草鱼NK-lysin基因的克隆、原核表达与活性分析

为研究草鱼抗菌肽NK-lysin基因的组成结构、在组织中的表达差异及其抑菌活性,实验根据已知鱼类NK-lysin基因保守区序列设计上下游引物,采用RT-PCR和RACEPCR技术克隆草鱼NK-lysin基因(Cinkl),RT-PCR进行各组织间的表达分析,并构建Cinkl成熟肽的原核表达载体,采用琼脂糖孔穴扩散法检测重组蛋白的抑菌活性。结果显示,Cinkl c DNA全长768 bp,编码121个氨基酸;基因组DNA全长3 361 bp,包含4个外显子和3个内含子。经多序列比对分析和结构域预测表明,Ci Nkl氨基酸序列具有SAPLIP家族的特征:含有6个保守的半胱氨酸和saposin B蛋白结构域,与斑马鱼Nklc、Nkld的相似性分别为57.98%和63.03%。构建NK-lysin氨基酸序列系统进化树,发现Ci Nkl与斑马鱼Nklc、Nkld聚为一支。RT-PCR检测结果显示,Cinkl m RNA在所有检测的草鱼组织中均有表达,在脾脏中表达量最高,心脏、鳃、中肾和头肾的表达量次之,皮肤、脑、肝脏和肠有微量表达。p ET-28b-MBP与Ci Nkl成熟肽序列构建的原核表达质粒在宿主菌Rosetta(DE3)表达出约57 ku的可溶性融合蛋白,利用Ni-NTA His·Bind树脂进行纯化,Ci Nkl重组蛋白对G–细菌如大肠杆菌M15株、嗜水气单胞菌,以及G+细菌如金黄色葡萄球菌均具有抑菌活性。基于草鱼NK-lysin的组织表达特征和重组蛋白的抑菌活性,推测NK-lysin在草鱼的先天性免疫防御中发挥调节作用。     

Testing newly introduced ornamental plants to the arid climate of Kuwait

Thirty five ornamental plants were introduced into the Ahmadi Bioremediated Soil Park to screen and closely monitor the growth performance of these new introductions under the local environmental conditions. Out of these 35 plant species, 15 were planted in both bioremediated and agricultural soils to assess their suitability for growing in bioremediated soils. Data on vegetative growth was recorded to assess the performance of newly-introduced plants, whereas, the heat stress was documented of Bauhinia blakeana, where no visible effects of petroleum pollutants on plant growth were observed. Performance of each plant group is discussed separately in the text. Overall, plant survival was 72.73% for all plant groups. Generally, plants that failed to grow in bioremediated soil also did not survive in the agricultural soil, indicating that they were sensitive to heat stress more than the presence of petroleum contaminants in the soil. New plant species that were able to establish and perform satisfactorily will be used in greenery projects after they are fully naturalized to the prevailing climatic conditions.     

Surgical management of an abdominal abscess in a Malayan tapir

A captive Malayan tapir was observed to have inappetence, weight loss, signs of depression, mild dehydration and diar rhoea. Haematological and serum biochemical tests showed anaemia, hypoproteinaemia, hyperfibrinogenaemia and neu-trophilia with a left shift. Ultrasonic examination of the abdomen under anaesthesia revealed a well-encapsulated abscess. The abscess was marsupialised to the ventral body wall. Culture of the pus produced a mixed bacterial growth. Antimicrobial therapy was based on bacterial sensitivity results. Follow-up ultrasonic examinations showed resolution of the abscess. Ninety-one days after surgery the tapir began regurgitating food and water. An abscess originating from the stomach and occluding the lumen of the duodenum was identified at surgery. The abscess ruptured during surgical manipulations and the tapir was euthanased.     

