Evaluation of various compounds to inhibit activity of matrix metalloproteinases in the tear film of horses with ulcerative keratitis

OBJECTIVE: To examine in vitro effects of various antiproteolytic compounds on activity of matrix metalloproteinase (MMP)-2 and -9 in the tear film of horses with active corneal ulcers. SAMPLE POPULATION: Samples of tear film obtained from the eyes of 34 horses with active ulcerative keratitis. PROCEDURE: Horses were sedated, and tear samples were collected from the lower fornix of 34 ulcerated eyes by use of capillary tubes. The protease inhibitors 0.2% EDTA, 0.1% doxycycline, 10% N-acetylcysteine (NAC), 0.1% solution of a modified dipeptide that contains hydroxamic acid (ie, ilomostat), 0.1% alpha1-proteinase inhibitor (PI), 0.5% alpha1-PI, and 100% fresh equine serum (ES) were used to treat pooled samples. Amount of latent and active MMP-2 and -9 was measured by optical density scanning of gelatin zymograms of treated and untreated tear samples. RESULTS: Pooled tear samples obtained from ulcerated eyes contained the latent and active forms of MMP-2 and -9. Compared with MMP activity in untreated samples, total MMP activity (sum of all bands detected) observed on the gelatin zymogram gels was reduced by 99.4% by EDTA, 96.3% by doxycycline, 98.8% by NAC, 98.9% by ilomostat, 52.4% by 0.1% alpha1-PI, 93.6% by 0.5% alpha1-PI, and 90.0% by ES. CONCLUSIONS AND CLINICAL RELEVANCE: We documented that EDTA, doxycycline, NAC, ilomostat, alpha1PI, and ES inhibited MMP activity in vitro. Because these compounds use different mechanisms to inhibit various families of proteases in the tear film of horses, a combination of these protease inhibitors may be beneficial for treatment of corneal ulcers in horses.     

Evaluation of survival of Actinobacillus pleuropneumoniae and Haemophilus parasuis in four liquid media and two swab specimen transport systems

OBJECTIVE: To determine duration and rates of recovery of Actinobacillus pleuropneumoniae and Haemophilus parasuis from 4 liquid media and 2 swab specimen transport systems and compare findings with those of Escherichia coli. SAMPLE POPULATION: One strain each of A pleuropneumoniae (biovar 1, serotype 1), H parasuis (serovar 5), and E coli (serotype O149:K91:H19). PROCEDURE: Strains were incubated in brain heart infusion broth supplemented with horse serum and other nutrients or in horse serum alone, with and without nicotinamide-adenine dinucleotide in both instances, for 150 days at 4 degrees C or room temperature (21 degrees C). Similarly, strains were tested in Stuart and Amies transport systems after storage at room temperature for 8 days. RESULTS: Colony counts greater than those of the initial inoculum were observed after incubation in horse serum for A pleuropneumoniae but not for H parasuis. Overall, incubation at 4 degrees C in the 4 liquid media resulted in longer recovery duration and higher rates than at room temperature. Culture of H parasuis resulted in lower recovery rates and shorter durations of recovery than culture of A pleuropneumoniae, except for culture in horse serum. Haemophilus parasuis survived longer than A pleuropneumoniae in the transport systems, and all organisms survived longer in the Amies system. CONCLUSIONS AND CLINICAL RELEVANCE: Survival of A pleuropneumoniae and H parasuis indicated that horse serum prolongs survivability, which may result in exposure of more animals during a prolonged period. The Amies system might be a good choice for collection of clinical samples from animals, especially for recovery of H parasuis.     

