Detection of hemorrhagic septicemia virus of salmonid fishes by use of an enzyme-linked immunosorbent assay containing high sodium chloride concentration and two noncompetitive monoclonal antibodies against early viral nucleoproteins.

Inclusion of high-ionic strength buffers helped us to develop a sandwich ELISA to detect hemorrhagic septicemia virus (HSV) in cell culture and infected trout tissue extracts. For maximal sensitivity of 0.1 to 0.2 ng/well/100 microliters or about 10 to 50 TCID50/well/100 microliters, trout extracts were diluted 1:1 and assayed for the earliest synthesized nucleoprotein N. Simultaneous binding of the N protein from HSV in the sample to the wells coated with monoclonal antibody (2D5 against the N protein) and to the peroxidase-labeled monoclonal antibody (2C9 against the N protein) proceeded during a 2-hour incubation at 20 to 22 C (room temperature). The response was linear between 6 to 60 ng/well of purified virus. Monoclonal antibodies used were noncompetitive with each other and reacted with F1, F2, 23.75, and 5 Spanish isolates of HSV, but not with infectious hematopoietic necrosis or infectious pancreatic necrosis viruses. Tissue specimens with low content of HSV virus may now be assayed directly without use of cell culture, rapidly, and with high precision, during the acute phase of the disease in salmonid fishes.     

Genotype and treatment biases in estimation of carcass lean of swine.

Carcasses of 181 barrows, representing five genotypes, 1) H x HD, 2) SYN, 3) HD x L[YD], 4) L x YD, and 5) Y x L (H = Hampshire, D = Duroc, SYN = synthetic terminal sire line, L = Landrace, and Y = Yorkshire), and two levels of ractopamine (RAC) treatment (0 and 20 ppm) were completely dissected and the data were used to examine genotype and treatment (RAC) biases in estimation of fat-standardized lean weight and to evaluate accuracies and precisions realized by use of equations based on variables derived from different technologies. Independent variables used to establish regression equations represented technologies of direct carcass measurements, optical probe data, TOBEC (total body electrical conductivity) readings, and dissected (DHMLN) and fat-standardized (FSHMLN) ham lean. Genotype bias existed when any equation from a single technology was used and was minimized by combining FSHMLN with one TOBEC reading, carcass length, and the probe measurement of 10th rib fat depth. Large RAC biases appeared when equations from direct carcass measurements or optical probe data were used and were minimized by an equation using either DHMLN or FSHMLN. A practical equation with relatively high R2 value and small genotype and RAC biases were developed by combining TOBEC readings with direct carcass measurements of 10th rib fat depth and warm carcass weight.     

A study on Theileria parva bovis carrier state

In two trials, Theileria parva bovis (which causes ‘January disease’ of cattle in Zimbabwe) produced a carrier state, over the 7–12 months after infection. Very severe clinical reactions were caused by infections from small numbers (29–43) of adult Rhipicephalus appendiculatus ticks, which had engorged on immunized cattle in the field. The transmission from healthy recovered cattle housed indoors was less efficient, even with high numbers of ticks (300). Two out of seven attempts were successful and disease reactions were rather severe. A non-pathogenic Theileria assumed to be Theileria taurotragi was transmitted in three out of seven attempts.     

Studies on the infundibular cysts of the uterine tube in camel (Camelus dromedarius).

Two hundred eighteen genital tracts of slaughtered female camels were collected and examined. Infundibular cysts were observed in 35 tracts (16%); these were either unilateral (22 cases) or bilateral (13 cases) all containing fluids of different consistencies. The morphological and histological structures of the cysts were recorded. The bacteriological investigation and physicochemical analysis of cyst contents were carried out. Aeromonas hydrophila was isolated from 68.5% of cases. Rectal palpation and ultrasound technique were compared for the diagnosis of the cysts antemortem.     

Detection of chromogranin A in the adrenal gland extracts of different animal species by an enzyme-linked immunosorbent assay using Thomsen-Friedenreich antigen-specific Amaranthus caudatus lectin

The reactivity of different lectins with crude chromogranin A (CgA) obtained from different animals, namely, cow, horse, dog, pig, and dolphin, was examined to identify lectin(s) that would be useful as coating reagent(s) in a sandwich enzyme-linked immunosorbent assay (ELISA). Of the different lectins studied, the Amaranthus caudatus lectin (ACA), which is specific for the Thomsen-Friedenreich (T)-antigen (Galβ1-3GalNAc), was found to react with the CgA from different animals by western blotting. Purified rabbit anti-bovine CgA antibody was also found to cross-react with the crude CgA preparations. On the basis of these findings, a sandwich ELISA was developed with ACA as the coating reagent and anti-bovine CgA antibody as the probing antibody. Using this method, concentration-dependent curves ranging from 0.003 μg/mL to 25 μg/mL and from 0.02 μg/mL to 25 μg/mL were obtained for bovine CgA and canine CgA, respectively. Similarly, concentration-dependent curves were obtained for the equine, swine, and dolphin crude CgA extracts. Thus, ACA is concluded to be a valuable reagent for CgA detection in crude extracts from different animal species, and for CgA isolation/purification.     

