Use of an antineoepitope antibody for identification of type-II collagen degradation in equine articular cartilage.

OBJECTIVE: To develop an antibody that specifically recognizes collagenase-cleaved type-II collagen in equine articular cartilage. SAMPLE POPULATION: Cartilage specimens from horses euthanatized for problems unrelated to the musculoskeletal system. PROCEDURE: A peptide was synthesized representing the carboxy- (C-) terminus (neoepitope) of the equine type-II collagen fragment created by mammalian collagenases. This peptide was used to produce a polyclonal antibody, characterized by western analysis for reactivity to native and collagenase-cleaved equine collagens. The antibody was evaluated as an antineoepitope antibody by ELISA, using peptides +/- an amino acid at the C-terminus of the immunizing peptide. Collagen cleavage was assayed from equine articular cartilage cultured with interleukin-1 (IL-1), +/- a synthetic MMP inhibitor, BAY 12-9566. Cartilage specimens from osteoarthritic and nonarthritic joints were compared for antibody staining. RESULTS: An antibody, 234CEQ, recognized only collagenase-generated 3/4-length fragments of equine type-II collagen. This was a true antineoepitope antibody, as altering the C-terminus of the immunizing peptide significantly decreased competition for binding in an inhibition ELISA. The IL-1-induced release of type-II collagen fragments from articular cartilage was prevented with the MMP inhibitor. Cartilage from an osteoarthritic joint of a horse had increased staining with the 234CEQ antibody, compared with normal articular cartilage. CONCLUSIONS AND CLINICAL RELEVANCE: We generated an antineoepitope antibody recognizing collagenase-cleaved type-II collagen of horses. This antibody detects increases in type-II collagen cleavage in diseased equine articular cartilage. The 234CEQ antibody has the potential to aid in the early diagnosis of arthritis and to monitor treatment responses.     

Serum lipase determination in the dog: a comparison of a titrimetric method with an automated kinetic method

An enzymatic, kinetic method for determining serum lipase activity was evaluated and compared to a standard manual method for use in dogs. The kinetic method was a commercial kit adapted for use on a tandem access clinical chemistry analyzer and utilized a series of coupled enzymatic reactions based on the hydrolysis of 1,2-diglyceride by lipase. The manual method was the Cherry-Crandall technique based on the titration of base against the acid formed by hydrolysis of an olive oil substrate by lipase. The correlation between the two methods was very good (r = 0.94). The reference range for 56 clinically healthy dogs assayed by the kinetic method was 90 to 527 U/L. Diseases associated with a greater than twofold elevation in serum lipase activity as determined by the kinetic method included pancreatitis, gastritis with liver disease, and oliguric renal failure with metabolic acidosis. In some cases, pancreatitis was seen with other clinical problems, such as gastroenteritis, diabetic ketoacidosis, duodenal mass, disseminated intravascular coagulation, and septic peritonitis. Diseases associated with serum lipase activity within the reference range or elevated less than twofold included gastritis, gastric ulcer, cholestasis, phenobarbital-induced hepatopathy, colitis, copper hepatopathy, abdominal hematoma, apocrine gland adenocarcinoma, and thrombocytopenia with pneumonia.     

The intensity of resistance by mature Merino ewes against Haemonchus contortus and Tichostrongylus colubriformis in single-species and combined-species infection

Three-year-old, non-lactating and non-pregnant Merino ewes, raised on pasture under a program of strategic treatment with anthelmintic and found to be extremely resistant to "trickle" infection with Haemonchus contortus, were given single-dose infections with either H. contortus or Trichostrongylus colubriformis or both species together. The purpose was to ascertain the intensity of protective immunity against the 2 parasites in sheep with immunity acquired from a presumably slight exposure to infection. To provide a criterion, some infected ewes were immunosuppressed with corticosteroid, dexamethasone. Untreated ewes were extremely resistant to challenge infection with either 15,000 or 150,000 H. contortus or 15,000 T. colubriformis. Surprisingly, when mixed infection was given, egg counts for H. contortus were significantly elevated compared with infection by that species alone. Antibody to antigens from infective larval and adult H. contortus was measured in serum by enzyme-linked-immunosorbent assay (ELISA) during the course of infection. Serum titres against larval antigens were significantly depressed when infections with either H. contortus or T. colubriformis were permitted by immunosuppression with dexamethasone, whereas those against adult antigen were depressed when infection with T. colubriformis was permitted.     

The effects of duration and concentration of plasma progesterone on the fertility of post-partum cows treated with pregnant mare serum gonadotrophin and intravaginal progesterone

The effects of duration of insertion of progesterone releasing devices (PRIDs) on the concentrations of plasma progesterone (plasma P4) and the relationship of these with fertility were examined respectively in ovariectomised and intact Bos taurus cattle. In experiment 1, ovariectomised cows (n = 14) were treated with PRIDs to produce high and low plasma P4 for 7 or 14 days giving 4 treatment groups designated H7, H14, L7 or L14. Mean plasma P4 in the H14 and L14 groups were 6.6 and 6.0 ng/ml respectively, during the first week and 5.4 and 3.8 ng/ml respectively, during the second. Mean plasma P4 of 6.3 and 4.2 ng/ml were observed in H7 and L7. In experiment 2, suckled cows (n = 171) received the same PRID treatments as cows in experiment 1, plus either 375 or 750 iu pregnant mare serum gonadotrophin, and subsequent calving rates were recorded. These were significantly lower after L7 than the other treatments (12% v 38%, P less than 0.025). These results showed that fertility after oestrus was closely related to plasma P4 observed before oestrus and indicated that treatment with progesterone for as little as 1 week can produce normal fertility providing plasma P4 attain a critical threshold during treatment.     