Evaluation of the safety of ivermectin-praziquantel administered orally to pregnant mares

OBJECTIVE: To evaluate the safety of an orally administered ivermectin and praziquantel paste with regard to variables associated with clinical findings, parturition, lactation, maternal care, and neonate viability in pregnant mares. ANIMALS: 40 pregnant mares. PROCEDURE: Mares were randomly allocated into treatment (n = 20) and control (20) groups and administered a placebo or 3 times the therapeutic dosage of ivermectin (0.6 mg/kg) and praziquantel (4.5 mg/kg) at 14-day intervals until parturition. Physical examinations were performed on mares and their foals after parturition (on postpartum days 30, 60, and 90) to identify any drug-related effects. As an aid in assessing general health, hematologic and serum biochemical analyses were performed monthly on the mares. RESULTS: In blood constituents, minor alterations that were not biologically important were observed. Reproductive performance was not affected by the unusual treatment duration or high dosage, although the drugs were administered during a crucial period of equine embryonic development (30 to 60 days). Neither adverse effects on mares nor abortions occurred. Follow-up evaluations of the foals for a 3-month period did not detect any abnormalities. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of the ivermectin-praziquantel paste appears to be safe in pregnant mares and their foals.     

The effects of GnRH and adrenergic agents on PRL and beta-endorphin secretion by porcine pituitary cells in vitro

The direct effects of alpha- and beta-adrenergic agents on PRL and beta-endorphin (beta-END) secretion in vitro by porcine pituitary cells have been investigated. Pituitary glands were obtained from mature gilts, which were ovariectomised (OVX) one month before slaughter. Ovariectomised gilts, assigned to four groups, were primed with: (1) vehicle (OVX); (2) and (3) oestradiol benzoate (EB; 2.5 mg/100 kg b.w.) at 30-36 h (OVX+EB I) and 60-66 h (OVX+EB II) before slaughter, respectively; and (4) progesterone (P4; 120 mg/100 kg b.w.) for 5 consecutive days before slaughter (OVX+P4). Isolated anterior pituitary cells were submitted to 3.5 h incubation in the presence of GnRH, alpha- and beta-adrenergic agonists [phenylephrine (PHEN) and isoproterenol (ISOP), respectively], or alpha- and beta-adrenergic blockers [phentolamine (PHENT) and propranolol (PROP), respectively]. The culture media were assayed for PRL (exp. I) and beta-endorphin-like immunoreactivity (beta-END-LI) (experiment II). In experiment I, GnRH did not influence PRL release by pituitary cells in all experimental groups. Some of tested doses of adrenergic agonists, PHEN and ISOP, increased PRL release from pituitary cells of OVX gilts, but not from those of OVX+EB I animals. In the OVX+EB II group, PHEN alone, but ISOP with PROP, potentiated PRL secretion by the cells. In OVX+P4 animals, PHEN alone or in combination with PHENT and also ISOP alone or with PROP enhanced PRL output from the cells. In experiment II, addition of GnRH increased beta-END-LI release from pituitary cells only in the OVX+EB II group. PHEN and PHENT potentiated beta-END-LI secretion by pituitary cells in OVX+EB II and OVX+P4 groups, while ISOP and PROP increased beta-END-LI secretion by the cells of OVX and OVX+EB II animals. In turn, in the OVX+EB I group, effect of PHENT and PROP on PRL secretion by pituitary cells was inhibitory. In conclusion, our results suggest that adrenergic agents can modulate PRL and beta-END secretion by porcine pituitary cells in a manner dependent on the hormonal status of gilts.     

Efficacy of formic acid in gel for Varroa control in Apis mellifera L.: importance of the dispenser position inside the hive

The efficacy of formic acid in a gel matrix was evaluated in two groups of honeybee colonies. In Group 1, a dispenser with 120 g of formic acid (70%) in gel was placed on the brood combs and another dispenser with the same dose was located on the hive bottom (total dose, 240 g). Group 2 received two doses of 240 g of formic acid (70%) in gel and each application was applied in two dispensers containing 120 g of the formic acid solution each and they were located over the brood chamber (total dose, 480 g). In Group 2, the period between both applications was 15 days, and the efficacies after the first and both applications were calculated. Significant differences were registered for final efficacy between both groups. When final efficacy of Group 1 was compared with efficacy after first application of Group 2, significant differences were found (P=0.0005). Same doses in different positions within the hive have different final efficacy. The higher efficacy was registered when the dispensers were placed over brood combs and on the hive bottom. It is suggested that efficacy is related to dispenser position within the hive.     

