利什曼原虫与黑热病

利什曼原虫 (Leishmania)是一类微小的鞭毛虫 ,寄生于人体的有 3种——杜氏利什曼原虫、热带利什曼原虫和巴西利什曼原虫。在其发现之前 ,临床上发现其致病已有数百年历史。在印加前期陶制人佣上所绘的鼻咽病灶 ,据鉴定就是美洲利什曼病。 1898年 ,一俄国军医已作过精确描述。这 3种原虫以杜氏利什曼原虫危害最大。杜氏利什曼原虫寄生于人体的肝、脾、骨髓和淋巴腺等细胞之内 ,虫体圆形或卵圆形 ,只有 4 .2μm× 2 .8μm大小 ,它能引起黑热病 ,又名黑热病原虫。其生活史有 2个阶段 :一个阶段寄生于人体 (或狗 ) ,另一个阶段寄生在白蛉子体…     

我国人与犬、猫共患病防控存在的主要问题与对策建议

随着社会经济的发展、城市化进程的加速及人们生活水平的不断提高,越来越多的宠物走进了人们的家庭,在人类社会生活中发挥了积极作用。但同时,宠物也是人兽共患病的重要传染源和传播媒介。与人类关系最为密切的犬、猫在人兽共患病的防控中具有重要的意义。在已报道的200多种主要的人兽共患病中,与宠物犬、猫有直接或间接关系的有70余种。随着宠物犬猫数量的大幅攀升和宠物业的飞速发展,我国人与犬猫共患病可能会出现逐步高发的趋势,疫病防控工作面临着许多问题。为了完善宠物管理制度,建立有效的防疫监督体系,对人与犬猫共患病实行有效的防控,本文就完善法律法规、形成综合管理机制,加强卫生监督、强化无害化处理、培养专业人才以及广泛开展宣传教育等6个方面提出了相关建议。     

新型冠状病毒体外诊断技术研发现状与展望

随着新型冠状病毒(SARS-CoV-2)引发的全球疫情不断加剧,新型冠状病毒体外诊断技术与产品的研发成为全球生物医药领域关注的重点,体外诊断技术与产品的研发创新对于提升新发突发传染病的防控能力有着重要的意义.对2019年12月至2020年12月期间国内外新型冠状病毒体外诊断技术与产品的研发现状与发展趋势进行了综述,即包...     

野猪与人冲突防控对策研究的系统评价

在中国,野猪(Sus scrofa)与人冲突问题亟需采取有效、科学的对策进行防控。通过了解野猪与人冲突防控对策的文献数量、研究范围、防控对策的种类及有效性等方面,对国内外野猪与人冲突防控对策进行系统分析与评价,以期为我国防控野猪与人冲突提供理论依据。检索中英文数据库建库至2021年5月31日的文献,提取文献的研究起始年份、研究所在地理单元、使用的防控措施种类、措施是否有效、措施的具体内容评价等。共搜集文献446篇,最终纳入77篇,分别来自欧洲、亚洲、北美洲、大洋洲、南美洲五大洲,研究时间主要在1981年后。防控对策类型主要分为致死性防控(药物、猎捕)及非致死性防控(干扰技术、设置障碍物、补充喂养、生育控制、生物防控)两大类;致死性防控对策的有效性(81.5%)高于非致死性防控对策的有效性(68.4%)。相比于非致死性防控对策,致死性防控对策是目前有效性较高的防控策略,而猎捕是现阶段调控野猪种群可行性较高的主要致死性防控对策。国外在猎捕方面的经验可为我国开展野猪防控工作提供参考。     

吉林珲春地区野猪危害防控研究

近年来人与野生动物冲突日趋严重。2012年7月到2014年10月,利用东北虎(Panthera tigris altaica)粪便、声音在吉林珲春地区开展了野猪(Sus scrofa)危害防控研究。以采取防控措施起始时间到野猪进入实验样地或对照样地的时间间隔作为防控有效期;以实验结束时实验样地内放置东北虎粪便、东北虎声音播放器位置或对照样地中心点与样地内野猪危害发生处的最短距离作为防控范围。将防控有效期和防控范围分别转化为有效期指数和范围指数。结果表明:(1)东北虎粪便对野猪危害防控效果显著,与对照组相比,有效期指数和范围指数均存在显著差异(P0.05);(2)东北虎声音对野猪危害防控效果显著,与对照组相比,有效期指数和范围指数均差异显著(P0.05);(3)同时应用东北虎粪便和声音防控野猪危害效果显著,与对照组相比,有效期指数和范围指数均差异显著(P0.05);(4)单独应用东北虎粪便、单独应用东北虎声音与同时应用东北虎粪便和声音的各种防控方法之间,有效期指数和范围指数无显著差异(P0.05)。本研究认为,东北虎粪便、声音对野猪危害防控效果显著与东北虎是野猪天敌,且珲春地区存在东北虎有关。     