Population dynamics and interactions between endemic entomopathogenic nematodes and annual bluegrass weevil populations in golf course turfgrass

Entomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) (EPNs) are generalist obligate pathogens present in the soil of most ecosystems. They have the potential to infect a broad host range, yet the potential for endemic EPNs to regulate soil-dwelling insect populations has received limited attention. We investigated the population dynamics of endemic EPNs to determine their ability to regulate annual bluegrass weevil (Listronotus maculicollis) (Coleoptera: Curculionidae) populations, a major pest of turfgrass in the northeastern United States. Weekly sampling of nematode and L. maculicollis populations was conducted in untreated fairway transects on three golf courses in New Jersey between April and October of 2006 and 2007. Steinernema carpocapsae and Heterorhabditis bacteriophora were found infecting all weevil stages from third instar to teneral adults. Both EPN species exhibited a distinct seasonality, appearing in high densities in the weeks immediately following high densities of first generation weevils in the soil. A positive temporal relationship was observed between densities of EPN-infected weevils and weevil stages between third instar and pupa, but L. maculicollis generational mortality due to EPN infection was highly variable between years and sites, ranging between 0 and 50%. Although infection densities and larval densities were positively correlated, per capita mortality did not increase with increased weevil densities. EPN distribution dynamically cycled between aggregated and uniform throughout the season across fairways. Few significant relationships were found to support the hypothesis that weevil spatial dispersion influences EPN spatial dispersion. The variability in EPN seasonal occurrence and generational impact on L. maculicollis together with the lack of spatial association with L. maculicollis suggest an inability of endemic EPN populations to reliably regulate weevil populations on golf course fairways.     

Effects of suckling on oestradiol benzoate induced LH release in post-partum, ovariectomised cows

Twelve suckling and 12 non-lactating post-partum cows were treated with a progesterone-releasing pessary for 7 or 14 d followed by an injection of 500 micrograms oestradiol benzoate (ODB) 24 h after pessary removal or no injection. Suckling had no significant effect on plasma progesterone concentrations (plasma P4) or plasma luteinising hormone concentrations (plasma LH) during pessary insertion. After pessary removal plasma LH rose in response to ODB. The extent of the rise was similar in suckling and non-lactating cows treated with progesterone for 14 d but was significantly lower in non-lactating than suckling cows (P less than 0.025) treated for 7 d. These findings indicate that suckling increases the responsiveness of plasma LH in post-partum cows following progesterone and oestrogen treatment compared to non-lactating cows and that extended treatment with progesterone can remove this difference.     

Animal health monitoring and surveillance in Switzerland

Switzerland has traditionally used a passive disease reporting system for all notifiable diseases. This type of system is not suitable for the documentation of very low prevalences (freedom from disease), sub-clinical cases and non-notifiable diseases. In order to meet the high international standards for animal health surveillance and to fulfil the general need for sound animal health data, Switzerland has evaluated the feasibility of modern monitoring and surveillance concepts. In general, the principle of active surveillance has been acquired and is now being applied whenever possible. In this paper, several examples of Swiss surveillance systems are presented and discussed. They include systematic testing of random population samples, carcase screening at abattoirs and sentinel herd monitoring.     

Comparative Earth History and Late Permian Mass Extinction

The repeated association during the late Neoproterozoic Era of large carbon-isotopic excursions, continental glaciation, and stratigraphically anomalous carbonate precipitation provides a framework for interpreting the reprise of these conditions on the Late Permian Earth. A paleoceanographic model that was developed to explain these stratigraphically linked phenomena suggests that the overturn of anoxic deep oceans during the Late Permian introduced high concentrations of carbon dioxide into surficial environments. The predicted physiological and climatic consequences for marine and terrestrial organisms are in good accord with the observed timing and selectivity of Late Permian mass extinction.     

Adult neural function requires MeCP2

Rett syndrome (RTT) is a postnatal neurological disorder caused by mutations in MECP2, encoding the epigenetic regulator methyl-CpG-binding protein 2 (MeCP2). The onset of RTT symptoms during early life together with findings suggesting neurodevelopmental abnormalities in RTT and mouse models of RTT raised the question of whether maintaining MeCP2 function exclusively during early life might protect against disease. We show by using an inducible model of RTT that deletion of Mecp2 in adult mice recapitulates the germline knock-out phenotype, underscoring the ongoing role of MeCP2 in adult neurological function. Moreover, unlike the effects of other epigenetic instructions programmed during early life, the effects of early MeCP2 function are lost soon after its deletion. These findings suggest that therapies for RTT must be maintained throughout life.