cats

  • 文章类型: Journal Article
    猫泛白细胞减少症是由猫泛白细胞减少症病毒(FPV)引起的一种传染性病毒性疾病。一种密切相关的病原体是犬细小病毒(CPV),和该病毒中的氨基酸取代使其能够获得猫科动物的宿主范围。在猫科动物宿主中,由CPV引起的疾病表现出与FPV或较温和的症状相似的症状,导致其诊断不足。这项研究的目的是确定具有泛白细胞减少症临床症状的猫中CPV2型(CPV-2)的存在,并评估将商业CPV抗原测试用于FPV的临床诊断。
    来自斯洛伐克中部的59只猫的样品被包括在研究中。收集直肠拭子并使用商业抗原测试对细小病毒感染进行临床测试。通过靶向病毒VP2基因的PCR确认抗原阳性样品。用Sanger法建立PCR产物的序列。
    在59个样本中,通过抗原和PCR测试发现23例细小病毒感染呈阳性(38.9%)。国家生物技术信息中心BLASTn应用的分析显示与FPV成对同一性99.78-100%。本研究中纳入的细小病毒感染猫的死亡率为8.69%(2/23)。
    尽管未证实患有CPV-2的猫科动物疾病,CPV抗原检测能够检测FPV感染.
    UNASSIGNED: Feline panleukopenia is a contagious viral disease caused by the feline panleukopenia virus (FPV). A closely related pathogen is canine parvovirus (CPV), and amino acid substitutions in this virus allow it to acquire a feline host range. In feline hosts, the disease induced by CPV manifests with similar symptoms to those caused by FPV or milder ones, leading to its underdiagnosis. The aim of this study was to determine the presence of CPV type 2 (CPV-2) in cats with clinical symptoms of panleukopenia and to assess the use of commercial CPV antigen tests for the clinical diagnosis of FPV.
    UNASSIGNED: Samples from 59 cats from central Slovakia were included in the study. Rectal swabs were collected and clinically tested for parvovirus infection using a commercial antigen test. Antigen-positive samples were confirmed by PCR targeting the viral VP2 gene. The sequences of the PCR products were established with the Sanger method.
    UNASSIGNED: Of 59 samples, 23 were revealed to be positive for parvovirus infection by both antigen and PCR test (38.9%). Analysis with the National Center for Biotechnology Information BLASTn application showed 99.78-100% pairwise identity with FPV. The mortality rate of parvovirus-infected cats included in this study was 8.69% (2/23).
    UNASSIGNED: Although feline disease with CPV-2 was not confirmed, the CPV antigen test was able to detect FPV infection.
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  • 文章类型: Journal Article
    淋巴瘤是家猫最常见的肿瘤,最常发生在小肠。猫小肠淋巴瘤主要显示通过具有CD3的T细胞的标准免疫阳性或具有CD20的B细胞的标准免疫阳性鉴定的T细胞免疫表型。相比之下,根据WHO分类,在人类中应用了广泛的免疫组织化学抗体来诊断各种特定的淋巴瘤亚型。我们的目的是增加我们对在肠道中形成宏观肿块的猫科动物非B细胞淋巴瘤的免疫表型的了解。我们评估了15例患者的免疫组织化学和流式细胞术联合发现。93%(14/15)的肿瘤淋巴样细胞对CD3免疫阳性,颗粒酶B占87%(13/15),CD5在20%(3/15),13%的CD8(2/15),7%(1/15)的CD4和7%(1/15)的CD56病例。仅在13%(2/15)的病例中通过组织病理学和在47%(7/15)的病例中通过细胞学鉴定了表明肿瘤细胞的细胞毒性起源的细胞毒性颗粒。没有细胞毒性蛋白颗粒酶B的免疫组织化学标记,在46%(6/13)的细胞学切片和85%(11/13)的组织病理学切片中,细胞毒性状态会被遗漏.这些发现表明,更复杂的免疫表型可能会增进我们的理解,并有助于预测猫的小肠T细胞淋巴瘤。
    Lymphoma is the most common tumour of domestic cats, developing most frequently in the small intestine. Feline small intestinal lymphoma predominantly demonstrates a T-cell immunophenotype identified by standard immunopositivity for T cells with CD3 or immunopositivity for B cells with CD20. In contrast, a wide spectrum of immunohistochemical antibodies are applied in humans to diagnose the various specific lymphoma subtypes according to the WHO classification. Our aim was to augment our knowledge of immunophenotypes in feline non-B-cell lymphomas forming macroscopic masses in the intestinal tract. We evaluated the combined immunohistochemistry and flow cytometry findings from 15 cases. Neoplastic lymphoid cells were immunopositive for CD3 in 93% (14/15), granzyme B in 87% (13/15), CD5 in 20% (3/15), CD8 