Release of tumor necrosis factor-alpha from bovine alveolar macrophages stimulated with bovine respiratory viruses and bacterial endotoxins

The release of tumor necrosis factor-alpha (TNF-) from cultured bovine alveolar macrophages (BAM) was evaluated following stimulation of BAM with bovine herpesvirus-1 (BHV-1), parainfluenza-3 (PI-3) virus, bovine respiratory syncytial virus (BRSV), Escherichia coli 0111:B4 endotoxin, Pasteurella haemolytica type 1 endotoxin, Pasteurella multocida endotoxin, and virus/endotoxin combinations. A cytotoxic assay system using Georgia bovine kidney cells as targets was used to measure TNF- activity. The cytotoxic activity was neutralized by an anti-human TNF- monoclonal antibody.

Stimulation of BAM with 1 median tissue culture infectious dose (TCID₅₀) of live or ultraviolet (UV)-inactivated PI-3 virus/cell resulted in release of TNF- in significantly (P<0.05) higher amounts than sham-induced BAM. The quantities of TNF- released after live or UV-inactivated BHV-1 or BRSV induction were not significantly higher than sham-induced BAM. E. coli 0111:B4, P. haemolytica type 1 and P. multocida endotoxins stimulated TNF- release in a dose-dependent manner. Sequential exposure of BAM to 1 TCID₅₀ per cell of either live BHV-1, PI-3 virus or BRSV and then 5 μg ml⁻¹ of either E. coli 0111:B4, P. haemolytica type 1 or P. multocida endotoxin caused a significant (P<0.05) reduction in detectable TNF- in seven of nine virus/endotoxin combinations tested, when compared with 5 μg ml⁻¹ of endotoxin alone. Parainfluenza-3 virus/endotoxin combinations stimulated higher TNF- release when compared with other virus/endotoxin combinations. Five out of six test animals had serum-neutralizing antibodies to PI-3 virus, one out of six had serum-neutralizing antibodies to BHV-1, and two out of six had serum-neutralizing antibodies to BRSV, suggesting a possible relationship between serum neutralizing antibodies and TNF- release from in vitro cultivated BAM.     

Eastern equine encephalitis in a flock of emus (Dromaius novaehollandiae).

Eastern equine encephalitis (EEE) was diagnosed in a flock of emus in southeastern Louisiana. The outbreak involved juvenile and adult breeders ranging in age from 20 to 36 months, with an attack rate of 76% and a case fatality rate of 87%. The diagnosis was confirmed by isolation and characterization of the viral agent, and by detection of EEE antibody in two recovered emus. High mortality was preceded by marked depression, hemorrhagic diarrhea, and emesis of blood-stained ingesta. On postmortem examination, hemorrhagic enteritis and multiple petechia of viscera were observed. Microscopic changes included severe necrosis of hepatocytes, intestinal mucosa, and necrotizing vasculitis of the spleen and lamina propria of the intestine. No nervous system lesions were observed. This outbreak occurred concurrently with EEE in horses and was attributed to unseasonably heavy rainfall with an abundance of arthropod vectors and proximity to free-living reservoir host species.     

Chronic diarrhea associated with intestinal anomalies in a six-year-old dog.

A 6-year-old mixed-breed dog was referred because of chronic diarrhea, which had begun after antibiotic treatment for pulmonary disease. Intestinal bacterial overgrowth was presumptively diagnosed on the basis of intestinal mucosal cytologic examination and low serum cobalamin concentration. When the diarrhea did not respond to treatment, contrast radiography was performed and a short small intestine was found. The diarrhea could not be managed by medical treatment, and the dog was euthanatized. Necropsy revealed 2 blind intestinal pouches attached to a 46-cm length of small intestine connecting the stomach and colon. The dog's deteriorating condition was attributable to necrosis and rupture of 1 of the pouches.     

Cardiopathological observation on a case of persistent ventricular tachycardia in a pony mare.

