Alteration of extracellular collagen matrix in the myocardium of canines infected with Dirofilaria immitis.

The heart consists of cardiocytes and the interstitial extracellular matrix (ECM), which is made up mainly of collagens. The ECM has been suggested to be important in maintaining the structure and function of the heart. This investigation attempted to elucidate the changes in the ECM collagens in the hearts of canines with dirofilariasis. The ECM collagen fibrils of the heart are grouped into endomysial struts, epimysial weaves, and perimysial coils. In the present study, we used the modified silver impregnation technique to stain paraffin-embedded sections to demonstrate three types of ECM. The results revealed that the ECM content of the heart was significantly reduced in heartworm-infected dogs, and became fragmented and dissociated. In addition, the amounts of collagen in the septum (Sep), RVs and LVs in canines with dirofilariasis (Sep=11.55+/-0.65, RV=12.07+/-0.59, LV=11.72+/-0.62 microg/mg, n=24) were significantly lower (p<0.01) than that in the normal canines (Sep=15.09+/-0.72, RV=15.16+/-0.83, LV=14.91+/-0.89 microg/mg, n=8). These results indicated that heartworm infection induced the remodeling of the extracellular matrix, thus markedly altering the architecture and function of the heart.

Demonstration of vector competence of Culex quinquefasciatus (Diptera: Culicidae) for Setaria digitata.

In Taiwan, Setaria digitata infection causes a lumber paralysis in increasing number of cattle. Culex quinquefasciatus is one of the predominant mosquitoes, and it has been suspected that C. quinquefasciatus acts as a vector to Setaria nematodes prevalence but this was not confirmed. C. quinquefasciatus, Aedes albopictus and A. aegypti of various strains were investigated using an artificial infection system to evaluate their vector competence. After blood feeding at day 14, the number of larvae (stage III) per infected mosquito in A. aegypti (Liverpool strain), A. aegypti (Kaohsiung strain), A. aegypti (Tungan strain), C. quinquefasciatus (Taichung strain) and A. albopictus (Taichung strain) was 1.3 +/- 0.1, 1.3 +/- 0.1, 1.4 +/- 0.1, 1.0 +/- 0.0 and 0 +/- 0.0 (mean +/- S.E.M), respectively. The vector efficiency index of A. aegypti (Liverpool) was the highest among mosquitoes whereas A. albopictus showed a complete refractoriness to the infection. In conclusion, C. quinquefasciatus demonstrates its potential competence for serving as a transmission vector of S. digitata. This mosquito might therefore be responsible, at least in part, for the prevalence of cattle lumbar paralysis in Taiwan. This is the first report of C. quinquefasciatu demonstrating its vector competence for S. digitata.

The effect of different extenders on post-thaw sperm survival, acrosomal integrity and longevity in cryopreserved semen of Formosan Sika deer and Formosan Sambar deer.

This study investigates the efficacy of five extenders in contributing to the outcome of semen cryopreservation in Formosan Sika and Sambar deer. Pooled semen (n=4) of six males of each breed was used. In Sika deer, semen collection rate was 96% (23/24) over all electro-ejaculations. Volume, sperm motility and sperm concentration of fresh ejaculates was 0.5+/-0.4 ml, 77+/-6% and 1471.3+/-940.0 x 10(6) ml(-1), respectively. Post-thaw motility in respective extender was A: 66+/-16%; B: 71+/-2%; C: 73+/-6%; D: 9+/-4% and E: 26+/-12% (mean+/-S.D.). In extender C (74+/-14%) more viable spermatozoa were preserved than in the others (A: 64+/-10%; B: 48+/-11%; D: 41+/-16%; E: 47+/-6%; P<0.05). Acrosomal integrity was not influenced by extender composition. Post-thaw motility did not decrease during a 4-h incubation period, irrespective of the extender used (P>0.05). In Sambar deer, semen collection rate was 88% (21/24) over all electro-ejaculations. Volume, sperm motility and sperm concentration of fresh ejaculates was 1.3+/-0.5 ml, 82+/-4% and 379.1+/-252.2 x 10(6) ml(-1), respectively. Post-thaw motility was in respective extenders A: 69+/-2%; B: 74+/-6%; C: 73+/-2%; D: 13+/-6% and E: 31+/-20%. Extenders B and C were superior (P>0.05) with respect to sperm motility. Similarly, post-thaw viability in extenders A (70+/-7%), B (76+/-7%) and C (79+/-2%) was higher than that D (25+/-19%) and E (29+/-17%) (P<0.01). Sperm acrosomal integrity was better preserved in extenders B (86+/-4%) and C (83+/-4%) than in extenders A (54+/-13%), D (39+/-22%) and E (46+/-22%) (P<0.05). Post-thaw sperm longevity in extender A reduced from 69 to 16% during incubation (P<0.05) whereas only a slight decrease was observed in the other extenders after 4 h. In conclusion these data show that egg-yolk-Tris-Tes-glycerol based extender C containing Equex STM paste is optimal for freezing semen of Formosan Sika deer while egg-yolk-Tris-citric acid-glycerol based extender B containing Equex and extender C are superior in semen cryopreservation to others for Formosan Sambar deer.

