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Isolation, Identification, and Prevalence of Oral Mycoplasmas in North American Pinnipeds. 北美鳍足类口腔支原体的分离、鉴定和流行。
IF 1.2 4区 农林科学 Q3 VETERINARY SCIENCES Pub Date : 2026-02-09 DOI: 10.7589/JWD-D-25-00088
Lena N Measures

Seal finger is a zoonotic infection that humans acquire from pinnipeds during dissection or handling, or from trauma. This infection is not uncommon among Canadian biologists working with wild pinnipeds. The etiological agent is believed to be mycoplasma bacteria. No comprehensive data on prevalence of oral mycoplasmas in pinnipeds are available. To investigate whether pinnipeds carry oral mycoplasmas, wild (n=148) and captive (n=14) phocids in eastern Canada and pinnipeds (n=38) in a rehabilitation center in California, USA, were sampled from 1996 to 1998. Mycoplasmas were cultured, isolated, and identified using indirect fluorescent antibody tests in a dedicated mycoplasma laboratory, and prevalence by sex, age, age class, and species of seal was determined. A total of 23l mycoplasma isolates were obtained from sampled pinnipeds. In Canadian phocids, Mycoplasma phocicerebrale, Mycoplasma phocae, and Mycoplasma phocirhinis were isolated and identified from wild gray (Halichoerus grypus) and Atlantic harbor (Phoca vitulina concolor) seals, while M. phocicerebrale was isolated from wild hooded (Cystophora cristata) and harp (Pagophilus groenlandicus) seals and from captive gray and harbor seals, including new host and geographic reports. In wild seals, prevalence of M. phocicerebrale was 78% (31/40) in gray, 70% (32/46) in hooded, 44% (11/25) in harbor, and 22% (8/37) in harp seals. In Californian pinnipeds, M. phocicerebrale and M. phocirhinis were isolated and identified from California sea lions (Zalophus californianus), northern elephant (Mirounga angustirostris) seals, and Pacific harbor (Phoca vitulina richardii) seals, also new host and geographic reports. Unidentified Mycoplasma and Ureaplasma spp. were also isolated in wild and captive Canadian and Californian pinnipeds. These new data on the prevalence and identification of oral mycoplasmas in sampled pinnipeds indicate that personnel working with these animals in the field or in captive situations should exercise caution.

海豹指是一种人畜共患感染,人类在解剖或处理鳍足动物或外伤时获得。这种感染在研究野生鳍足动物的加拿大生物学家中并不罕见。病原被认为是支原体细菌。没有关于鳍足类动物口腔支原体流行率的综合数据。为了调查鳍足类动物是否携带口腔支原体,1996 - 1998年在加拿大东部采集野生(148只)和圈养(14只)鳍足类动物样本,并在美国加利福尼亚州某康复中心采集鳍足类动物样本(38只)。支原体在专门的支原体实验室中培养、分离并使用间接荧光抗体试验进行鉴定,并按性别、年龄、年龄类别和海豹种类确定患病率。从鳍足类中分离出支原体23株。在加拿大海豹中,从野生灰海豹(Halichoerus grypus)和大西洋港海豹(Phoca vitulina concolor)中分离和鉴定出了光脑支原体、phocicerebrale支原体和phocirhinis支原体,而从野生冠海豹(Cystophora cristata)和格陵兰海豹(Pagophilus groenlandicus)以及圈养灰海豹和港海豹中分离出了phocicerebrale支原体,包括新的宿主和地理报告。在野生海豹中,灰海豹、帽海豹、港海豹和竖琴海豹中,光脑微囊藻的患病率分别为78%(31/40)、70%(32/46)、44%(11/25)和22%(8/37)。在加利福尼亚鳍足动物中,从加利福尼亚海狮(Zalophus californianus)、北象海豹(miounga angustirostris)和太平洋港海豹(Phoca vitulina richardii)中分离鉴定出了M. phocicerebrale和M. phocirhinis,这也是新的宿主和地理报告。在野生和圈养的加拿大和加利福尼亚鳍足类动物中也分离到未鉴定的支原体和脲原体。这些关于在取样的鳍足类动物中流行和鉴定口腔支原体的新数据表明,在野外或圈养环境中与这些动物打交道的人员应谨慎行事。
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引用次数: 0
Hematologic Parameters of Free-Ranging Screaming Hairy Armadillos (Chaetophractus vellerosus). 散养尖叫毛犰狳的血液学参数。
IF 1.2 4区 农林科学 Q3 VETERINARY SCIENCES Pub Date : 2026-02-09 DOI: 10.7589/JWD-D-24-00163
Noralí Pagnutti, Luisina Fux, Ailín Carballo Luchetti, Carolina Plez, Mariané B Mañez, Clara Vercellini, Josefina Lacunza, Cecilia M Ezquiaga, Agustín M Abba

Hematologic parameters from free-ranging screaming hairy armadillos, Chaetophractus vellerosus, were determined by hemocytometry and blood smear review. Packed cell volume was higher and basophil and eosinophil counts lower than those reported for this species in captivity. These findings will serve as a reference for future research.