in 13% (2/15), CD4 in 7% (1/15) and CD56 in 7% (1/15) of cases. Cytotoxic granules indicating a cytotoxic origin of the neoplastic cells were identified by histopathology only in 13% (2/15) and by cytology in 47% (7/15) of the cases. Without immunohistochemical labelling of the cytotoxic protein granzyme B, the cytotoxic status would have been missed in 46% (6/13) of the cytological and in 85% (11/13) of the histopathological slides. These findings suggest that more complex immunophenotyping may advance our understanding and help prognosticate small intestinal T-cell lymphoma in cats.
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  • 文章类型: Journal Article
    Opisthorchisviverrini感染是东南亚农村地区的一个紧迫的健康问题,并且与胆管癌的风险有关。尽管控制努力,高感染率持续存在,包括治疗后再感染的证据。这项研究旨在通过在3年的时间内在泰国农村流行地区采取综合的“单一健康”方法来解决这一公共卫生问题。该研究包括来自Udon他尼省的3600名参与者的数据,泰国,在2020年至2022年期间,涉及综合流行病学数据收集和危险因素分析,以了解各种干预措施对社区疾病传播的影响。通过监测2021年和2022年O.Viverrini再感染的发生率来评估干预措施的有效性。2020年,确认了218例O.viverrini感染(6.0%)。重要的风险因素包括靠近水体和食用生鱼。影响参与者感染风险的变量(P<0.001)是教育水平,参与传统仪式,卫生条件差,附近的水体中没有鸭子,寄生虫的自我药物治疗,和家庭中的多种感染。狗,猫,鲤鱼的患病率为5.4%,6.3%,11.5%,分别。地理分析显示,水体周围有成群的受感染家庭。干预措施,包括深度访谈,焦点小组讨论,健康教育,驱虫治疗,和使用当地自由放养鸭子的生物防治,实施了,第二年没有人再感染,第三年的患病率最低为0.3%。这项研究为感染流行率的动态变化提供了有价值的见解,为有效的疾病控制和社区健康促进做出重大贡献。这种综合的“一个健康”方法被证明是预防和控制opisthorchiasis的有效策略。
    Opisthorchis viverrini infection is a pressing health issue in rural Southeast Asia and is associated with the risk of cholangiocarcinoma. Despite control efforts, high infection rates persist, including evidence of reinfection post-treatment. This study aimed to address this public health concern through an integrated One-Health approach in endemic areas in rural Thailand over a 3-year period. The study included data from 3600 participants from Udon Thani Province, Thailand, during the years 2020 to 2022 and involved integrated epidemiological data collection and risk factor analysis to understand the impact of various interventions on disease transmission in the community. The efficacy of interventions was assessed by monitoring the incidence of O. viverrini reinfection in 2021 and 2022. In 2020, 218 cases of O. viverrini infection (6.0%) were identified. Significant risk factors included proximity to water bodies and consumption of raw fish. Variables contributing to infection risk among participants (P < 0.001) were education level, engagement in traditional ceremonies, poor sanitation, absence of ducks in nearby water bodies, self-medication for parasitic conditions, and multiple infections within a household. Dogs, cats, and cyprinoid fish showed prevalence rates of 5.4%, 6.3%, and 11.5%, respectively. Geographic analysis revealed clusters of infected households around water bodies. Interventions, including in-depth interviews, focus-group discussions, health education, anthelminthic treatment, and biological control using local free-range ducks, were implemented, resulting in no human reinfections in the second year and a minimal 0.3% prevalence rate in the third year. This study offers valuable insights into the dynamic changes in infection prevalence, making a significant contribution to effective disease control and community health promotion. This integrated One-Health approach proved to be an effective strategy for the prevention and control of opisthorchiasis.