Effect of thuringiensin on adenylate cyclase in rat cerebral cortex.

The purpose of this work is to evaluate the effect of thuringiensin on the adenylate cyclase activity in rat cerebral cortex. The cyclic adenosine 3'5'-monophosphate (cAMP) levels were shown to be dose-dependently elevated 17-450% or 54-377% by thuringiensin at concentrations of 10 microM-100 mM or 0.5-4 mM, due to the activation of basal adenylate cyclase activity of rat cerebral cortical membrane preparation. Thuringiensin also activated basal activity of a commercial adenylate cyclase from Escherichia coli. However, the forskolin-stimulated adenylate cyclase activity in rat cerebral cortex was inhibited by thuringiensin at concentrations of 1-100 microM, thus cAMP production decreased. Furthermore, thuringiensin or adenylate cyclase inhibitor (MDL-12330A) reduced the forskolin (10 microM)-stimulated adenylate cyclase activity at concentrations of 10 microM, 49% or 43% inhibition, respectively. In conclusion, this study demonstrated that thuringiensin could activate basal adenylate cyclase activity and increase cAMP concentrations in rat cerebral cortex or in a commercial adenylate cyclase. Comparing the dose-dependent effects of thuringiensin on the basal and forskolin-stimulated adenylate cyclase activity, thuringiensin can be regarded as a weak activator of adenylate cyclase or an inhibitor of forskolin-stimulated adenylate cyclase.

Cerebrospinal setariosis with Setaria marshalli and Setaria digitata infection in cattle.

Setaria digitata and S. marshalli larvae were observed in the cerebrospinal cavity of 2 paralyzed cattle in Taiwan. The 2 affected cattle showed quadriplegia and lumbar paralysis, respectively. At necropsy, which was performed 7 days after the 7-month-old cattle became quadriplegic, three and nineteen S. marshalli larvae as well as two female adult worms were found in the cranial cavity, spinal cavity and peritoneal cavity of the cattle, respectively. Necropsy on the other 8-month-old cattle was also performed 3 days after it showed lumbar paralysis, and ten S. digitata larvae were found in the spinal cavity. In both cattle, many mononuclear inflammatory cells mixed with a few eosinophils were seen accumulated in the connective tissue around the root of the spinal nerves. Infiltration of eosinophils and mononuclear inflammatory cells into the epidura and arachnoidea of the brain were also observed. The major inflammatory cell was lymphocytes, but neutrophils and eosinophils were also present. The number of cells in the cerebrospinal fluid collected initially from the two affected cattle were 105/0.01 ml and 143/ 0.01 ml, respectively. This is the first report of cerebrospinal setariosis in cattle associated with S. marshalli.

Pulmonary toxicity of thuringiensin administered intratracheally in Sprague-Dawley rats.

The purpose of this work is to evaluate the pulmonary toxicity of purified thuringiensin in Sprague-Dawley rats. Rats were intratracheally instillated with 0, 0.4, 0.8, 1.6, 3.2, 6.4 and 9.6 mg/kg body weight of thuringiensin. The results indicated that the acute pulmonary LD(50) of thuringiensin for rats was 4.4 mg/kg. The total number of inflammatory cells and lactate dehydrogenase (LDH) activity in the bronchoalveolar lavage (BAL) fluid increased in a dose-dependent manner after thuringiensin instillation. Furthermore, an effective dose of 1.6 mg/kg was selected for the time course study of pulmonary toxicity. The treated animals showed a significant increase in the weights of the lungs, hydroxyproline levels in the lungs and total number of cells in BAL fluid 2, 4, 7, 14, 28 and 56 days after treatment. In comparison with the control, the total protein concentrations in BAL fluid were increased by 361, 615, 116, 41, 34 and 41%, after 2, 4, 7, 14, 28 and 56 days, respectively. The LDH activity in BAL fluid showed a significant increase after 1, 2, 4, 7, 14, 28 and 56 days. The increases in fibronectin levels were 164, 552, 490, 769, 335, 257 and 61% at the corresponding times, but neither tumor necrosis factor nor interleukin-1 increased. The treated rats presented abnormal histology including distributed inflammation in the bronchioles and alveoli, bronchial cellular necrosis on days 1 and 2, and areas of septal thickening with cellular infiltration and collagen deposit in the intestinal and alveolar spaces on days 4-56. Based on these biochemical and pathological parameters, intratracheal instillation of purified thuringiensin might cause significant pulmonary toxicity in rats.

Effect of ozone treatment on Eimeria colchici oocysts.

The effect of ozone on the inhibition of the sporulation of Eimeria colchici oocysts in vitro was examined. Lower sporulation ratios were found to correspond directly to longer ozone exposure time. In pheasants, Phasianus colchicus, orally inoculated with ozone-treated oocysts, lower mortality and lower oocysts per gram of feces were observed as compared with birds given untreated oocysts. Thus, treatment of E. colchici oocysts with ozone alone was observed to partially inhibit the growth and infectivity of the oocysts.