当前传染病防控工作面临形势与分析

传染病发病突然、传播较广和迅速,对社会、政治、经济等方面影响巨大,是严重危害人民身体健康和生命安全的疾病,是全人类第二位的致死原因。传染病防控工作关系到人民健康和社会和谐安定,预防及控制传染病的发生和流行的任务十分艰巨,是新时期乃至今后相当长一段时间疾控工作的重中之重。     

蝗虫成灾规律、影响因素及防控技术研究进展

蝗虫是最重要的全球农牧业害虫之一,其灾害不仅强烈影响生态系统功能和稳定性,还带来严重的经济损失。因蝗虫食性杂、适应性强、聚集为害等特点使其可持续治理成为一项复杂而长期的艰巨工作。本文从蝗虫灾变发生规律、影响因素(自身因素、非生物和生物因素)以及当前有效防控方法与技术进行综述,并从蝗虫种群动态变化规律以及其有效防控的理论与技术提出今后的研究方向,为将来蝗虫发生的内在机制研究以及对其进行绿色防控技术探索提供基础。     

宠物狂犬病及其防控

本文对狂犬病在犬、猫等宠物中的流行情况;宠物狂犬病及其危害;宠物狂犬病的预防及国外宠物狂犬病的防控经验等进行了综述,为保证宠物和人类健康、有效防控宠物狂犬病提供参考。     

Heme metabolism in promastigotes of Leishmania donovani

Promastigotes of Leishmania donovani (Dd-8 strain) showed presence of important key enzymes of heme synthesizing (d-aminolevulinic acid synthase and ferrochelatase) and degrading (heme oxygenase and biliverdin reductase) systems, classical leishmanicidal drugs viz allopurinol, amphotericin B, pentamidine and CDRI compound 93/202 inhibited the heme oxygenase activity of the parasite, whereas, -aminolevulinic acid synthase activity practically remained unaffected. The Km, Vmax ad pH values of heme oxygenase of promastigotes were found to be 1666 M hemin, 625 nmol of bilirubin formed h-1 mg protein-1 and 7.5 respectively. The findings suggest the presence and importance of heme metabolism in the de novo synthesis of different hemoproteins of the Leishmania parasite as well as the detoxification and its defence against biological insults.     

Phosphofructokinase of Leishmania donovani and Leishmania braziliensis and its Role in Glycolysis*

SYNOPSIS. It was shown in an investigation of the phosphofructokinases of Leishmania donovani and Leishmania braziliensis that both enzymes are similar to that of Crithidia fasciculata. Although the enzymes are allosteric with respect to their substrates and require AMP for activation, there is no influence by other heterotropic modifiers. The Mg²⁺-ATP chelate activates these enzymes in a first order process and they can be inhibited by free ATP. The inhibition is reversed by the activator, AMP, in a competitive manner. The requirement for the nucleotide in L. donovani can be eliminated by decreasing the pH. The data indicate that phosphofructokinase, a pivotal enzyme in glycolysis for most organisms, probably does not play an important role in glycolysis in Leishmania.     

Effects of Cyclophosphamide on Visceral Leishmaniasis in the Mouse*

SYNOPSIS Cyclophosphamide (Cy), 125 or 200 mg/kg body weight, injected intraperitoneally (i.p.) into BALB/c mice one day before infection with amastigotes of Leishmania donovani, by the 8th day postinfection caused a significant decrease in the mean numbers of parasites in spleens and livers when compared to mice injected with phosphate buffered saline (PBS). When 125 mg/kg was injected into chronically infected mice (on day 34 of infection), however, within 2 days (day 36) mean parasite levels in both the spleens and livers were statistically greater than in PBS-treated controls. Similarly, when a series of 6 Cy injections, 50 mg/kg each, was injected over a period of 8 days during the chronic stage of infection, the mean parasite levels in both spleens and livers were significantly increased over those of PBS-treated controls. Druing the chronic stage of infection, Cy injections suppressed the immunity to superinfection. Neither plasma nor parasitized peritoneal macrophages obtained from Cy-treated mice, when compared to PBS-treated mice, differed in their respective capacities to influence the growth of intracellular amastigote of L. donovani in vitro. Passive transfer of hyperimmune mouse serum could not reverse the immunosuppressive effects of Cy upon chronic leishmaniasis in the mouse. It is suggested that neither readily demonstrable anti-leishmanial humoral factors nor “immune” macrophages per se, plays a major role in acquired immunity to leishmaniasis in the mouse.     