采用血细胞计数法和血液涂片法对散养尖叫毛犰狳(Chaetophractus vellerosus)的血液学参数进行了测定。堆积细胞体积较高,嗜碱性粒细胞和嗜酸性粒细胞计数低于圈养该物种的报道。这些发现将为今后的研究提供参考。
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引用次数: 0
Confirmation of Chelonid Alphaherpesvirus 5 (Scutavirus Chelonidalpha5) Associated with Fibropapillomatosis in Green and Kemp's Ridley Sea Turtles from Mississippi, USA. 美国密西西比州绿海龟和肯普氏雷德利海龟纤维乳头状瘤病与龟类甲疱疹病毒5 (Chelonidalpha5)相关性的证实
IF 1.2 4区 农林科学 Q3 VETERINARY SCIENCES Pub Date : 2026-02-09 DOI: 10.7589/JWD-D-24-00217
Sarah A Rubelowsky, Justin M Stilwell, Caroline Betbeze, Samantha Hughes, Debra Moore, Lyndsey Howell, Natalie K Stilwell

We diagnosed and molecularly confirmed the presence of fibropapillomatosis and chelonid alphaherpesvirus 5 infection in one Kemp's ridley sea turtle (Lepidochelys kempii) and three green sea turtles (Chelonia mydas) stranded on the Mississippi, USA, coast, expanding the known range of this disease in sea turtles in the Gulf of Mexico.

我们诊断并分子证实了在美国密西西比海岸搁浅的一只肯普雷氏海龟(Lepidochelys kempii)和三只绿海龟(Chelonia mydas)中存在纤维乳头状瘤病和龟类α疱疹病毒5感染,扩大了墨西哥湾海龟中该疾病的已知范围。
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引用次数: 0
Radiographic Study of Fractures and Luxations in Raptors Admitted at Two Wildlife Centers, Located in Brazil and the USA. 位于巴西和美国的两个野生动物中心对迅猛龙骨折和脱位的放射学研究。
IF 1.2 4区 农林科学 Q3 VETERINARY SCIENCES Pub Date : 2026-02-09 DOI: 10.7589/JWD-D-24-00211
Heloísa Coppini de Lima, Sheila Canevese Rahal, Krystal M T Woo, Peter J Flowers, Tamires Goneli Wichert Teodoro, Jeana Pereira da Silva, Maria Jaqueline Mamprim, Joanne R Paul-Murphy, Miriam Harumi Tsunemi, Anneke Moresco

A radiographic analysis of fractures and/or luxations in raptors was conducted across two wildlife centers, one in Brazil (Center for Medicine and Research in Wild Animals [CEMPAS]; n=106 raptors) and the other in the USA (Lindsay Wildlife Experience [LWE]; n=310 raptors), through a data study spanning 7 yr. There were differences between the two centers in which order was most affected. At CEMPAS, Strigiformes accounted for 48.1% of the cases, but at LWE Accipitriformes were most affected (59.4% of cases). The axial skeleton was least affected at both centers, with two cases at CEMPAS and three cases at LWE. Most fractures included a single bone in the appendicular skeleton, accounting for 57.7% at CEMPAS and 61.9% at LWE. In the thoracic limb, the humerus was the most fractured bone at CEMPAS (44/108), whereas the ulna was most commonly fractured at LWE (116/307). In the pelvic limb, the tibiotarsus was the most fractured bone at both centers (23/35 at CEMPAS and 27/80 at LWE). Closed fractures were more frequent in the thoracic and pelvic limbs at both centers. Cases of luxation or subluxation without fractures were less common, with 6.7% at CEMPAS and 7.2% at LWE. The types of fractures and/or luxations identified in this study exhibited several similarities between centers that may be useful in understanding these lesions in raptors and establishing future treatment protocols.