Sperm Morphology Assessment in Captive Neotropical Primates

The main objective of this study was to evaluate sperm morphology in four neotropical primate species to compare the sperm morphological traits and the sperm morphometric parameters as a basis for establishing normative sperm standards for each species. Data from 80 ejaculates collected from four primate species, Callithrix jacchus, Callimico goeldii, Alouatta caraya and Ateles geoffroyi, were analysed for detection of sperm morphological alterations using subjective World Health Organization (WHO‐2010) standards and Sperm Deformity Index (SDI) criteria, objective computer‐assisted sperm morphometry analysis (CASMA) and subpopulation sperm determination (SSD) methods. There were multiple differences (p < 0.01) observed among primate species in values obtained from WHO‐2010, SDI, CASMA and SSD sperm analysis methods. In addition, multiple significant positive and negative correlations were observed between the sperm morphological traits (SDI, Sperm Deformity Index Head Defects, Sperm Deformity Index Midpiece Defects, Sperm Deformity Index Tail Defects, Normal Sperm, Head Defects, Midpiece Defects and Tail Defects) and the sperm morphometric parameters (SSD, Area (A), Perimeter (P), Length (L), Width (W), Ellipticity, Elongation and Rugosity) (p ≤ 0.046). In conclusion, our findings using different evaluation methods indicate that pronounced sperm morphological variation exists among these four neotropical primate species. Because of the strong relationship observed among morphological and morphometric parameters, these results suggest that application of objective analysis methods could substantially improve the reliability of comparative studies and help to establish valid normative sperm values for neotropical primates.     

Evaluation of chemical restraint,isoflurane anesthesia and methadone or tramadol as preventive analgesia in spotted pacas (Cuniculus paca) subjected to laparoscopy

ObjectiveTo evaluate the efficacy and cardiopulmonary effects of ketamine–midazolam for chemical restraint, isoflurane anesthesia and tramadol or methadone as preventive analgesia in spotted pacas subjected to laparoscopy.Study designProspective placebo-controlled blinded trial.AnimalsA total of eight captive female Cuniculus paca weighing 9.3 ± 0.9 kg.MethodsAnimals were anesthetized on three occasions with 15 day intervals. Manually restrained animals were administered midazolam (0.5 mg kg^–1) and ketamine (25 mg kg^–1) intramuscularly. Anesthesia was induced and maintained with isoflurane 30 minutes later. Tramadol (5 mg kg^–1), methadone (0.5 mg kg^–1) or saline (0.05 mL kg^–1) were administered intramuscularly 15 minutes prior to laparoscopy. Heart rate (HR), respiratory rate, mean arterial pressure (MAP), peripheral oxygen saturation (SpO₂), end-tidal CO₂ partial pressure (Pe′CO₂), end-tidal concentration of isoflurane (Fe′Iso), pH, PaO₂, PaCO₂, bicarbonate (HCO₃^?), anion gap (AG) and base excess (BE) were monitored after chemical restraint, anesthesia induction and at different laparoscopy stages. Postoperative pain was assessed by visual analog scale (VAS) for 24 hours. Variables were compared using anova or Friedman test (p < 0.05).ResultsChemical restraint was effective in 92% of animals. Isoflurane anesthesia was effective; however, HR, MAP, pH and AG decreased, whereas Pe′CO₂, PaO₂, PaCO₂, HCO₃^? and BE increased. MAP was stable with tramadol and methadone treatments; HR, Fe′Iso and postoperative VAS decreased. VAS was lower for a longer time with methadone treatment; SpO₂ and AG decreased, whereas Pe′CO₂, PaCO₂ and HCO₃^? increased.Conclusions and clinical relevanceKetamine–midazolam provided satisfactory restraint. Isoflurane anesthesia for laparoscopy was effective but resulted in hypotension and respiratory acidosis. Tramadol and methadone reduced isoflurane requirements, provided postoperative analgesia and caused hypercapnia, with methadone causing severe respiratory depression. Thus, the anesthetic protocol is adequate for laparoscopy in Cuniculus paca; however, methadone should be avoided.     

Identification of Polymorphisms in the Osteopontin Gene and Their Associations with Certain Semen Production Traits of Water Buffaloes in the Brazilian Amazon

The osteopontin gene may influence the fertility of water buffaloes because it is a protein present in sperm. The aim of this work was to identify polymorphisms in this gene and associate them with fertility parameters of animals kept under extensive grazing. A total of 306 male buffaloes older than 18 months, from two farms, one in the state of Amapá and the other in the state of Pará, Brazil were used in the study. Seven SNPs were identified in the regions studied. The polymorphisms were in gene positions 1478, 1513 and 1611 in the region 5′upstrem and positions 6690, 6737, 6925 and 6952 in the region amplified in intron 5. The SNPs were associated with the traits, namely scrotal circumference, scrotal volume, sperm motility, sperm concentration and sperm pathology. There were significant SNPs (p < 0.05) for all the traits. The SNP 6690 was significant for scrotal circumference, sperm concentration, sperm motility and sperm pathology and the SNP 6737 for scrotal volume. The genotype AA of SNP 6690 presented the highest averages for scrotal circumference, sperm concentration and motility and the lowest total number of sperm pathologies. For the scrotal volume trait, the animals with the largest volume were correlated with the presence of the genotype GG of SNP 6737. These results indicate a significance of the osteopontin gene as it seems to exert a substantial influence on the semen production traits of male buffaloes.     

