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Prevalence of Entamoeba gingivalis and Trichomonas tenax among patients with periodontal disease attending Dental Clinic, University College Hospital, Ibadan. 在伊巴丹大学学院医院牙科诊所就诊的牙周病患者中,齿龈内阿米巴和tenax毛滴虫的患病率。
Q3 Medicine Pub Date : 2023-07-01 Epub Date: 2023-09-18 DOI: 10.4103/tp.tp_8_23
Adenike Olatundun Oladokun, Paul Ogboru, Olanrewaju I Opeodu, Ahmed O Lawal, Mofolusho O Falade

Background: Periodontal disease is often caused by bacterial plaque. However, there are indications that some infective agents, including parasites, may play important roles in the pathogenesis of the disease.

Aim: This study aimed at assessing the prevalence of gingivitis and periodontitis, as well as the prevalence of Entamoeba gingivalis and Trichomonas tenax, in the oral biofilm of individuals with periodontal diseases using polymerase chain reaction.

Materials and methods: One hundred and six respondents with periodontal disease participated in the study. All study participants had a full-mouth examination, and dental plaque samples were collected with a sterile curette. Samples were transported to the laboratory in a cold chain and stored frozen till analyzed. DNA was extracted from the samples and amplified using EGO and ENTAM primers for Entamoeba and TGBK primers for T. tenax.

Results: The mean age of respondents was 45 ± 16.3 years, with none using tobacco. The prevalence of gingivitis and periodontitis obtained from this study was 84.9% and 15.1%. The prevalence obtained for Entamoeba and T. tenax was 0.9% each; however, no participant had both parasites. The positive samples were from male participants with moderate gingivitis.

Conclusion: Gingivitis was more prevalent than periodontitis, though with a high preponderance in females. E. gingivalis and T. tenax may not be of etiologic importance in periodontal disease as they occurred sparsely in the studied population.

背景:牙周病通常是由菌斑引起的。然而,有迹象表明,包括寄生虫在内的一些感染因素可能在疾病的发病机制中发挥重要作用。目的:本研究旨在使用聚合酶链式反应评估牙周病患者口腔生物膜中牙龈炎和牙周炎的患病率,以及牙龈内阿米巴和tenax毛滴虫的患病率。材料和方法:116名患有牙周病的受访者参与了这项研究。所有研究参与者都进行了全面的口腔检查,并用无菌刮匙收集牙菌斑样本。样品用冷链运输到实验室,冷冻保存直至分析。从样品中提取DNA,并使用用于内阿米巴的EGO和ENTAM引物以及用于T.tenax的TGBK引物进行扩增。结果:被调查者的平均年龄为45±16.3岁,没有人吸烟。从本研究中获得的牙龈炎和牙周炎的患病率分别为84.9%和15.1%。内阿米巴和T.tenax的患病率各为0.9%;然而,没有一名参与者同时感染了这两种寄生虫。阳性样本来自患有中度牙龈炎的男性参与者。结论:牙龈炎的发病率高于牙周炎,但女性发病率较高。牙龈E.givalis和T.tenax在牙周病中可能不具有病因重要性,因为它们在研究人群中很少发生。
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引用次数: 0
Multiple protozoal infections in a single immunocompromised patient: A case report. 一例免疫功能低下患者的多种原生动物感染:病例报告。
Q3 Medicine Pub Date : 2023-07-01 Epub Date: 2023-09-18 DOI: 10.4103/tp.tp_31_21
Rupak Chatterjee, Tanusree Mondal, Gautam Barik, Subhoroop Ghosh, Netai Pramanik, Ardhendu Maji, Tapashi Ghosh

Immunocompromised patients with human immunodeficiency virus (HIV) infection are prone to multiple infections, of which parasitic infections are an important cause. Parasitic protozoal infections - both by common and rare protozoa are documented in such patients. Here, we report a rare and interesting case of five protozoal infections affecting a single HIV-infected person at the same time of initial presentation. A 64-years-male came to us with complaints of chronic diarrhea for 6 months. He was investigated and found to be positive for HIV I. His stool examination revealed cysts of Entameba histolytica and Giardia lamblia and oocysts of Cryptosporidium species and Cystoisospora species. His toxoplasma IgG was also positive in high titer. The patient was medically diagnosed and was treated with medications as clinically prescribed - antiretroviral therapy was initiated and he was discharged in due course. A total of five protozoal infections were documented affecting a single person - newly diagnosed immunocompromised male, which by sheer qualitative count of patient case histories, indeed is a rare case reported in the medical literature.