通过7年的数据研究,在两个野生动物中心对猛禽的骨折和/或脱位进行了放射学分析,一个在巴西(野生动物医学和研究中心[CEMPAS]; n=106只猛禽),另一个在美国(林赛野生动物体验中心[LWE]; n=310只猛禽)。两个中心之间的差异是,顺序受影响最大。在CEMPAS,返形虫占48.1%,但在LWE,反形虫受影响最大(59.4%)。两个中心的轴向骨骼受影响最小,CEMPAS有2例,LWE有3例。大多数骨折包括附肢骨骼中的一根骨头,在CEMPAS和LWE中分别占57.7%和61.9%。在胸肢中,肱骨在CEMPAS骨折最多(44/108),而尺骨在LWE骨折最常见(116/307)。在骨盆肢体中,胫跗骨是两个中心骨折最多的骨(CEMPAS为23/35,LWE为27/80)。闭合性骨折在两个中心的胸肢和盆腔肢中更为常见。脱位或半脱位无骨折的病例较少见,CEMPAS为6.7%,LWE为7.2%。本研究中确定的骨折和/或脱位类型在中心之间显示出一些相似之处,这可能有助于了解迅猛龙的这些病变,并建立未来的治疗方案。
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引用次数: 0
Surveillance of Anticoagulant Rodenticides in Free-Roaming Outdoor Cats (Felis catus) in Chicago, Illinois, USA. 美国伊利诺斯州芝加哥市流浪猫(Felis catus)体内抗凝血灭鼠剂的监测。
IF 1.2 4区 农林科学 Q3 VETERINARY SCIENCES Pub Date : 2026-02-09 DOI: 10.7589/JWD-D-25-00092
Maureen H Murray, Jacqueline Y Buckley, Seth Magle

Rats are managed using anticoagulant rodenticides (ARs) in cities worldwide, but these compounds cause morbidity and mortality in nontarget wildlife. Several cities also use free-roaming outdoor cat (Felis catus) colonies to help reduce rats, but there have been no investigations of how often these cats are exposed to ARs. In this study, our goal was to assess how often free-roaming domestic cats are exposed to ARs to 1) understand health risks for urban predators by using free-roaming cats as a proxy and 2) evaluate the health risks associated with using cats as rodent control. We tested blood samples from 57 cats that were part of trap-neuter-release cat colonies in Chicago, Illinois, USA. We found that 7% (4/57) of cats tested positive for one type of rodenticide at trace amounts. Free-roaming cats can therefore be exposed to ARs, although the prevalence and concentrations were lower than in wild carnivores. Future research is needed to understand how often cats may be exposed to ARs by killing or consuming rats and any health impacts. Stronger rodenticide regulations are needed to reduce health risks for urban predators, and owned cats should be kept indoors to minimize the risk of rodenticide exposure.

在世界各地的城市中,老鼠使用抗凝血灭鼠剂(ARs)进行管理,但这些化合物会导致非目标野生动物的发病率和死亡率。一些城市还使用自由漫游的户外猫群(Felis catus)来帮助减少老鼠,但没有调查这些猫暴露于ARs的频率。在这项研究中,我们的目标是评估自由漫游的家猫暴露于ARs的频率,以1)通过将自由漫游的猫作为代理来了解城市捕食者的健康风险;2)评估与猫作为啮齿动物控制相关的健康风险。我们测试了57只猫的血液样本,这些猫是美国伊利诺伊州芝加哥市捕获的绝育猫群的一部分。我们发现7%(4/57)的猫对微量的一种灭鼠剂检测呈阳性。因此,自由漫游的猫可能暴露于ARs,尽管其发病率和浓度低于野生食肉动物。未来的研究需要了解猫通过杀死或食用老鼠暴露于ARs的频率以及对健康的影响。需要制定更强有力的灭鼠剂法规,以减少城市捕食者的健康风险,并应将养猫关在室内,以尽量减少接触灭鼠剂的风险。
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引用次数: 0
Giant Liver Fluke (Fascioloides magna) in Free-Ranging Muskoxen (Ovibos moschatus) of Nunavik, Québec, Canada. 加拿大魁省努那维克散养麝牛(Ovibos moschatus)中巨型肝吸虫(大片形吸虫)。
IF 1.2 4区 农林科学 Q3 VETERINARY SCIENCES Pub Date : 2026-02-09 DOI: 10.7589/JWD-D-25-00047
Charlotte Nury, Laura Van Driessche, Ariane Santamaria-Bouvier, Manon Simard, Vincent Brodeur, Joëlle Taillon, Stéphane Lair