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  • 文章类型: Journal Article
    形态异常被认为是自然蜱种群中的罕见现象。正在描述新的蜱异常病例,比如身体伪君子,纳米主义,性腺畸形和肢体畸形。从猫身上取出的蜱在形态上被鉴定为物种和发育阶段。确定在宿主上喂食的时间。使用PCR和Real-TimePCR方法对标本进行了测试,以确定是否存在常见的tick传播病原体:吞噬细胞无性体,巴贝西亚,疏螺旋体属。,米库伦希特氏菌,立克次体属。为了可视化异常结构,进行扫描电子显微镜(SEM)。the虫被鉴定为I.ricinus的成年雌性,在idiosoma的左侧表现出i腿的外行。该标本对两种医学上重要的病原体进行了阳性检测:吞噬细胞菌根和N.mikurensis。病例报告描述了来自波兰的I.ricinustick的形态异常的罕见病例。
    Morphological anomalies are considered a rare phenomenon among natural tick populations. New cases of abnormalities in ticks are being described, such as body assymetries, nanism, gynandromorphism and limb malformations. The tick removed from a cat was morphologically identified to species and developmental stage. The time of feeding on the host was determined. The specimen was tested using PCR and Real-Time PCR methods for the presence of the common tick-borne pathogens: Anaplasma phagocytophilum, Babesia spp, Borrelia spp., Neoehrlichia mikurensis, Rickettsia spp. For visualisation of the anomalous structures, scanning electron microscopy (SEM) was performed. The tick was identified as a slightly engorged adult female of I. ricinus exhibiting ectromely of leg I on the left side of the idiosoma. The specimen was tested positive for two medically important pathogens: A. phagocytophilum and N. mikurensis. The case report describes a rare case of a morphological anomaly in an I. ricinus tick from Poland.
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  • 文章类型: Journal Article
    目的:大肠埃希菌是狗和猫尿路感染(UTI)尿样中最常见的细菌之一。狗和猫的简单UTI可以用短期一线抗菌药物治疗,例如阿莫西林,阿莫西林与克拉维酸,或者甲氧苄啶/磺胺。复发性或复杂性UTI通常需要使用广谱抗生素进行长期治疗。然而,药物的选择应基于抗菌药物的敏感性。
    方法:在2022年3月至9月之间,使用最低抑制浓度(MIC)测试了从66只具有UTI症状的狗和41只猫的尿液中培养的大肠杆菌分离株的耐药性。对氨苄青霉素进行了抗菌药物敏感性试验,氨苄西林/舒巴坦,头孢唑啉,头孢呋辛,氨曲南,庆大霉素,阿米卡星,粘菌素,甲氧苄啶/磺胺甲恶唑,环丙沙星,氯霉素和四环素。
    结果:据记载,氨苄青霉素耐药率最高(68%的狗,100%在猫中)和氨苄西林与舒巴坦(59%在狗中,54%的猫)。大肠杆菌最常见的抗生素耐药模式是单独的氨苄西林(12个分离株,猫中29.3%)和β-内酰胺,包括氨曲南(14个分离株,狗中的21.2%)。
    结论:对氨曲南的高耐药性(61%和32%的狗和猫分离株,分别),其他β-内酰胺,和氟喹诺酮类药物应引起警报,因为动物共患病的潜力和抗生素抗性微生物在动物和人类之间的交叉传播。
    OBJECTIVE: Escherichia coli is one of the most common bacteria isolated from urine samples collected from dogs and cats with urinary tract infection (UTI). Uncomplicated UTIs in dogs and cats can be treated with short courses of first-line antimicrobial drugs, e.g. amoxicillin, amoxicillin with clavulanic acid, or trimethoprim/sulfonamide. Recurrent or complicated UTIs often require long-term treatment with broad-spectrum antibiotics. However, the choice of drug should be based on antimicrobial susceptibility.