人体免疫缺陷病毒(HIV)感染的免疫受损患者容易发生多种感染,其中寄生虫感染是一个重要原因。寄生性原生动物感染——常见和罕见的原生动物都记录在这类患者身上。在这里,我们报告了一个罕见而有趣的病例,五种原生动物感染在初次出现的同时影响一名HIV感染者。一位64岁的男性来找我们,抱怨慢性腹泻6个月。对他进行了调查,发现其HIV I呈阳性。他的粪便检查显示溶组织内阿米巴和兰氏贾第鞭毛虫囊肿,隐孢子虫和囊孢子虫卵囊。他的弓形虫IgG也呈高滴度阳性。该患者得到了医学诊断,并按照临床处方接受了药物治疗——开始了抗逆转录病毒疗法,并在适当时候出院。据记录,共有五种原生动物感染影响一个人——新诊断的免疫功能低下男性,从患者病史的定性统计来看,这确实是医学文献中报道的罕见病例。
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引用次数: 0
Neuroparasitology- A New Horizon. 神经病学——一个新的视野。
Q3 Medicine Pub Date : 2023-07-01 Epub Date: 2023-09-18 DOI: 10.4103/tp.tp_52_23
Subhash Chandra Parija
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引用次数: 0
Acute appendicitis secondary to Entamoeba histolytica infestation. 急性阑尾炎继发于溶组织内阿米巴感染。
Q3 Medicine Pub Date : 2023-07-01 Epub Date: 2023-09-18 DOI: 10.4103/tp.tp_47_21
Surbhi J Patel, Priya Pathak, Poornachandra Thejeswi, Manisha Bandari

Acute appendicitis secondary to amoebic infestation is a rare possibility, especially in endemic areas. If left untreated, significant postoperative morbidity and mortality can result from the spread of amoebic trophozoites and subsequent tissue reaction. Histopathological examination of the resected appendix with demonstration of invasion by the parasite is mandatory for establishing a diagnosis. Careful examination of the specimen is thus of paramount importance.

继发于阿米巴感染的急性阑尾炎是一种罕见的可能性,尤其是在流行地区。如果不及时治疗,阿米巴滋养层的扩散和随后的组织反应可能会导致显著的术后发病率和死亡率。必须对切除的阑尾进行组织病理学检查,并证明其受到寄生虫的侵袭,以确定诊断。因此,仔细检查标本至关重要。
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引用次数: 0
RNA interference-mediated hookworm control by gut-dwelling lactic acid bacteria. RNA干扰介导肠道乳酸菌控制钩虫。
Q3 Medicine Pub Date : 2023-07-01 Epub Date: 2023-09-18 DOI: 10.4103/tp.tp_74_22
Shaziya Bi, Manisha Sabnis, Anamika Singh, Sanjiban Kumar Banerjee

Background: The complex life cycle of geohelminth parasites make it difficult to manage repeated infections that occur in endemic areas, that requires the development of new therapeutic strategies. Human hookworm Ancylostoma sp. and Necator sp. are a serious problem in many areas of the world where the old age anti-helminthics are ineffective. To address this, we have tried a non-traditional therapeutic approach for a sustainable solution to manage parasite infections.

Aims and objectives: In this study, we attempted to develop a new type of therapy using value-added probiotics-producing antiparasitic RNA interference (RNAi) molecules against a vital hookworm (Ancylostoma sp.) enzyme, astacin-like metalloprotease Ac-MTP-1. A new gut delivery RNAi vector was designed to produce double-stranded RNA (dsRNA) against the target to be delivered by feeding with a probiotic Lactococcus lactis that when administered in endemic areas can potentially be used to control the spread of infection by interrupting the life cycle of hookworm.

Results: The engineered probiotics colonizing the gut, when consumed by the parasite released the dsRNA that which knocked down the target by RNAi interfering with their moulting and tissue migration. The initial penetration of the larvae into the gut lining was reduced by 70%, followed by a reduction of up to 50% in migration to the critical organs. The damage caused to the liver (30%-40%) and the kidneys (50%-60%) by the hookworm in mouse models as quantitated by enzymes released in the blood was totally reversed when the worms were pre fed with the engineered L. lactis before the parasite challenge.

Conclusions: We believe that this is the first demonstrated knockdown of a target gene in hookworms using probiotics genetically engineered to stimulate RNAi (RNAi food), administered through the oral route. This novel method of parasite control when extended to other hookworms, A. duodenalis and A. ceylanicum can augment the efficacy of the existing anthelminthics if combined with them.

