Second generation competitive enzyme immunoassay for detection of bovine antibody to Brucella abortus

A second generation competitive enzyme immunoassay (CELISA) for detection of bovine antibody to Brucella abortus was developed. This assay was different from previously developed CELISAs in that the detection reagent used was a recombinant combination of the receptor portions of protein A and protein G, labelled with horseradish peroxidase. This eliminates the need for polyclonal anti-mouse-enzyme conjugate reagents for detection thus allowing for true standardization. The assay utilized a monoclonal antibody specific for a common epitope of the O-polysaccharide (OPS) of smooth lipopolysaccharide (SLPS) derived from B. abortus S1119.3 but which did not react with protein A/G. This monoclonal antibody was used to compete with antibody in the bovine test serum. Binding of bovine antibody to the smooth lipopolysaccharide antigen was then measured directly with the protein A/G enzyme conjugate. In this case, development of colour in the reaction was indicative of the presence of bovine antibody. The performance characteristics, sensitivity, specificity and exclusion of B. abortus S19 vaccinated animals, of the assay were very similar to those of the classical CELISA.     

Follicular,Oocyte and Embryo Features Related to Metabolic Status in Primiparous Lactating does Fed with High‐Fibre Rearing Diets

Fertility of primiparous lactating does in the early postpartum (pp) period is very low mainly due to pronounced deficient energy intake, influencing oocyte and embryo developmental competence. The hypothesis used in this work was that high‐lignin fibre diet supplied during the rearing period could increase feed intake and, consequently, improve the reproductive physiology and metabolic status of primiparous does in the early pp period. Diets with high‐lignin [HL: 15.8% dry matter (DM)] or standard‐lignin content (SL: 4.9% DM) were supplied until parturition time. No diet effects in serum oestradiol, progesterone concentrations and follicle categories were found in the histological study. Metaphase II rate of in vitro‐matured oocytes was significantly higher in the SL vs the HL group (p < 0.001). Cytoplasmically degenerated oocytes (in terms of abnormal distribution of cortical granules) and follicular atresia rate were significantly lower in the SL group than in the HL group (p < 0.05 and p < 0.005 respectively). In addition, HL‐fed does showed lower number of viable embryos and higher rate of retarded in vivo‐recovered embryos compared with the SL group (p < 0.05). Neither in vitro embryo development of viable embryos nor conception rate was significantly different between groups. Feed intake increased during the first pregnancy in the HL group (p < 0.05), but not during early lactation. Serum protein, non‐esterified fatty acid and leptin concentrations, as well as estimated body composition were similar in does fed with both diets. In conclusion, the enhancement of reproductive management by using highly lignified products in rearing diets does not seem to report physiological reproductive benefits affecting oocyte maturation rate and embryo viability in primiparous lactating does.     

Evaluating top soil trace element pollution in the vicinity of a cement plant and a former open-cast uranium mine in central Argentina

Purpose  

Heavy metals are especially dangerous because of their persistence and toxicity. Soil behaves as a sink of heavy metals by aerial deposition of particles emitted by different human activities. The aims of this work were to identify the levels and sources of heavy metal and trace elements in agricultural and residential areas in Argentina and to evaluate the enrichment of total and HCl-extracted heavy metals.     

Validation of a second generation competitive enzyme immunoassay (CELISA) for the diagnosis of brucellosis in various species of domestic animals