    METHODS: Between March - September 2022, E. coli isolates cultured from the urine of 66 dogs and 41 cats with UTI symptoms were tested for antimicrobial resistance by using Minimum Inhibitory Concentration (MIC). Antimicrobial susceptibility was tested for ampicillin, ampicillin/sulbactam, cefazolin, cefuroxime, aztreonam, gentamycin, amikacin, colistin, trimethoprim/sulfamethoxazole, ciprofloxacin, chloramphenicol and tetracycline.
    RESULTS: The highest prevalence of resistance was documented for ampicillin (68% in dogs, 100% in cats) and ampicillin with sulbactam (59% in dogs, 54% in cats). The most common antimicrobial resistance patterns of E. coli were ampicillin alone (12 isolates, 29.3% in cats) and beta-lactams, including aztreonam (14 isolates, 21.2% in dogs).
    CONCLUSIONS: High resistance to aztreonam (61% and 32% of isolates from dogs and cats, respectively), other beta-lactams, and fluoroquinolones should cause be alarm due to zoonotic potential and cross-transmission of antimicrobial-resistant microorganisms between animals and humans.
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  • 文章类型: Journal Article
    肺毛细血管血管瘤病(PCH)是一种特发性疾病,肺组织中的小毛细血管样血管异常增殖,这可能导致严重的PH。兽医文献中描述的PCH只有几例:狗27例,猫2例。在兽医学中,PH主要被认为是左心衰竭的结果,是毛细血管后PH向毛细血管前形式的进展。PCH主要被描述为原发性疾病,但是,具有很高的肺水肿可能性的毛细血管后PH的耐药性引起了人们的猜测,即PCH可能是左心疾病的继发性畸形。
    在PH发展的背景下,发现与左侧和右侧心脏病之间转移相关的特征。
    回顾性分析具有PCH(sPCH)组织学标记的猫和狗的材料与右心衰竭(RHF)的材料。
    具有PCH的组织学和免疫组织化学标记的动物先前有患有左心容量超负荷的疾病史。两组之间的X线片和大体病理没有差异。组织学上,在RHF中可以发现肺纤维化和动脉病;在sPCH中,肺泡间隔中的毛细血管重复和周围结构中的奇异增殖。
    PCH可能是由于容量超负荷引起的血管重塑的次要模式。
    UNASSIGNED: Pulmonary capillary hemangiomatosis (PCH) is an idiopathic disease with the anomalous proliferation of a small capillary-like vessel in the pulmonary tissue, which can lead to a severe form of PH. There are only several cases of PCH described in veterinary literature: 27 cases in dogs and 2 cases in cats. In veterinary medicine, PH is mostly recognized as a consequence of left heart failure as a progression of the postcapillary PH to the precapillary form. PCH is mostly described as a primary disease, but resistant postcapillary PH with the high possibility of pulmonary edema raises speculation that PCH could be a secondary malformation to the left heart disease.
    UNASSIGNED: Discover the features associated with the shift between left- and right-sided heart disease in the context of PH development.
    UNASSIGNED: Retrospective analysis of materials from cats and dogs with histological markers of PCH (sPCH) versus those with right heart failure (RHF).
    UNASSIGNED: Animals with histological and immunohistochemistry markers of PCH had a previous history of disease with left heart volume overload. There were no differences between the groups in radiography and gross pathology. Histologically, pulmonary fibrosis and arteriopathy could be found in RHF; in sPCH-a duplication of capillaries in alveolar septa and bizarre proliferation in surrounding structures.
    UNASSIGNED: PCH could be a secondary pattern of vascular remodeling due to volume overload.