背景:地蠕虫寄生虫的复杂生命周期使其难以控制在流行地区发生的反复感染,这需要开发新的治疗策略。人类钩虫钩虫属和Necator属是世界上许多地区的一个严重问题,在这些地区,老年抗蠕虫药物无效。为了解决这一问题,我们尝试了一种非传统的治疗方法,以找到一种可持续的解决方案来管理寄生虫感染。目的和目的:在这项研究中,我们试图开发一种新型的治疗方法,使用产生抗寄生虫RNA干扰(RNAi)分子的增值益生菌来对抗一种重要的钩虫(钩虫属)酶,类虾青素金属蛋白酶Ac-MTP-1。设计了一种新的肠道递送RNAi载体,通过喂食益生菌乳酸乳球菌来产生针对靶点的双链RNA(dsRNA),当在流行地区给药时,可以通过中断钩虫的生命周期来控制感染的传播。结果:当寄生虫食用时,定殖在肠道的工程益生菌释放出dsRNA,通过干扰其蜕皮和组织迁移的RNAi来敲低目标。幼虫对肠道内壁的初始渗透减少了70%,随后向关键器官的迁移减少了50%。通过血液中释放的酶定量,钩虫在小鼠模型中对肝脏(30%-40%)和肾脏(50%-60%)造成的损伤在寄生虫攻击前预先喂食工程乳杆菌时完全逆转。结论:我们认为,这是首次证明通过口服途径使用经基因工程改造的益生菌来刺激RNAi(RNAi食品)敲低钩虫的靶基因。当这种新的寄生虫控制方法推广到其他钩虫、十二指肠钩虫和马里兰钩虫时,如果与它们结合使用,可以提高现有驱虫剂的效力。
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引用次数: 0
Ascaris suum - A zoonosis in Bali, Indonesia. 蛔虫——印度尼西亚巴厘岛的一种人畜共患疾病。
Q3 Medicine Pub Date : 2023-07-01 Epub Date: 2023-09-18 DOI: 10.4103/tp.tp_24_23
Kadek Karang Agustina, I Made Ady Wirawan, I Made Sudarmaja, I Made Subrata, Nyoman Sadra Dharmawan

Background: Ascariasis is a common disease in humans and pigs. In previous reports, it has been known that ascariasis in humans is not only caused by Ascaris lumbricoides but also by Ascaris suum which is known to originate from pigs.

Aims and objectives: This research aims to identify the species of Ascaris that infect traditional pig farmers in Bali, Indonesia.

Materials and methods: To achieve this, an observational study was conducted, in which 239 feces samples were collected from traditional pig farmers. The collected samples were then divided into two parts, with one part stored in a pot containing 10% formalin for microscopic testing, and the other part stored in 70% ethanol for molecular testing. In cases where Ascaris infection was observed during the microscopic examination, a further test was conducted using the polymerase chain reaction (PCR) method for molecular examination. The DNA derived from the PCR was then subjected to DNA sequencing.

Results: The result was compared to A. suum gene data previously published in the Gene Bank. The results of the microscopic examination showed that 22 farmers were infected with Ascaris. However, PCR testing and DNA sequencing indicated that three infections were caused by A. suum.

Conclusion: Therefore, it can be concluded that A. suum is a zoonosis in Bali. Thus, efforts to control ascariasis must pay attention to the presence of pigs around it. Handling of ascariasis is not only carried out in humans but also in pigs and the environment.

