Precocene II metabolism: Comparative in vivo studies among several species of insects,and structure elucidation of two major metabolites

Several species of insects, exhibiting varying responsiveness to the juvenile hormone antagonist precocene II (6,7-dimethoxy-2,2-dimethylchromene), were challenged topically with a tritiated preparation of the title compound. Metabolism of [³H]precocene II was subsequently examined by withdrawing hemolymph samples from treated animals at appropriate time intervals and characterizing the extractable radiolabel chromatographically. Quantitative (or qualitative) differences observed between the respective metabolic profiles were found not correlative with specimen sensitivity to precocene. Production of two heretofore unreported metabolites, identified by spectral and chemical means as O-β-glucosides of 6- and 7-monodemethylated precocene II, was demonstrated in both sensitive and insensitive species. No evidence for the presence of a hemolymphborne, biologically effective “activated metabolite” produced in vivo by precocene-susceptible insects could be found. The latter finding may well argue for in situ bioactivation of precocene at the target tissue(s) by these sensitive insects.     

Evaluation of iatrogenic hemarthrosis of the metacarpophalangeal joint as a method of induction of temporary reversible lameness in horses

OBJECTIVE: To determine whether iatrogenic hemarthrosis of the metacarpophalangeal joint could be used as a model for temporary reversible joint pain in horses. ANIMALS: 8 adult horses. PROCEDURE: Each horse was evaluated on a treadmill before and after injection of 1 metacarpophalangeal joint with 10 mL of autogenous blood. Horses were evaluated subjectively and objectively by use of a computerized force measurement system at intervals until lameness abated. The mean force difference between injected and noninjected limbs at all time periods after injection was compared with the difference between limbs at baseline. From each horse, synovial fluid samples collected before and 24 hours and 30 days after injection were analyzed for total protein concentration and cell type and number. Venous blood samples were collected before and 6 and 24 hours after injection for assessment of plasma cortisol concentration. RESULTS: For 24 hours after injection, the mean force difference between injected and noninjected limbs was significantly increased over baseline. The greatest force difference was detected after 2 and 4 hours. Baseline and 24-hour force data were not significantly different. Compared with baseline values, synovial fluid protein concentration and nucleated cell and RBC counts were increased significantly at 24 hours after injection but were not different at 30 days after injection. No significant changes in plasma cortisol concentration were detected at any time point. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, iatrogenic hemarthrosis of the metacarpophalangeal joint appears to induce temporary reversible lameness with a mild to moderate degree of synovitis.     

Evaluation of three enzyme-linked immunosorbent assays (ELISAs) for the detection of serum antibodies in sheep infected with Echinococcus granulosus

The aim of this study was to develop an immunological method for the identification of sheep infected with Echinococcus granulosus which would allow the monitoring of animals imported into countries free from hydatidosis and as an aid to countries where control schemes for the disease are in operation. Three enzyme-linked immunosorbent assays (ELISAs) were developed and validated, using as antigen either a purified 8 kDa hydatid cyst fluid protein (8kDaELISA), a recombinant EG95 oncosphere protein (OncELISA) or a crude protoscolex preparation (ProtELISA). Sera used for the assay validations were obtained from 249 sheep infected either naturally or experimentally with E. granulosus and from 1012 non-infected sheep. The highest diagnostic sensitivity was obtained using the ProtELISA at 62.7 and 51.4%, depending on the cut-off. Assay sensitivities were lower for the 8kDaELISA and the OncELISA. Diagnostic specificities were high, ranging from 95.8 to 99.5%, depending on the ELISA type and cut-off level chosen. A few sera from 39 sheep infected with T. hydatigena and from 19 sheep infected with T. ovis were recorded as positive. Western immunoblot analysis revealed that the dominant antigenic components in the crude protoscolex antigen preparation were macromolecules of about 70-150 kDa, most likely representing polysaccharides. This study demonstrated that the ProtELISA was the most effective immunological method of those assessed for detection of infection with E. granulosus in sheep. Because of its limited diagnostic sensitivity of about 50-60%, it should be useful for the detection of the presence of infected sheep on a flock basis and cannot be used for reliable identification of individual animals infected with E. granulosus.     