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  • 文章类型: Journal Article
    假性血小板减少症是在通过自动化机器分析猫血小板(PLT)计数时通常获得的假阴性结果。它与乙二胺四乙酸(EDTA)有关,一种广泛用于采血管的抗凝剂,导致EDTA依赖性假性血小板减少症(EDTA-PTCP)。
    为了研究用卡那霉素处理是否增加了使用EDTA-PTCP收集的猫科动物血液样本中PLT聚集的量。
    使用EDTA管获得31个血液样品。使用自动化的MindrayBC-5000Vet分析全血细胞计数。进行手动细胞计数和稀薄血液涂片以估计红细胞的数量,白细胞,和PLT以及评估PLT聚集的严重程度评分,分别。在EDTA管中进行预处理的那些和用卡那霉素处理的那些之间进行比较。
    在用卡那霉素处理之前和之后,样品的平均PLT计数显着不同,两者都是自动的(156.6±76.4vs.260.3±115.5;p<0.001)和手动(168.5±92.1与262.8±119.6;p<0.001)读数,95%置信区间为0.19(0.022-0.365)。
    这项研究表明,在临床实验室实践中,应将卡那霉素与EDTA-PTCP一起添加到猫科动物的血液样本中。
    UNASSIGNED: Pseudothrombocytopenia is a commonly obtained false negative result when analyzing feline platelet (PLT) count by an automated machine. It is related to ethylenediamine tetra-acetic acid (EDTA), a widely utilized anticoagulant in blood collection tubes, resulting in EDTA-dependent pseudothrombocytopenia (EDTA-PTCP).
    UNASSIGNED: To investigate whether treated with kanamycin enhanced the quantity of PLT aggregations in feline blood specimens collected using EDTA-PTCP.
    UNASSIGNED: Thirty-one blood samples were obtained using EDTA tubes. The complete blood count was analyzed using an automated Mindray BC-5000Vet. Both Manual cell counts and thin blood smears were performed to estimate the amount of red blood cell, white blood cell, and PLTs as well as to evaluate the severity scores of PLT clumping, respectively. Comparisons were made between those pre-treated and those treated with kanamycin in the EDTA tube.
    UNASSIGNED: There were significantly different mean PLT counts in the samples before and after they were treated with kanamycin, both on automated (156.6 ± 76.4 vs. 260.3 ± 115.5; p < 0.001) and manual (168.5 ± 92.1 vs. 262.8 ± 119.6; p < 0.001) readings, with a 95% confidence interval of 0.19 (0.022-0.365).
    UNASSIGNED: This study suggests that in clinical laboratory practice, kanamycin should be added to feline blood specimens with EDTA-PTCP.
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  • 文章类型: Journal Article
    背景:数字成像与基于深度学习的计算图像分析相结合是医学诊断中不断增长的领域,包括寄生虫学,其中已经开发了许多自动分析设备,并可用于临床实践。
    方法:ParasightAll-in-one(AIO)的性能,第二代设备,通过将其与公认的研究方法(mini-FLOTAC)和另一种市售测试(Imagyst)进行比较来进行评估。通过所有三种方法对59个犬和猫感染的粪便标本进行了定量分析。由于一些样本对一种以上的寄生虫呈阳性,该数据集包括48个Ancylostomaspp阳性标本。,13为弓形虫属。和23为毛虫属。
    结果:ParasightAIO计数的大小与mini-FLOTAC的大小相关,但与Imagyst的大小无关。ParasightAIO计数的Ancylostomaspp卵增加了约3.5倍。和Trichurisspp.弓形虫属的卵子增加了4.6倍。而不是迷你FLOTAC,并计算了27.9-,这些相同的卵子比Imagyst多17.1倍和10.2倍,分别。这些差异转化为在低卵计数水平(<50卵/g)下测试灵敏度之间的差异,与ParasightAIO>mini-FLOTAC>Imagyst。在更高的卵数下,ParasightAIO和mini-FLOTAC以可比的精度进行(明显高于Imagyst),而在较低计数(>30个鸡蛋/g)时,Parasight比mini-FLOTAC和Imagyst更精确,而后两种方法没有显着差异。
    结论:一般来说,ParasightAIO分析比mini-FLOTAC和Imagyst更精确,更灵敏,并且与mini-FLOTAC定量相关。虽然ParasightAIO生产的每克鸡蛋的原始计数低于mini-FLOTAC,这些可以使用从这些相关性产生的数据进行校正。
    BACKGROUND:  Digital imaging combined with deep-learning-based computational image analysis is a growing area in medical diagnostics, including parasitology, where a number of automated analytical devices have been developed and are available for use in clinical practice.