Effect of the microtubule stabilising agent taxol on leishmanial protozoan parasites in vitro

Taxol, a mitotic spindle toxin, was found to selectively inhibit the proliferation of Leishmania donovani in vitro at nanomolar concentrations with an IC50 of 35 nM. Concentrations of taxol as high as 50 nM, however, did not affect J774A.1 murine macrophages. Taxol (30 nM) also inhibited amastigote multiplication within a J774A.1 macrophage cell line when used in a 10-day experiment. It resulted in the in vitro assembly of L. donovani microtubules in a dose-dependent manner. When promastigotes were exposed to different concentrations of taxol for 24 h, cells were largely blocked in the G2-M phase of the cell cycle and there was a marked reduction in the percentage of cells in the S phase. The selective nature of taxol action against the parasite and its effectiveness in controlling amastigote multiplication emphasise its use as a promising chemotherapeutic against kala-azar.     

Trypsin and chymotrypsin-Iike enzymes of the sandfly Phlebotomus papatasi infected with Leishmania and their possible role in vector competence

Phlebotomus papatasi (Scopoli) is susceptible to infection with Leishmania major Yakimov & Schokov and resistant to L. donovani Laveran & Mesnil. The possibility that susceptibility depends on midgut levels of trypsin and chymotrypsin-like (esterolytic) enzymes was investigated. Infection with L. major reduced the trypsin-like activity to 93.5% and 86% of the control value at 20 and 30 h post feeding and increased it to 106% at 52 h. Infection with L. donovani reduced trypsin-like activity to 64% and 73% of the control value at 30 and 52 h post feeding. The overall amount of trypsin and chymotrypsin-like enzymes in L. major infections was reduced to 50% and 34% of the control value at 20 and 30 h post feeding and increased to 184% at 52 h. Only one of the enzymes separated by gel electrophoresis was lower throughout, i.e. peak D. Overall, the midgut enzyme level with L. donovani infection was 86% of the control value at 30 h post feeding and 105% at 52 h; their relative amounts changed throughout. Soybean trypsin inhibitor enabled L. donovani to survive and multiply in P. papatasi. It is suggested that a specific component of the trypsin-like activity prevents the survival of L. donovani in P. papatasi and that modulation of this factor enables L. major to survive.     

Acid phosphatase activity of promastigotes of Leishmania donovani: a marker of virulence

Seven cloned lines of promastigotes of Leishmania donovani (UR 6) were isolated by limiting dilution. One clone, UR6-C25, failed to multiply inside the macrophages of line J774G8 and thus was labelled as avirulent. Another, UR6-C24, multiplied inside macrophages, had a virulence index as high as 93 +/- 9.8 and was thus labelled as highly virulent. The other five clones had variable degree of virulence indices ranging from 46.4 +/- 5.8 to 67.6 +/- 3.5. No significant difference in the degrees of attachment of virulent and avirulent populations of promastigotes to macrophages was observed, suggesting no difference in the ligand utilised by these populations for attachment to the macrophages. Acid phosphatase activity of cloned promastigotes correlated with the degree of virulence. These data suggest that acid phosphatase activity could be used as a marker to differentiate avirulent from virulent populations of promastigotes of L. donovani.     

