of coffee but also its action mechanism. Clinical prospective double blind studies are also needed.     

Introgression of resistance to reniform nematode (Rotylenchulus reniformis) into upland cotton (Gossypium hirsutum) from Gossypium arboreum and a G. hirsutum/Gossypium aridum bridging line

Gossypium hirsutum L. is the predominant cotton of commerce and all cultivars of this species are susceptible to the reniform nematode, Rotylenchulus reniformis. To introgress resistance to R. reniformis into the tetraploid 2(AD₁) G. hirsutum, a resistant diploid A₂-genome Gossypium arboreum accession (A₂-190) was crossed with a hexaploid 2((AD₁)D₄) bridging line (G 371) to obtain a tetraploid triple-species hybrid. The triple-species hybrid was back-crossed to G. hirsutum and a population of 277 BC₁ individuals was produced. The BC₁s and controls were assayed in growth chambers for resistance to R. reniformis. Fortuitously, the hexaploid bridging line G 371 was also found to be resistant to R. reniformis. The BC₁ segregated 3:1, resistant:susceptible, indicating that resistance was conferred by dominant genes at two different loci, with each originating from a distinct germplasm source. This study demonstrated that it is possible to introgress and pyramid genes for resistance to R. reniformis in G. hirsutum.     

Morphometric evaluation of changes in the digestive tract of rainbow trout (Oncorhynchus mykiss) due to fish meal replacement with soy protein concentrate

Effects of the complete replacement of fish meal (FM) by soy protein concentrate (SPC) in high-energy diets (23 MJ kg^− 1 gross energy) on intestinal and liver histology were investigated in rainbow trout (Oncorhynchus mykiss). We developed a novel morphometric method to quantify the changes at the macro- and micromorphology level. Fish were acclimatized to the diet for 90 d and were then fed 1.0 g kg BW^− 1 d^− 1 once per day for 14 d before 10 fish per diet were randomly sampled, 6 h after their last meal. Fold height, epithelium length, stroma proportion of the proximal and distal intestines were not affected by the diet. No significant diet effect was detected on the features of the valve in the distal intestine. Fold height was linked to epithelium length in the proximal and distal intestines and this relationship was not modified by FM replacement. Enterocyte height and width were lower in SPC-fed fish in the distal intestine, but in the proximal intestine their morphology was not affected by the diet. There was no diet effect on the number of cells infiltered between the enterocytes in the proximal and distal intestines. The enterocyte nucleus was closer to the apex in the proximal and distal intestines in SPC-fed fish, but the features of the nucleus were not affected by the diet. The mean hepatocyte volume was on an average 36% lower in SPC-fed fish and was positively correlated to the hepatosomatic index for fish fed this diet only. In conclusion, SPC used in the present study did not either cause inflammatory reaction of the gut nor affected the epithelium surface. We suggest that the modifications due to SPC are linked to variations in the metabolic status of the enterocyte.     

Demonstration and validation of enhanced monitored natural recovery at a pesticide-contaminated sediment site

Purpose

Monitored natural recovery (MNR) combined with a thin-layer cap (TLC), often referred to as enhanced monitored natural recovery (EMNR), has the potential to accelerate and improve the effectiveness of MNR as a remedial strategy while minimizing widespread disturbance to the existing habitat. This study evaluated the effectiveness of a nominal 15-cm thin-layer sand cap as an EMNR remedial strategy to address sediments that were moderately contaminated with the chlorinated pesticide dichlorodiphenyltrichloroethane and its derivatives, collectively DDX.

Materials and methods

Physical, chemical, and biological measurements were conducted pre-remedy placement and 2, 14, and 25 months post-placement. Measurements were used to evaluate (1) TLC stability; (2) bottom-up mixing of the TLC; (3) advection through the TLC; (4) characteristics of newly deposited sediment atop the EMNR layer compared to pre-remedy surface sediment conditions; (5) changes in contaminant bioavailability; and (6) physical impacts to the benthic community.

Results and discussion

Significant reductions were observed from measurements conducted pre- and post-placement in surface sediment (84–97%), porewater (33–75%), and tissue concentrations (Lumbriculus variegatus deployed in situ) (72–82%). A 63 to 72% decrease in DDX depositional mass flux also was observed. Multiple lines of evidence indicated that the TLC material remained stable. Deposition of suspended material with low concentrations of DDX influenced low concentrations in the surface sediments. No adverse effects were observed on the benthic invertebrate community after TLC placement, and ecological metrics indicated increases in benthic community health, even in the short time period (2 months) following TLC placement.

Conclusions

This demonstration showed that EMNR can be effective at reducing biological exposure in surface sediments while minimizing short-term disturbances to benthic communities at sites where MNR is a remedy option, but natural deposition rates are inadequate to achieve cleanup goals within a reasonable timeframe.