背景:蛔虫是人和猪的常见疾病。在以前的报道中,已经知道人类的蛔虫不仅是由蛔虫引起的,而且是由猪引起的猪蛔虫引起的。目的和目的:本研究旨在确定感染印度尼西亚巴厘岛传统养猪户的蛔虫种类。材料和方法:为此,进行了一项观察性研究,从传统养猪户身上采集了239份粪便样本。然后将收集的样品分为两部分,一部分储存在含有10%福尔马林的罐中用于显微镜测试,另一部分储存于70%乙醇中用于分子测试。在显微镜检查中观察到蛔虫感染的情况下,使用聚合酶链式反应(PCR)方法进行进一步的分子检查。然后对源自PCR的DNA进行DNA测序。结果:该结果与之前发表在基因库中的A.suum基因数据进行了比较。显微镜检查结果显示,22名农民感染了蛔虫。然而,PCR检测和DNA测序表明,有三例感染是由苏艾引起的。结论:因此,可以得出结论,苏艾是巴厘岛的一种人畜共患病。因此,控制蛔虫的工作必须注意周围是否有猪。处理蛔虫不仅在人类身上进行,而且在猪和环境中也进行。
{"title":"<i>Ascaris suum</i> - A zoonosis in Bali, Indonesia.","authors":"Kadek Karang Agustina,&nbsp;I Made Ady Wirawan,&nbsp;I Made Sudarmaja,&nbsp;I Made Subrata,&nbsp;Nyoman Sadra Dharmawan","doi":"10.4103/tp.tp_24_23","DOIUrl":"10.4103/tp.tp_24_23","url":null,"abstract":"<p><strong>Background: </strong>Ascariasis is a common disease in humans and pigs. In previous reports, it has been known that ascariasis in humans is not only caused by <i>Ascaris lumbricoides</i> but also by <i>Ascaris suum</i> which is known to originate from pigs.</p><p><strong>Aims and objectives: </strong>This research aims to identify the species of <i>Ascaris</i> that infect traditional pig farmers in Bali, Indonesia.</p><p><strong>Materials and methods: </strong>To achieve this, an observational study was conducted, in which 239 feces samples were collected from traditional pig farmers. The collected samples were then divided into two parts, with one part stored in a pot containing 10% formalin for microscopic testing, and the other part stored in 70% ethanol for molecular testing. In cases where <i>Ascaris</i> infection was observed during the microscopic examination, a further test was conducted using the polymerase chain reaction (PCR) method for molecular examination. The DNA derived from the PCR was then subjected to DNA sequencing.</p><p><strong>Results: </strong>The result was compared to <i>A. suum</i> gene data previously published in the Gene Bank. The results of the microscopic examination showed that 22 farmers were infected with Ascaris. However, PCR testing and DNA sequencing indicated that three infections were caused by <i>A. suum</i>.</p><p><strong>Conclusion: </strong>Therefore, it can be concluded that <i>A. suum</i> is a zoonosis in Bali. Thus, efforts to control ascariasis must pay attention to the presence of pigs around it. Handling of ascariasis is not only carried out in humans but also in pigs and the environment.</p>","PeriodicalId":37825,"journal":{"name":"Tropical Parasitology","volume":"13 2","pages":"100-106"},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10583779/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49683191","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
An E-mail interview with Prof. Mohammed Hafeez. 穆罕默德·哈菲兹教授的电子邮件采访。
Q3 Medicine Pub Date : 2023-07-01 Epub Date: 2023-09-18 DOI: 10.4103/tp.tp_51_23
{"title":"An E-mail interview with Prof. Mohammed Hafeez.","authors":"","doi":"10.4103/tp.tp_51_23","DOIUrl":"10.4103/tp.tp_51_23","url":null,"abstract":"","PeriodicalId":37825,"journal":{"name":"Tropical Parasitology","volume":"13 2","pages":"129-132"},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10583786/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49683193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Human subcutaneous dirofilariasis: A diagnostic dilemma. 人类皮下丝虫病:诊断难题。
Q3 Medicine Pub Date : 2023-07-01 Epub Date: 2023-09-18 DOI: 10.4103/tp.TP_117_20
Parasmita Das Choudhury, Dina Raja, Vaishali Sarma

Human dirofilariasis is a rare infection caused by filarial worms of genus Dirofilaria. Humans may be infected as aberrant (accidental) hosts, mainly by Dirofilaria repens and Dirofilaria immitis. Most of the cases reported in India are of ocular dirofilariasis, with a few reports on subcutaneous dirofilariasis. We hereby report one such case of human subcutaneous dirofilariasis affecting the neck region of a 63-year male patient from Assam. This case indicates that this infection is a potentially emerging zoonosis in Assam and there is scope of finding more such cases with surveillance.