Adiposity and adiponectin in dogs: investigation of causes of discrepant results between two studies

Although one study showed lower adiponectin concentrations in obese dogs, other recent studies indicate that adiponectin might not be decreased in obese dogs, raising the possibility that the physiology of adiponectin is different in dogs than in humans. The aim of this study was to investigate possible causes of the discrepancy between the two largest studies to date that assessed the association between adiposity and adiponectin concentration in dogs, including the validity of the assay, laboratory error, and the effects of breed, sex, and neuter status on the relationship between adiposity and adiponectin concentrations. Adiponectin concentrations measured with a previously validated adiponectin ELISA were compared with those estimated by Western blotting analysis of reduced and denatured plasma samples. The possibility of laboratory error and the effect of EDTA anticoagulant and aprotinin were tested. Adiponectin concentration was measured by ELISA in 20 lean dogs (10 male and 10 female, 5 neutered in each sex). There was close correlation between adiponectin concentrations measured by ELISA and those estimated by Western blotting analysis (r = 0.90; P < 0.001). There was no substantial effect of EDTA, aprotinin, or laboratory error on the results. There was confounding by neuter status of the relationship between adiposity and adiponectin concentrations, but adiponectin concentrations were not significantly lower in male than in female lean dogs (females, 36 mg/L; males, 26 mg/L; P > 0.20) and were not significantly lower in intact than in neutered lean male dogs (intact, 28 mg/L; neutered, 23 mg/L; P = 0.49). We conclude that the adiponectin ELISA previously validated for use in dogs appears to be suitable for determination of canine adiponectin concentrations and that testosterone does not appear to have a strong effect on plasma adiponectin concentrations in dogs. Obesity might decrease adiponectin concentrations in intact but not in neutered dogs.     

Use of visible and near-infrared spectroscopy to predict pork longissimus lean color stability

This study evaluated the use of visible and near-infrared (VISNIR) spectroscopy to predict lean color stability in pork loin chops. Spectra were collected immediately after and approximately 1 h after rib removal on 1,208 loins. Loins were aged for 14 d before a 2.54-cm chop was placed in simulated retail display. Spectra were collected on aged loins immediately after removal from the vacuum package and on chops 10 min after cutting. Instrumental color measurements [L*, a*, b*, hue angle, chroma, and E (overall color change)] were determined on d 0, 1, 7, 11, and 14 of display. Principal components analysis of display d 0 and 14 values of these traits identified a factor (first principal component; PC1) explaining 67% of the variance that was related to color change. Partial least squares regression was used to develop 3 models to predict PC1 values by using VISNIR spectra collected in the plant, on aged loins, and on chops. Loins with predicted PC1 values less than 0 were classified as having a stable color, whereas values greater than 0 were classified as having a labile lean color. Loins classified as stable by the in-plant model had smaller (P < 0.05) L* values than those classified as labile. Hue angle and ΔE values were less (P < 0.05) and a* and chroma values were greater (P < 0.05) after d 7 of display in loins predicted to have a stable color than in loins predicted to have a labile lean color. Similarly, chops from loins classified as stable using the aged loin model had smaller (P < 0.05) L* values than those from loins classified as labile. Furthermore, loins predicted to be stable had smaller (P < 0.05) hue angle and ΔE values and greater (P < 0.05) a* and chroma values after d 7 of display than did loins predicted to be labile. Results for the chop model were similar to those from the 2 loin models. Chops predicted to have a stable lean color had smaller (P < 0.05) L* values than did those predicted to have a labile lean color. Chops classified as stable had smaller (P < 0.05) hue angle and ΔE values and greater (P < 0.05) a* and chroma values after d 7 of display compared with chops classified as labile. All 3 models effectively segregated chops based on color stability, particularly with regard to redness. Regardless of the model being used, d 14 display values for a*, hue angle, and ΔE in loins classified as stable were similar to the d 7 values of loins classified as labile. Thus, these results suggest that VISNIR spectroscopy would be an effective technology for sorting pork loins with regard to lean color stability.     

The major surface Vsp proteins of Brachyspira hyodysenteriae form antigenic protein complexes

The Vsp proteins are the major outer membrane proteins of Brachyspira hyodysenteriae, the causative agent of swine dysentery. Eight vsp genes have been identified in B. hyodysenteriae strain B204, arranged into two four-gene loci, and at least two of the corresponding proteins are produced in vitro. The aims of this study were to characterise the vsp genes of the virulent Australian B. hyodysenteriae strain X576 and their corresponding proteins, Genomic sequence comparison with strains B204 and WA1 demonstrated that the number of vsp genes varies between B. hyodysenteriae strains, although the chromosomal locations of the vsp gene loci are consistent. We identified two additional vsp-like genes, designated vspI and vspJ, in each of the three strains. Double SDS-PAGE was used to demonstrate that Vsp proteins of B. hyodysenteriae strain X576 form multimeric protein complexes in the outer membrane that are stable in 6M urea but dissociate after boiling. The Vsp complexes primarily consisted of VspF but also contain VspE and VspI. VspD was also found in a series of complexes slightly larger than the more abundant VspF complexes. Vsp proteins are purported to be antigenic; however little direct data are available to support this claim. In this study convalescent pig sera did not bind denatured Vsp proteins by Western blotting, but did bind the Vsp complexes on Western blots, showing that conformational epitopes may be important in immune recognition of these major outer membrane proteins. This is the first definitive demonstration of the antigenicity of these proteins in swine dysentery.