Cultivation of Leishmania donovani and Leishmania braziliensis in Defined Media: Nutritional Requirements

SYNOPSIS Nutritional requirements of promastigotes of Leishmania donovani and Leishmania braziliensis were studied in modifications of a simple defined culture medium. "Continuous growth," considered as propagation through 10 successive passages, was supported by inorganic salts, 14 l -amino acids (arginine, cysteine, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, threonine, tryptophan, tyrosine, valine), glucose, adenosine, and a mixture of 11 vitamins and related growth factors. Purified defatted bovine serum albumin proved beneficial. The nutritional needs of the above species of Leishmania differ from those of 2 other hemoflagellate species, Leishmania tarentolae and Crithidia fasciculata , for which glucose, proline and glutamine were found to be nonessential. It is suggested that lower hemoflagellates may be capable of synthesizing these substrates de novo. Leishmania donovani and L. braziliensis required higher levels of folic acid than L. tarentolae , probably due to the fact that folates are involved as cofactors in the biosyntheses of pyrimidines and serine. Although the mixtures reported here cannot be regarded as "minimal essential" media, they are considerably less complex than the ones employed so far for cultivating hemoflagellates, and are therefore well suited for studies related to nutrition and biosynthetic capabilities of Trypanosomatids.     

Identification of Leishmania spp. by Radiorespirometry

SYNOPSIS Preliminary investigation of the application of radiorespirometric technic to protozoan parasites of man indicates a potential for rapid identification. This technic, developed for identification of bacteria, was modified for use with culture forms of Leishmania. Five strains of Leishmania were compared: 2 of L. donovani , 2S and K; L. brasiliensis , 2936 and B; and 1 of L. tropica , A. Consistent and rapid (2 hr) identification was obtained by the radiorespirometric procedure. A computer-type analysis of the radiorespirometric profiles of the 5 strains permitted correct identification of each isolate at the strain level 100% of the time. This technic offers several advantages over many current procedures for identification of protozoan parasites: (A) It is simple, rapid and highly reproducible. (B) Since it does not rely on visual or spectrophoto-metric determination, it may be conducted in the presence of optically complex substances. (C) It requires relatively low numbers of organisms (2 × 10⁵/¹⁴C-labeled substrate). (D) It is based on differential enzymic activity between species and strains of organisms and therefore, ultimately, on inherent genetic determinates of the parasites. (E) Further development of the procedure and accumulation of a data reference "bank" would allow automation of most of the identification process.     

Isolation of a taxol-resistant Leishmania donovani promastigote mutant that exhibits a multidrug-resistant phenotype

We raised a strain of Leishmania donovani in the laboratory that was resistant to 500 nM taxol. The IC50 of the wild-type strain for taxol was 35 nM and that of the taxol-resistant strain (T-500) was 1 microM. The T-500 strain exhibited a Mdr phenotype; it was also resistant to other unrelated drugs like vinblastine, adriamycin and the commonly used antimonial drugs pentostam and glucantime. Verapamil (20 nM), a calcium channel blocker, was found to reverse the resistance of T-500 to taxol. Acquired resistance to taxol has been reported to be mediated by alterations involving tubulin in cancer cells. Thus polymerisation assays with tubulin fractions in wild-type versus taxol-resistant cells (T-500) were performed in vitro. The tubulin fraction from T-500 was more resistant to in vitro polymerisation than the tubulin isolated from the wild-type, suggesting that this is one means by which the parasite may acquire resistance to taxol.     

Polymerase chain reaction detection of Leishmania DNA in skin biopsy samples in Sri Lanka where the causative agent of cutaneous leishmaniasis is Leishmania donovani

Leishmania donovani is the known causative agent of both cutaneous (CL) and visceral leishmaniasis in Sri Lanka. CL is considered to be under-reported partly due to relatively poor sensitivity and specificity of microscopic diagnosis. We compared robustness of three previously described polymerase chain reaction (PCR) based methods to detectLeishmania DNA in 38 punch biopsy samples from patients presented with suspected lesions in 2010. Both, Leishmaniagenus-specific JW11/JW12 KDNA and LITSR/L5.8S internal transcribed spacer (ITS)1 PCR assays detected 92% (35/38) of the samples whereas a KDNA assay specific forL. donovani (LdF/LdR) detected only 71% (27/38) of samples. All positive samples showed a L. donovani banding pattern upon HaeIII ITS1 PCR-restriction fragment length polymorphism analysis. PCR assay specificity was evaluated in samples containing Mycobacterium tuberculosis, Mycobacterium leprae, and human DNA, and there was no cross-amplification in JW11/JW12 and LITSR/L5.8S PCR assays. The LdF/LdR PCR assay did not amplify M. leprae or human DNA although 500 bp and 700 bp bands were observed in M. tuberculosis samples. In conclusion, it was successfully shown in this study that it is possible to diagnose Sri Lankan CL with high accuracy, to genus and species identification, using Leishmania DNA PCR assays.