人丝虫病是一种罕见的由丝虫属丝虫引起的感染。人类可能作为异常(偶然)宿主被感染,主要是由白氏Dirofilaria repens和Imitis感染。印度报告的大多数病例是眼部dirofilasis,少数报告是皮下dirofilasia。我们在此报告一例来自阿萨姆邦的63岁男性患者的颈部受人皮下dirofilasis影响的病例。该病例表明,这种感染在阿萨姆邦是一种潜在的新出现的人畜共患疾病,通过监测可以发现更多此类病例。
{"title":"Human subcutaneous dirofilariasis: A diagnostic dilemma.","authors":"Parasmita Das Choudhury,&nbsp;Dina Raja,&nbsp;Vaishali Sarma","doi":"10.4103/tp.TP_117_20","DOIUrl":"10.4103/tp.TP_117_20","url":null,"abstract":"<p><p>Human dirofilariasis is a rare infection caused by filarial worms of genus <i>Dirofilaria</i>. Humans may be infected as aberrant (accidental) hosts, mainly by <i>Dirofilaria repens</i> and <i>Dirofilaria immitis</i>. Most of the cases reported in India are of ocular dirofilariasis, with a few reports on subcutaneous dirofilariasis. We hereby report one such case of human subcutaneous dirofilariasis affecting the neck region of a 63-year male patient from Assam. This case indicates that this infection is a potentially emerging zoonosis in Assam and there is scope of finding more such cases with surveillance.</p>","PeriodicalId":37825,"journal":{"name":"Tropical Parasitology","volume":"13 2","pages":"118-121"},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10583780/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49683202","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Primate malaria of human importance. 人类重要的灵长类疟疾。
Q3 Medicine Pub Date : 2023-07-01 Epub Date: 2023-09-18 DOI: 10.4103/tp.tp_79_22
Abhishek Mewara, Priya Sreenivasan, Sumeeta Khurana

Nonhuman primate (NHP) malaria poses a major threat to the malaria control programs. The last two decades have witnessed a paradigm shift in our understanding of the malaria caused by species other than the traditionally known human Plasmodium species - Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, and Plasmodium ovale. The emergence of the malaria parasite of long-tailed macaque monkeys, Plasmodium knowlesi, as the fifth malaria species of humans has made the scientific community consider the risk of other zoonotic malaria, such as Plasmodium cynomolgi, Plasmodium simium, Plasmodium inui, and others, to humans. The development of knowledge about P. knowlesi as a pathogen which was earlier only known to experimentally cause malaria in humans and rarely cause natural infection, toward its acknowledgment as a significant cause of human malaria and a threat of malaria control programs has been made possible by the use of advanced molecular techniques such as polymerase chain reaction and gene sequencing. This review explores the various aspects of NHP malaria, and the association of various factors with their emergence and potential to cause human malaria which are important to understand to be able to control these emerging infections.