Stratum corneum removal facilitates experimental sensitization to mite allergens in atopic dogs

In humans with atopic dermatitis and in mouse models of IgE-mediated allergic diseases, evidence is mounting that the stratum corneum (SC) provides an important barrier against environmental allergens. At this time, it is not known whether the SC has a similar role in dogs, especially in those with atopic dermatitis. The objectives of this pilot study were to determine whether SC removal led to earlier and stronger sensitization of atopic dogs to Dermatophagoides farinae (Df) house dust mites. Five Maltese-beagle atopic (MBA) dogs were sensitized epicutaneously after the SC was removed with ten tape strips (TS group), while sensitization was done without tape strips in five other MBA dogs (nontape stripping; NTS group). During this 16 week study, sensitization was assessed with allergen-specific IgE serology, intradermal testing with Df allergens and determination of stimulation indices of blood mononuclear cells cultured with Df and stained for CD4 and the activation markers CD25 or CD30. Compared with dogs from the NTS group, those of the TS group exhibited earlier rises in Df-specific IgE serum levels, usually had higher allergen-specific IgE titres, showed higher intradermal test reactivity and had earlier increases and higher percentages of CD25- or CD30-positive activated allergen-specific peripheral CD4-positive T lymphocytes. These observations implicate a role of the SC as a barrier limiting sensitization to exogenous allergens in this experimental atopic dog model.     

Osteomyelitis of the pelvis caused by Rhodococcus equi in a two-year-old horse

A 2-year-old female Quarter Horse was evaluated for hind limb lameness, fever (40 degrees C [104 degrees F]), and lethargy of 2 weeks' duration. Hypoproteinemia characterized by hypoalbuminemia and hyperfibrinogenemia was detected. Abdominal ultrasonography revealed thickening of the right dorsal colon wall. Treatment was instituted for putative right dorsal coliis. Lameness evaluation localized signs of pain to the lumbar vertebrae or pelvis. Radiography performed with the horse standing and nuclear scintigraphy revealed no abnormalities. Ventrodorsal pelvic radiography revealed a focal area of bony lysis and proliferation involving the cranial portion of the pubic symphysis. Aspiration yielded purulent exudate containing Rhodococcus equi, which was susceptible to erythromycin. Treatment included surgical debridement of the abscess and oral administration of erythromycin and rifampin. The horse's hind limb lameness completely resolved within 20 days. Infections of the pubic symphysis should be considered when lameness localized to the pelvis is associated with fever and an inflammatory leukogram.     

DISTAL BORDER FRAGMENTS OF THE EQUINE NAVICULAR BONE: ASSOCIATION BETWEEN MAGNETIC RESONANCE IMAGING CHARACTERISTICS AND CLINICAL LAMENESS

Distal border fragments of the navicular bone are increasingly being detected due to the improved capabilities of magnetic resonance imaging (MRI), but their clinical significance remains unclear. The purpose of this retrospective study was to describe the location, size, and frequency of fragments in a cohort of horses presented for MRI of the foot and to compare MRI findings with severity of lameness. Archived MRI studies and medical records were searched from March 2006 to June 2008. Horses were included if a distal border fragment of the navicular bone was visible in MRI scans. Confidence interval comparisons and linear regression analyses were used to test hypotheses that fragments were associated with lameness and lameness severity was positively correlated with fragment volume and biaxial location. A total of 453 horses (874 limbs) were included. Fragments were identified in 60 horses (13.25%) and 90 limbs (10.3%). Fifty percent of the horses had unilateral fragments and 50% had bilateral fragments. Fragments were located at the lateral (62.2%), medial (8.89%), or medial and lateral (28.9%) angles of the distal border of the navicular bone. There was no increased probability of being categorized as lame if a fragment was present. There was no significant difference in fragment volume across lameness severity categorizations. Confidence intervals indicated a slightly increased probability of being classified as lame if both medial and lateral fragments were present. Findings indicated that distal border fragments of the navicular bone in equine MRI studies are unlikely to be related to existing lameness.     