A 20-year-old pony mare with persistent ventricular tachycardia (VT) was examined cardiopathologically. At necropsy, the heart was enlarged and rounded with both ventricular dilatation. In a longitudinal section of the ventricular septum, a large grayish white patchy lesion (5 x 25 mm) was detected in the relatively higher portion. Microscopically, the lesion was extensive myocardial fibrosis located in the vicinity of the proximal part of the left bundle branch. Partially the fibrotic lesion was in contact with the branch. Such a lesion might play an important role in creating a suitable background for the development of VT via automaticity or reentry mechanism.     

Effects of predator-induced visual and olfactory cues on 0+ perch (Perca fluviatilis L.) foraging behaviour

Abstract –  Foraging juvenile fish with relatively high food demands are usually vulnerable to various aquatic and avian predators. To compromise between foraging and antipredator activity, they need exact and reliable information about current predation risk. Among direct predator-induced cues, visual and olfactory signals are considered to be most important. Food intake rates and prey-size selectivity of laboratory-reared, naive young-of-the-year (YOY) perch, Perca fluviatilis , were studied in experiments with Daphnia magna of two size classes: 2.8 and 1.3 mm as prey and northern pike, Esox lucius , as predator. Neither total intake rate nor prey-size selectivity was modified by predator kairomones alone (water from an aquarium with a pike was pumped into the test aquaria) under daylight conditions. Visual presentation of pike reduced total food intake by perch. This effect was significantly more pronounced (synergistic) when visual and olfactory cues were presented simultaneously to foraging perch. Moreover, the combination of cues caused a significant shift in prey-size selection, expressed as a reduced proportion of large prey in the diet. Our observations demonstrate that predator-induced olfactory cues alone are less important modifiers of the feeding behaviour of naive YOY perch than visual cues under daylight conditions. However, pike odour acts as a modulatory stimulus enhancing the effects of visual cues, which trigger an innate response in perch.     

猪带绦虫六钩蚴cDNA文库的构建与筛选

在体外将猪带绦虫虫卵孵化为有活力的六钩蚴。采用商品化试剂盒抽提六钩蚴总 RNA、m RNA,反转录为双链c DNA,再加 Eco R Adapter。在去除小片段后 ,dsc DNA与 λgt11噬菌体 DNA连接 ,经蛋白包装 ,构建了猪带绦虫六钩蚴λgt11c DNA文库。该文库效价为 3.5× 10 9pfu/ m L ,重组 DNA片段大小为 0 .5～ 3.2 kb,平均为 1.6 kb,蓝白斑比 1∶ 9。用抗体探针对文库进行免疫学筛选 ,在消除非特异性反应的基础上筛选约 10 6 重组子 ,共得到 118株强阳性克隆 ;应用 PCR鉴定上述部分阳性克隆 ,均扩增到 0 .5 kb以上的片段。结果显示 ,构建的文库合格 ,含六钩蚴所有抗原基因 ,可用于六钩蚴 c DNA克隆的筛选 ;用免疫学与 PCR联合筛选 c DNA文库 ,可消除假阳性     

Spermatophore cryopreservation and artificial insemination of black tiger shrimp, Penaeus monodon (Fabricius)

To develop an appropriate cryopreservation protocol for spermatophores of black tiger shrimp, Penaeus monodon, three cryoprotectants (dimethyl sulphoxide (DMSO), methanol (MeOH) and ethylene glycol (EG)) at two concentrations (5% and 10%) were examined. Artificial implantation of spermatophores was also carried out to assess the fertilizing ability of fresh and post‐thaw spermatophores. Spermatophores were collected during consecutive regenerations (15‐day intervals) and assessed for qualitative and quantitative changes and also for fertilizing ability by implantation. The mean fertilization rate for artificial insemination using post‐thaw spermatophore was 79.9±3.7%, lower than the fertilization rates observed for artificial implantation using fresh spermatophore and natural mating. Mean hatch rates for fresh spermatophore, frozen‐thawed spermatophore and natural mating were 88.8±0.6%, 87.8±0.4% and 88.3±0.5%, respectively; and there was no difference among the three groups. The mean fertilization rate of spermatophores collected during the first stripping was higher (90.6±0.6) than during the second stripping (85.7±2.6), but the mean hatch rate was not different between the two strippings. The highest mean sperm viability (79.7±0.4%) was obtained from DMSO (5%), with no survival observed in the 10% MeOH treatment. Spermatophore weight, total sperm count and percentage of abnormal sperm were not different between spermatophores collected at the first and second stripping. This is the first study to report high fertilization and hatch rates from cryopreserved spermatophore using artificial implantation of spermatophore before spawning.     