非人灵长类(NHP)疟疾对疟疾控制计划构成了重大威胁。在过去的二十年里,我们对由传统已知的人类疟原虫以外的物种引起的疟疾的理解发生了范式转变——恶性疟原虫、间日疟原虫、疟疾疟原虫和卵圆疟原虫。长尾猕猴的疟原虫诺氏疟原虫作为人类第五种疟疾物种的出现,使科学界考虑了其他人畜共患疟疾对人类的风险,如食蟹痛疟原虫、猴疟原虫、菊粉疟原虫等。通过使用聚合酶链式反应和基因测序等先进的分子技术,诺氏疟原虫作为一种病原体的知识得以发展,这种病原体早期只在实验上引起人类疟疾,很少引起自然感染,并被公认为人类疟疾的重要原因和疟疾控制计划的威胁。这篇综述探讨了NHP疟疾的各个方面,以及各种因素与它们的出现和导致人类疟疾的潜力的关系,这对于理解能够控制这些新出现的感染很重要。
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引用次数: 0
Parafilm as an efficient transport matrix for corneal scrapings. Parafilm作为角膜刮伤的有效传输基质。
Q3 Medicine Pub Date : 2023-07-01 Epub Date: 2023-09-18 DOI: 10.4103/tp.tp_67_22
Chayan Sharma, Sumeeta Khurana
Introduction: Acanthamoeba spp. are free-living parasites increasingly implicated in causing Acanthamoeba keratitis (AK). AK is diagnosed by demonstration of parasites in corneal samples by direct microscopy, culture, and nucleic acid amplification. Most commonly, corneal scrapings are sent to the laboratory smeared between two glass slides. These scrapings are suitable for direct microscopy but less suitable for culture and polymerase chain reaction (PCR) which, in turn, are more sensitive for the diagnosis of AK. Aim: The aim of the study was to explore better alternatives for transporting corneal scrapings from the point-of-care eye center to the concerned laboratories. Materials and Methods: The study used small Parafilm (Bemis Company Inc., USA) squares (PSs) of 1 cm each prepared by cutting Parafilm using a surgical blade under sterile conditions. Each of the four different dilutions of Acanthamoeba suspension (15, 30, 60, and 120 cells) was used in this study. Each dilution was added onto the surface of 36 PSs and kept at room temperature for 24-h, 48-h, and 72-h incubation. The PSs for one particular time point and dilution were used for calcofluor white staining, its inoculation onto the surface of nonnutrient agar having a lawn of Escherichia coli, and Acanthamoeba-specific PCR amplification. In addition, two PSs inoculated with 30 cells and incubated for 24 h and 72 h were used for scanning electron microscopy (SEM). Results and Conclusion: All three diagnostic techniques, i.e. microscopy, culture, and PCR, detected the presence of Acanthamoeba at all the tested concentrations and time points. However, the growth pattern on culture changed directly in proportion to increased incubation periods and increased concentration of inoculum. In addition, the adherence of Acanthamoeba to the Parafilm was confirmed by SEM; these results suggest the use of these PSs as a suitable matrix for the transport of corneal scrapings.
简介:棘阿米巴是一种自由生活的寄生虫,越来越多地与引起棘阿米巴角膜炎(AK)有关。AK是通过直接显微镜、培养和核酸扩增在角膜样本中显示寄生虫来诊断的。最常见的是,角膜刮片被送到实验室,涂抹在两张载玻片之间。这些刮片适用于直接显微镜检查,但不太适用于培养和聚合酶链式反应(PCR),后者反过来对AK的诊断更敏感。目的:本研究的目的是探索将角膜刮片从护理点眼科中心运送到相关实验室的更好替代方案。材料和方法:该研究使用1厘米的小Parafilm(Bemis Company股份有限公司,USA)正方形(PS),每个正方形通过在无菌条件下使用手术刀片切割Parafilm而制备。本研究中使用了四种不同稀释度的棘阿米巴悬浮液(15、30、60和120个细胞)。将每个稀释液添加到36个PS的表面上,并在室温下保持24小时、48小时和72小时孵育。使用一个特定时间点的PS和稀释液进行钙荧光白染色,将其接种到具有大肠杆菌草坪的非营养琼脂表面,并进行棘阿米巴特异性PCR扩增。此外,用30个细胞接种并孵育24小时和72小时的两个PS进行扫描电子显微镜(SEM)。结果和结论:显微镜、培养和PCR三种诊断技术在所有测试浓度和时间点都检测到棘阿米巴的存在。然而,培养物上的生长模式与培养时间的增加和接种物浓度的增加成正比。此外,扫描电镜证实了棘阿米巴对Parafilm的粘附性;这些结果表明使用这些PS作为运输角膜刮片的合适基质。
{"title":"Parafilm as an efficient transport matrix for corneal scrapings.","authors":"Chayan Sharma,&nbsp;Sumeeta Khurana","doi":"10.4103/tp.tp_67_22","DOIUrl":"10.4103/tp.tp_67_22","url":null,"abstract":"Introduction: Acanthamoeba spp. are free-living parasites increasingly implicated in causing Acanthamoeba keratitis (AK). AK is diagnosed by demonstration of parasites in corneal samples by direct microscopy, culture, and nucleic acid amplification. Most commonly, corneal scrapings are sent to the laboratory smeared between two glass slides. These scrapings are suitable for direct microscopy but less suitable for culture and polymerase chain reaction (PCR) which, in turn, are more sensitive for the diagnosis of AK. Aim: The aim of the study was to explore better alternatives for transporting corneal scrapings from the point-of-care eye center to the concerned laboratories. Materials and Methods: The study used small Parafilm (Bemis Company Inc., USA) squares (PSs) of 1 cm each prepared by cutting Parafilm using a surgical blade under sterile conditions. Each of the four different dilutions of Acanthamoeba suspension (15, 30, 60, and 120 cells) was used in this study. Each dilution was added onto the surface of 36 PSs and kept at room temperature for 24-h, 48-h, and 72-h incubation. The PSs for one particular time point and dilution were used for calcofluor white staining, its inoculation onto the surface of nonnutrient agar having a lawn of Escherichia coli, and Acanthamoeba-specific PCR amplification. In addition, two PSs inoculated with 30 cells and incubated for 24 h and 72 h were used for scanning electron microscopy (SEM). Results and Conclusion: All three diagnostic techniques, i.e. microscopy, culture, and PCR, detected the presence of Acanthamoeba at all the tested concentrations and time points. However, the growth pattern on culture changed directly in proportion to increased incubation periods and increased concentration of inoculum. In addition, the adherence of Acanthamoeba to the Parafilm was confirmed by SEM; these results suggest the use of these PSs as a suitable matrix for the transport of corneal scrapings.","PeriodicalId":37825,"journal":{"name":"Tropical Parasitology","volume":"13 2","pages":"84-88"},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10583775/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49683206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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Tropical Parasitology
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