A second generation competitive enzyme immunoassay (CELISA) for detection of bovine antibody to Brucella abortus was developed to eliminate reagent variables in the assay. This assay was different from earlier CELISA formats in that it used recombinant protein A and protein G immunoglobulin receptors (PAG), labelled with horseradish peroxidase, thus eliminating the requirement for polyclonal anti-mouse-enzyme conjugate for detection. This allowed standardization of the assay. The CELISA uses a monoclonal antibody specific for a common epitope of the O-polysaccharide (OPS) of smooth lipopolysaccharide (SLPS) derived from B. abortus S1119.3. This antibody did not react with PAG. This monoclonal antibody was used to compete with antibody in the bovine test serum to the smooth lipopolysaccharide (SLPS) antigen. Reaction of bovine antibody was then measured directly with the PAG enzyme conjugate. In this case, development of colour in the reaction indicated a positive reaction. The performance characteristics of the new CELISA, sensitivity, specificity and exclusion of antibody of B. abortus S19 vaccinated animals, were very similar to those of the classical CELISA and to the indirect enzyme immunoassay (IELISA) when using sera deemed positive by isolation of the bacterium, either from individual animals or from some animals on the premises. All sera were tested by the buffered antigen plate agglutination test (BPAT) and the complement fixation test (CFT). Only samples positive on both BPAT and CFT were considered as positive and only samples negative on both tests were used considered negative. Sufficient samples from cattle, swine, sheep and goats to validate the test were included based on OIE guidelines suggesting inclusion of a minimum of 300 positive and 1000 negative samples.     

Prospective evaluation of pancarpal arthrodesis for carpal injuries in working dogs in New Zealand,using dorsal hybrid plating

Abstract

AIM: To determine whether working dogs in New Zealand with carpal injuries and treated with unilateral pancarpal arthrodesis (PCA), using a dorsal hybrid-plating method, are able to return to satisfactory working ability.

METHODS: Fourteen working dogs presented to the Veterinary Specialist Group (VSG) and the Massey University Veterinary Teaching Hospital (MUVTH) with carpal injuries were prospectively treated using dorsal hybrid plating. Dogs were eligible if actively involved in farm, hunting or police work. Dogs had a standardised PCA surgical procedure performed, and similar instructions for post-operative care were provided. Dogs were re-evaluated clinically and radiographically at 6 weeks, 6 months, and 12 months after surgery. A questionnaire was completed by 12 owners, to assess each dog's working ability.

RESULTS: Twelve months following arthrodesis, 10/12 (83%) dogs could perform most or all duties normally. Eleven owners (92%) reported that the result of the surgery met their expectations, and nine owners (75%) were very satisfied with the outcome of the surgery. No owners were disappointed or very disappointed with the surgical outcome. Post-operative complications requiring surgical removal of the implant occurred in three (25%) dogs.

CONCLUSIONS: Unilateral PCA using a standardised surgical procedure and dorsal hybrid plating of carpal injuries has a good prognosis for working dogs in New Zealand to return to work.

CLINICAL RELEVANCE: These results may allow veterinarians to provide a more accurate prognosis to owners of working dogs that have debilitating carpal injury     

Multi-drug-resistant Enterococcus spp. as a cause of non-responsive septic synovitis in three horses

Abstract

CASE HISTORY: Three Thoroughbred horses, a 6-week-old filly (Case 1), a 15-year-old broodmare (Case 2) and a yearling filly (Case 3), sustained synovial sepsis secondary to trauma.

CLINICAL FINDINGS: Case 1 presented with a heel bulb laceration communicating with the distal interphalangeal joint. Arthroscopic lavage was performed and treatment commenced using systemic and local broad spectrum antimicrobial drugs. A pure growth of multi-drug-resistant (MDR) Enterococcus gallinarum was cultured from samples of synovium and joint fluid. Antimicrobial treatment was changed according to the susceptibility results. Response to treatment was poor and despite repeat arthroscopic lavage and intra-osseous regional perfusion of antimicrobials the filly was subject to euthanasia 24 days after the initial injury. Post-mortem examination confirmed septic synovitis, cartilage degeneration and osteomyelitis.

Case 2 sustained a full thickness wound to the carpus which was sharply debrided and closed. The wound dehisced with effusion within the tendon sheath. Drainage was established and treatment included systemic broad spectrum antimicrobials, topical lavage with povodine-iodine and manuka honey infusion. A mixed infection including MDR Enterococcus faecalis was cultured from the synovial fluid. Antebrachiocarpal joint effusion developed 21 days after initial injury and joint sepsis was confirmed. Arthroscopic lavage and tendon sheath debridement were performed, followed by treatment with systemic and local antimicrobials. The mare improved and was discharged. Three months later lameness recurred and corticosteroids were administered intra-articularly. The mare became non-weight bearing lame and was subject to euthanasia. Post-mortem examination confirmed joint sepsis of the antebrachiocarpal and intercarpal joint.