    METHODS: The performance of Parasight All-in-One (AIO), a second-generation device, was evaluated by comparing it to a well-accepted research method (mini-FLOTAC) and to another commercially available test (Imagyst). Fifty-nine canine and feline infected fecal specimens were quantitatively analyzed by all three methods. Since some samples were positive for more than one parasite, the dataset consisted of 48 specimens positive for Ancylostoma spp., 13 for Toxocara spp. and 23 for Trichuris spp.
    RESULTS: The magnitude of Parasight AIO counts correlated well with those of mini-FLOTAC but not with those of Imagyst. Parasight AIO counted approximately 3.5-fold more ova of Ancylostoma spp. and Trichuris spp. and 4.6-fold more ova of Toxocara spp. than the mini-FLOTAC, and counted 27.9-, 17.1- and 10.2-fold more of these same ova than Imagyst, respectively. These differences translated into differences between the test sensitivities at low egg count levels (< 50 eggs/g), with Parasight AIO > mini-FLOTAC > Imagyst. At higher egg counts Parasight AIO and mini-FLOTAC performed with comparable precision (which was significantly higher that than Imagyst), whereas at lower counts (> 30 eggs/g) Parasight was more precise than both mini-FLOTAC and Imagyst, while the latter two methods did not significantly differ from each other.
    CONCLUSIONS: In general, Parasight AIO analyses were both more precise and sensitive than mini-FLOTAC and Imagyst and quantitatively correlated well with mini-FLOTAC. While Parasight AIO produced lower raw counts in eggs-per-gram than mini-FLOTAC, these could be corrected using the data generated from these correlations.
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  • 文章类型: Journal Article
    立克次体在世界范围内发生,立克次体病在世界几个地区被认为是一种新兴的感染。蜱是人类和家畜中致病性立克次体物种的水库宿主。大多数致病性立克次体属斑点热组(SFG)。本研究旨在识别和诊断蜱动物,并调查立克次体的患病率。在克尔曼省农村地区的家畜和狗身上采集的蜱虫中,伊朗东南部。在这项研究中,鉴定了tick虫物种(动物),并使用实时PCR检测了来自两个属和物种的2100个tick虫(350个合并样品),其中包括Rhipicephaluslinnaei(1128)和Hyalommadeteriitum(972)。在24.9%(95CI20.28-29.52)的合并样品中观察到立克次体属的存在。测序和系统发育分析显示存在立克次体aeschlimannii(48.98%),以色列立克次体(28.57%),Sibirica立克次体(20.41%),阳性样本中的立克次体(2.04%)。结果表明,县变量与以下变量之间存在显着关联:tickspp。(p<0.001),蜱(p<0.001)和立克次体属感染。(p<0.001)。此外,蜱物种和宿主动物(狗和家畜)之间存在显著关联(p<0.001),蜱的立克次体感染(p<0.001),和立克次体。(p<0.001)。这项研究表明立克次体的患病率很高。(SFG)在克尔曼省农村地区的家畜和狗中。应告知卫生系统立克次体病的可能性以及这些地区立克次体的流行物种。
    Rickettsia occurs worldwide and rickettsiosis is recognized as an emerging infection in several parts of the world. Ticks are reservoir hosts for pathogenic Rickettsia species in humans and domestic animals. Most pathogenic Rickettsia species belong to the spotted Fever Group (SFG). This study aimed to identify and diagnose tick fauna and investigate the prevalence of Rickettsia spp. in ticks collected from domestic animals and dogs in the rural regions of Kerman Province, Southeast Iran. In this study, tick species (fauna) were identified and 2100 ticks (350 pooled samples) from two genera and species including Rhipicephalus linnaei (1128) and Hyalomma deteritum (972) were tested to detect Rickettsia genus using Real-time PCR. The presence of the Rickettsia genus was observed in 24.9% (95%CI 20.28-29.52) of the pooled samples. Sequencing and phylogenetic analyses revealed the presence of Rickettsia aeschlimannii (48.98%), Rickettsia conorii israelensis (28.57%), Rickettsia sibirica (20.41%), and Rickettsia helvetica (2.04%) in the positive samples. The results showed a significant association between county variables and the following variables: tick spp. (p < 0.001), Rickettsia genus infection in ticks (p < 0.001) and Rickettsia spp. (p < 0.001). In addition, there was a significant association between tick species and host animals (dogs and domestic animals) (p < 0.001), Rickettsia spp infection in ticks (p < 0.001), and Rickettsia spp. (p < 0.001). This study indicates a high prevalence of Rickettsia spp. (SFG) in ticks of domestic animals and dogs in rural areas of Kerman Province. The health system should be informed of the possibility of rickettsiosis and the circulating species of Rickettsia in these areas.