Predominance of the papGII allele with high sequence homology to that of human isolates among avian pathogenic Escherichia coli (APEC)

Avian pathogenic Escherichia coli (APEC) are often found in poultry and are responsible for a set of diseases, commonly referred to as avian colibacillosis. One of the important virulence factors is adhesion to different epithelial surfaces, which is mediated by pili. P pili are thought to play a role by means of their PapG adhesin, which occurs in three molecular variants: PapGI, PapGII and PapGIII. This study is the first to determine and analyse the distribution of the different papG alleles in APEC. Our results show a significant predominance of the papGII allele above all other alleles or allele combinations. No statistically significant associations could be found between papG allele distribution and the type of bird, organ of isolation and O serogroup. Finally, the papGII and papGIII sequences showed high homology with mammalian (including human) source papG sequences.     

Dynamics of porcine circovirus type 2 infection in a herd of pigs with postweaning multisystemic wasting syndrome

OBJECTIVE: To determine the pattern of infection for porcine circovirus type 2 (PCV2) in a herd of pigs with postweaning multisystemic wasting syndrome (PMWS). ANIMALS: 29 sows and 250 pigs. PROCEDURE: Blood samples were collected from all 3-, 7-, and 12-week old pigs and 59 pigs at 28 weeks of age. Pigs that died during the study were necropsied. Porcine parvovirus and PCV2 antibodies were assayed. A polymerase chain reaction (PCR) was used to detect PCV2 genome in serum of selected pigs. RESULTS: The PMWS started when pigs were 8 weeks old, with a prevalence of 30% in 8- to 10-week-old pigs. Eighty-three pigs died during the period between 3 and 12 weeks of age. Microscopic lesions consistent with PMWS were observed, and PCV2 nucleic acid was detected (50 of 68 pigs). Antibodies to PCV2 decreased from 3 to 7 weeks of age, increased at 12 weeks of age, and were maintained until 28 weeks of age. One sow had a positive result for PCR of serum. Nine, 37 and 8 pigs had PCV2 genome in serum obtained at 7, 12, and 28 weeks of age, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Infection with PCV2 coincided with severe clinical signs; however, infected 28-week-old pigs did not have evidence of disease. Immunity declined over time in young pigs. A long duration of PCV2 viremia was apparent in a high percentage of infected pigs, which may affect transmission and persistence of the virus in a herd.     

Facet joint geometry and intervertebral disk degeneration in the L5-S1 region of the vertebral column in German Shepherd dogs

OBJECTIVE: To evaluate the possible association between facet joint geometry and intervertebral disk degeneration in German Shepherd Dogs. ANIMALS: 25 German Shepherd Dogs and 11 control dogs of similar body weight and condition. PROCEDURE: Facet joint angles in the caudal portion of the lumbar region of the vertebral column (L5-S1) were measured by use of computed tomography, and the intervertebral discs were evaluated microscopically. The relationship between facet joint geometry and disk degeneration was evaluated by use of statistical methods. RESULTS: German Shepherd Dogs had significantly more facet joint tropism than control dogs, but an association with disk degeneration was not found. However, German Shepherd Dogs had a different facet joint conformation, with more sagittally oriented facet joints at L5-L6 and L6-L7 and a larger angle difference between the lumbar and lumbosacral facet joints, compared with control dogs. CONCLUSIONS AND CLINICAL RELEVANCE: A large difference between facet joint angles at L6-L7 and L7-S1 in German Shepherd Dogs may be associated with the frequent occurrence of lumbosacral disk degeneration in this breed.