Case 3 presented with a complete articular open fracture of the tibial crest. Under general anaesthesia the fracture was stabilised and the wounds debrided and closed. Systemic broad-spectrum antimicrobials were administered. Six days later the wound dehisced and a bone fragment was removed. Three weeks post-surgery the wound deteriorated with a purulent discharge. Culture of the discharge revealed a mixed bacterial infection, including a MDR Enterococcus faecalis. Femoropatellar joint involvement was confirmed, and treatment included joint lavage, local and systemic antibiosis, and manuka honey instilled into the wound. The filly initially improved, and then deteriorated such that euthanasia was performed.

DIAGNOSIS: All three cases had synovial sepsis with MDR Enterococcus spp.

CLINICAL RELEVANCE: Increased awareness of MDR pathogens in equine wound infections is essential. Prompt diagnostic testing, appropriate therapy, infection control strategies and on-going monitoring and management are vital to limit the clinical impact of these organisms.     

Surgical management of suspected congenital luxation of the radial head in three dogs

Abstract

CASE HISTORIES: Three dogs, aged between 11 and 20 weeks, were presented with unilateral forelimb lameness, with an associated bony prominence on the lateral elbow.

CLINICAL FINDINGS AND DIAGNOSIS: Radiographs revealed a caudolateral luxation of the radial head in all cases, consistent with a diagnosis of suspected congenital luxation of the radial head. Surgical reduction and stabilisation involved open reduction of the radial head, and in two cases the use of a trans articular pin.

CLINICAL RELEVANCE: Congenital luxation of the radial head is an uncommon condition that has conflicting reports in the literature regarding its cause, heritability, breed predisposition, treatment and prognosis. The three cases here occurred in young dogs of English Bull Terrier, Jack Russell Terrier, and Staffordshire Bull Terrier breeds. Surgical reduction and stabilisation provided successful outcomes, in all cases. Only six cases of congenital luxation of the radial head managed surgically have previously been reported in the literature.     

Pathologic changes in extrahepatic organs and agglutinin response to Salmonella Gallinarum infection in Japanese quail fed Fusarium verticillioides culture material containing known levels of fumonisin B1

Three hundred 1-day-old Japanese quail (Coturnix coturnix japonica) were divided into two groups of 150 each. One group was maintained on quail mash alone, whereas Fusarium verticillioides culture material (FCM) was added to quail mash in the second group from 5 days of age and supplied 150 mg FB1/kg mash. At day 21, each group was further subdivided into two groups, yielding four groups with 75 birds apiece, which served as the control (group CX), the Salmonella Gallinarum alone group (group CS), the FB1 alone group (group FX), and the group fed FB1 and infected with Salmonella Gallinarum (group FS). An oral challenge with Salmonella Gallinarum organisms (2 x 10(4) colony-forming units [cfu]/ml) was given to groups CS and FS at 21 days of age. Three quail each, were necropsied on day 21 (0 day interval) from groups CX and FX, whereas at subsequent intervals, i.e., 1, 2, 3, 5, 7, 10, 14, and 21 days postinfection (DPI), they were sacrificed from all four groups (CX, CS, FX, and FS) to study the agglutinin response to Salmonella Gallinarum and pathologic changes. The agglutinin titers to Salmonella Gallinarum in the combination group (FS) were generally lower when compared with those in group CS. A reduction in the size of spleen along with depletion of white pulp, thinning of cardiomyocytes, lymphoid cell depletion from bursal follicles, and renal tubular nephrosis were characteristic pathologic changes in group FX. In contrast, there was mild to severe enlargement of spleen accompanied by necrosis and reticuloendothelial cell hyperplasia, pericarditis, myocarditis, and focal interstitial nephritis in groups CS. Similar but more severe lesions were observed in the combination group (FS). In addition, the flabby texture of heart, hydropericardium, and ascites were mainly observed in group FS. It is concluded that continuous presence of fumonisins at 150 mg/kg diet increases the severity of Salmonella Gallinarum infection in young Japanese quail.