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  • 文章类型: Journal Article
    逆转录病毒的包膜糖蛋白(Env),例如猫白血病病毒(FeLV),是中和体液反应的主要目标,因此,一个有前途的候选疫苗,尽管据报道其免疫原性差。在融合前构象中掺入稳定来自其他病毒的类似蛋白的突变(例如,艾滋病毒感染,SARS-CoV-2S,或RSVF糖蛋白)提高了它们诱导中和保护性免疫应答的能力。因此,我们已经稳定了FeLVEnv蛋白,该策略基于先前用于生成可溶性HIVEnv三聚体的二硫键和Ile/Pro突变(SOSIP)的掺入。我们已经将这种SOSIP-FeLVEnv表征为其可溶形式,并作为高密度存在于基于FeLVGag的VLP表面上的跨膜蛋白。此外,我们已经在C57BL/6小鼠的DNA免疫试验中测试了其免疫原性。在SOSIP-FeLV可溶性蛋白免疫的动物中检测到低的抗FeLVEnv应答;然而,在用基于SOSIP-FeLVGag的VLP免疫的动物中意外地没有检测到应答。相比之下,在用缺乏SOSIP-FeLVEnv的对照GagVLP免疫的动物中观察到针对FeLVGag的高体液应答,而当VLP掺入SOSIP-FeLVEnv时,这种反应明显受损。我们的数据表明,FeLVEnv可以稳定为可溶性蛋白,并且可以在高密度VLP中表达。然而,当配制成DNA疫苗时,SOSIP-FeLVEnv仍然缺乏免疫原性,开发有效的FeLV疫苗必须克服的限制。
    The envelope glycoprotein (Env) of retroviruses, such as the Feline leukemia virus (FeLV), is the main target of neutralizing humoral response, and therefore, a promising vaccine candidate, despite its reported poor immunogenicity. The incorporation of mutations that stabilize analogous proteins from other viruses in their prefusion conformation (e.g., HIV Env, SARS-CoV-2 S, or RSV F glycoproteins) has improved their capability to induce neutralizing protective immune responses. Therefore, we have stabilized the FeLV Env protein following a strategy based on the incorporation of a disulfide bond and an Ile/Pro mutation (SOSIP) previously used to generate soluble HIV Env trimers. We have characterized this SOSIP-FeLV Env in its soluble form and as a transmembrane protein present at high density on the surface of FeLV Gag-based VLPs. Furthermore, we have tested its immunogenicity in DNA-immunization assays in C57BL/6 mice. Low anti-FeLV Env responses were detected in SOSIP-FeLV soluble protein-immunized animals; however, unexpectedly no responses were detected in the animals immunized with SOSIP-FeLV Gag-based VLPs. In contrast, high humoral response against FeLV Gag was observed in the animals immunized with control Gag VLPs lacking SOSIP-FeLV Env, while this response was significantly impaired when the VLPs incorporated SOSIP-FeLV Env. Our data suggest that FeLV Env can be stabilized as a soluble protein and can be expressed in high-density VLPs. However, when formulated as a DNA vaccine, SOSIP-FeLV Env remains poorly immunogenic, a limitation that must be overcome to develop an effective FeLV vaccine.
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