Leishmaniasis phytotherapy. Nature's leadership against an ancient disease.

The use of phytotherapy to treat human diseases has its roots in pre-historical times. Despite the modern advances achieved in the field of synthetic chemistry, the most efficient drugs available have their genesis directly or indirectly related with the vegetal kingdom. Indigenous communities have long used plant extracts to treat illnesses. Many of these extracts have shown effective action, with new bioactive compounds being extracted and screened every year. These extracts have also proven to be good sources of therapeutic agents to the treatment of Leishmaniasis. This work highlights some of these agents, while trying to emphasize the importance of plants as a source of new and powerful drugs against this widespread disease.     

An evaluation of the pesera soil erosion model and its application to a case study in Zakynthos, Greece

Abstract. The Pan‐European Soil Erosion Risk Assessment (pesera ) model was evaluated using existing soil erosion data collected under various types of climate, vegetation, landscape and soil conditions. The data used represent a variety of typical Mediterranean land uses such as winter wheat, vines, olives and bare, stony land prevailing in hilly areas. Using this data, the model was calibrated for sediment transport by overland flow and results compared to measured soil erosion values from runoff plots and a watershed on a monthly basis. The performance of the model was assessed statistically, showing that it can be satisfactorily used for predicting soil erosion rates under the conditions included in the study. The overall model estimate including all the available experimental data was 0.69 t ha^?1 yr^?1 with a maximum error of 1.49 t ha^?1 yr^?1. After validation, the model was applied to a small watershed (60 ha) of great ecological importance for the sea turtle Caretta caretta. For this purpose, soil and vegetation maps were compiled from all the necessary data for applying the model. The model was run for three years using daily data from an existing nearby meteorological station. The predicted and measured soil erosion rates for a 7‐month period were 0.31 t and 0.18 t, respectively. Application of the model to each mapping unit showed the over‐riding importance of land use for sediment generation under the given climatic conditions. Bare land, occupying 5.5% of the watershed area, generated up to 69% of the total sediments estimated for the watershed. It is concluded that the pesera model can be used as a regional diagnostic tool under a range of soil, topographic and climatic conditions for identifying the best land use type and vegetation cover to protect hilly areas from soil erosion. The calculated overall root mean square error for the model is 0.06 t ha^?1 yr^?1, compared to a soil erosion rate of 0.04 t ha^?1 yr^?1, which can be tolerated for protecting the area for the sea turtle.     

Effects of low and high calcium intake prepartum on calcium mobilization rate around parturition in dairy cows

Forty-one dairy cows were fed a low (LCa-13 g/d) and a high (HCa-83.5 g/d) calcium ration in the 8 weeks prior to parturition and the effect on the Ca mobilization rate around parturition was studied. Plasma Ca values were stable in the LCa group around parturition. In the older cows of the HCa group a very slight decrease in the mean plasma Ca was observed: 2.58 mmol/l at 12-36 h ante partum decreased to 2.38 mmol/l at parturition. Hypocalcaemia, which commonly occurs around parturition, did not occur in 40 of the cows. A subclinical hypocalcaemia (1.8 mmol/l) occurred in one cow (parity 10) from the HCa group. To assess the efficiency of Ca mobilization, a severe hypocalcaemia (1.0 mmol/l) with clinical signs was induced by means of Na2EDTA infusion (0.90 mmol/min), starting at 10 h post-partum. The older cows in the LCa group required more Na2EDTA than those in the HCa group. Higher urinary hydroxyproline excretion in the week before parturition in the LCa than in the HCa group suggested a higher bone turnover. Plasma PTH levels around parturition were not significantly different between the groups. The amount of colostrum milked out in the first 10 h post-partum did not influence Ca homeostasis around parturition. The results contradict those of many other experiments in which hypocalcaemia was observed in cows ingesting high levels of Ca. It is concluded that the restricted feed intake prepartum possibly had a favourable effect on Ca homeostasis.