A systematic review and meta‐analysis of predictors of human hepatitis E virus exposure in non‐endemic countries

The reported incidence of clinical hepatitis E cases is rising in some non‐endemic countries, with concurrent concerns regarding potential hepatitis E virus (HEV) contamination of the blood supply. Therefore, the characterization of major potential sources of human HEV exposure is important to inform risk assessment and public health policy. A systematic review was conducted, including a comprehensive search in six electronic bibliographic databases, verified by hand‐searching reference lists of HEV reviews, and a grey literature search, of the broad research question ‘what is the evidence of the association between predictors of human HEV exposure, and HEV IgG seropositivity, in non‐endemic countries?’ Using forms designed a priori, captured studies were appraised at first‐level screening, second‐level characterization, and third‐level data extraction and risk of bias assessment. Meta‐analysis yielded summary estimates of association between potential predictors and odds of HEV seropositivity. Meta‐analysis and meta‐regression of the odds of HEV seroprevalence in specific groups characterized potential sources of HEV exposure. From 4,163 captured citations, 245 relevant studies underwent data extraction, investigating HEV seroprevalence or predictors in both healthy subjects and targeted patient groups. Across these groups, increasing age was a predictor of HEV IgG seropositivity. Both human immunodeficiency virus patients and haemodialysis patients had significantly increased odds of HEV seropositivity relative to the general population. Working with pigs, in forestry, or in hospitals, was significantly associated with increased odds of HEV seropositivity, as were consumption of meat, pork or game meat, or hunting. Chronological time was not associated with HEV seropositivity within our data sets. Further study of the distribution of potential dietary or behavioural predictors between high and lower prevalence areas within non‐endemic countries could improve our understanding of the relative importance of specific HEV transmission pathways.     

Comparative immunogenicity of M. hyopneumoniae NrdF encoded in different expression systems delivered orally via attenuated S. typhimurium aroA in mice

The Mycoplasma hyopneumoniae ribonucleotide reductase R2 subunit (NrdF) gene fragment was cloned into eukaryotic and prokaryotic expression vectors and its immunogenicity evaluated in mice immunized orally with attenuated Salmonella typhimurium aroA CS332 harboring either of the recombinant expression plasmids. We found that NrdF is highly conserved among M. hyopneumoniae strains. The immunogenicity of NrdF was examined by analyzing antibody responses in sera and lung washes, and the cell-mediated immune (CMI) response was assessed by determining the INF-gamma level produced by splenocytes upon in vitro stimulation with NrdF antigen. S. typhimurium expressing NrdF encoded by the prokaryotic expression plasmid (pTrcNrdF) failed to elicit an NrdF-specific serum or secretory antibody response, and IFN-gamma was not produced. Similarly, S. typhimurium carrying the eukaryotic recombinant plasmid encoding NrdF (pcNrdF) did not induce a serum or secretory antibody response, but did elicit significant NrdF-specific IFN-gamma production, indicating induction of a CMI response. However, analysis of immune responses against the live vector S. typhimurium aroA CS332 showed a serum IgG response but no mucosal IgA response in spite of its efficient invasiveness in vitro. In the present study we show that the DNA vaccine encoding the M. hyopneumoniae antigen delivered orally via a live attenuated S. typhimurium aroA can induce a cell-mediated immune response. We also indicate that different live bacterial vaccine carriers may have an influence on the type of the immune response induced.     

Milk antibodies against Ostertagia ostertagi: relationships with milk IgG and production parameters in lactating dairy cattle

The present study was carried out to evaluate the relationship between milk optical density ratios (ODRs) from an indirect Ostertagia ostertagi ELISA, total milk IgG levels and milk production and then establish a correction factor to adjust ODR. Five hundred and sixty composite milk samples collected from 358 cows on four dairy herds in June and August 2002 were used in this analysis. The average ODR was 0.34. A positive correlation was found between ODR and IgG values in milk, days in milk, age and log transformed somatic cell counts (SCC). However, ODR was negatively correlated with milk production. The IgG levels and ODR values were constant from 30 to 200 days in milk. However, ODRs increased from 200 days until the end of the lactation. After controlling for age, season, herd and SCC, an increase in milk production of 13 kg/day was associated with a reduction in ODR values of 0.052. The results of the present study suggest that ODR values are not greatly influenced by production factors. ODR follow the same pattern as the IgG variation across lactation and could be adjusted in order to compare ODR values obtained from high producing cows with those obtained from low producing animals.