Factors affecting the prevalence and severity of footrot in a Merino flock selected for resistance to Haemonchus contortus

SUMMARY An outbreak of virulent footrot occurred in three lines of sheep selected for different levels of resistance to Haemonchus contortus , providing an opportunity to examine the factors affecting footrot severity. The selection lines did not differ significantly in average footrot score, but various other factors were found to have an effect. In breeding ewes, the average score was 10.2 (maximum possible score 20) and the prevalence of footrot was 96%. There was a significant age effect, with average score increasing from 9.1 ± 0.8 in 2-year-old ewes, to 13.4 ± 0.9 in 7-year-old ewes. Ewes that lost their lambs tended to have higher scores than those that either failed to lamb or successfully reared their litter. Significant differences in footrot score were caused by the lambing paddock in which the ewes had been, during the period seven weeks to one week before infection. Among lambs, aged about 20 weeks, the average footrot score was lower (5.0), as was the prevalence (75%). The score increased by an average of 0.09 ± 0.03 per day of age and the prevalence increased by an average of 0.6%± 0.3% per day of age. Lambs born and reared as twins had significantly lower scores than singles, and progeny of maiden dams had lower scores than progeny of older ewes. There was no significant difference because of the sex of the lamb. The paternal half-sib estimate of heritability of footrot score was 0.20 ± 0.14 and of the genetic correlation between footrot score and worm egg count was essentially zero (0.02 ± 0.20), but these estimates were probably biased by the confounding of sires with lambing paddocks. Nevertheless, the results strongly suggest that there are no adverse genetic associations between resistance to footrot and resistance to internal parasites.     

Epidemiological studies of pig diseases: 2. Post-weaning diarrhoea and performance in Western Australian pigs

Objective To determine in Australian pig herds the accuracy of French protocols for risk factor assessment of post-weaning diarrhoea and illthrift.
Procedure French protocols for the collection of data on health indicators and risk factors for post-weaning diarrhoea were conducted on 54 batches of weaner pigs from 28 Western Australian pig herds during three years.
Results Large variations in post-weaning performance were found. About one-third of the batches were growing at <200 g/day during the 3 weeks after weaning, and 54% had growth rates of <250 g/day. Weaning age and weight of at least 30 days and 7.9 kg, respectively, optimised weaner performance. Other risk factors associated with little post-weaning diarrhoea and good weaner performance were high creep feed intakes, relatively little diarrhoea as suckers, and, contrary to expectations, large temperature fluctuations.
Conclusion Overall, the 'predictability' of post-weaning problems as assessed by measurement of risk status, was good. However, the model was less accurate at predicting the performance of a single batch of pigs.     

The pathogenesis and treatment of osteomyelitis and laminitis in a stallion after the prolonged topical application of formalin to the distal phalanx

A 12-year-old Quarterhorse stallion was presented with a severe lameness in the left forefoot. There was a 3 cm diameter cavity in the sole that extended to the solar surface of the distal phalanx. Radiographs revealed an osteomyelitis and a sequestrum which probably developed following the prolonged topical application of 10 percent formalin. The sequestrum was removed and the infected bone curetted under general anaesthesia. The horse was shod with heart bar shoes on both front feet 7 days after the surgery. Eight months later, radiographs showed marked rotation of the distal phalanx despite continual shoeing with heart bar shoes. Surgical resection of the dorsal wall of the hoof at the toe removed the pressure applied by the laminar wedge, and combined with the stabilising and supporting action of the heart bar shoe, permitted realignment of the distal phalanx approximately 30 degrees closer to the normal skeletal axis by 26 days post-operatively. This case highlights some of the recent developments in the treatment of laminitis and suggests that effective treatment is possible if the value of the animal warrants the time and investment.