We describe hepatic infections by giant liver fluke (Fascioloides magna) in free-ranging muskoxen (Ovibos moschatus) in Nunavik, Québec, Canada. Whole livers and feces were collected from 51 muskoxen (37 males, 14 females) harvested from 2007 to 2011. Prevalence of F. magna was very high (48/51, 94%; 95% confidence interval, 84-99), with the intensity of infection reaching up to 40 adult trematodes per liver (median, 4 flukes). The estimated percentage of macroscopically altered liver, which was evaluated in serial 1-cm-thick slices, ranged from 0.1% to 57.3% and was positively correlated with the number of parasites detected. Animals harvested near Tasiujaq had higher mean fluke intensity (7.2) than those near Kuujjuaq (2.6) in Nunavik, Québec, Canada. Despite the extent of the hepatic alterations in some individuals, no association was found between intensity of infection or liver alteration and the examined nutritional condition indices (depth of back fat, femur marrow fat percentage, and kidney fat index). Field observations suggest that infections by F. magna did not have a significant clinical impact on sampled muskoxen. The mean fluke intensity and the percentage of altered liver did not vary between sex and age. Fecal shedding of F. magna eggs was documented in some individuals, indicating that muskoxen could be characterized as a definitive host. Further research is needed to define the reservoir species and characterize the life cycle of this parasite in the Canadian Arctic.

我们描述了由巨型肝吸虫(大片形吸虫)在自由放养的麝牛(Ovibos moschatus)在Nunavik, quacimbec,加拿大肝脏感染。收集了2007 - 2011年收获的51头麝牛(公37头,母14头)的全肝和粪便。magna F.的患病率非常高(48/51,94%;95%可信区间为84-99),感染强度高达每肝40个成年吸虫(中位数为4个吸虫)。在一系列1厘米厚的切片中评估的肝脏宏观改变的估计百分比在0.1%至57.3%之间,与检测到的寄生虫数量呈正相关。在加拿大魁省努纳维克,Tasiujaq附近收获的动物平均吸虫强度(7.2)高于Kuujjuaq(2.6)。尽管在某些个体中存在肝脏改变的程度,但未发现感染强度或肝脏改变与所检查的营养状况指标(背部脂肪深度、股骨骨髓脂肪百分比和肾脏脂肪指数)之间存在关联。实地观察表明,感染F. magna对样本麝牛没有显著的临床影响。平均吸虫强度和肝脏改变的百分比在性别和年龄之间没有变化。在一些个体中记录了F. magna卵的粪便脱落,表明麝牛可以被定性为最终宿主。需要进一步的研究来确定储层物种,并描述这种寄生虫在加拿大北极的生命周期。
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引用次数: 0
Exploratory Study of the Effect of Postmortem Interval on the Biochemical Quality of Wild Boar (Sus scrofa) Blood. 宰后时间对野猪血液生化品质影响的探索性研究。
IF 1.2 4区 农林科学 Q3 VETERINARY SCIENCES Pub Date : 2026-02-09 DOI: 10.7589/JWD-D-25-00019
Sylvain Larrat, Benoît Rannou, Léa Bonin, Karin Lemberger, Julie Tucoulet, Ariane Payne, Clément Calenge, Anouk Decors

Diagnosing the cause of peracute death in wildlife is challenging, particularly when necropsy, histopathology, and ancillary testing yield nonspecific results. Biochemistry could provide pathophysiologic information on the death process that is unobtainable with morphologic methods. To evaluate the impact of postmortem delay on the quality of blood biochemical analysis, blood samples were collected from 20 wild boars (Sus scrofa) hunted in January 2018 in France. The body condition, age, and sex of each boar were recorded. Each animal was sampled once between the time of death and 6 h postmortem, with six boars sampled within the first hour, three boars sampled at each hour thereafter (hours 2-5), and two boars sampled at 6 h postmortem. Samples were promptly sent to the laboratory, centrifuged, and assessed for hemolysis before measurement of biochemical parameters using a wet chemistry analyzer. The first component from a principal component analysis was used as a quality index of the biochemical composition of the blood. This index strongly correlated positively with sodium and chloride and negatively with total proteins, alanine aminotransferase, fructosamine, and potassium. A segmented regression analysis indicated stability of blood quality for 2 h after death, followed by a linear decrease. Practically, blood samples drawn within 2 h after death maintained overall quality. This exploratory study should be expanded with evaluations of changes of individual metabolites over time.

诊断野生动物的过急性死亡原因具有挑战性,特别是当尸检、组织病理学和辅助检测产生非特异性结果时。生物化学可以提供形态学方法无法获得的死亡过程的病理生理学信息。为了评估死后延迟对血液生化分析质量的影响,研究人员采集了2018年1月在法国猎杀的20只野猪(Sus scrofa)的血液样本。记录每头公猪的身体状况、年龄和性别。每头猪在死亡至死后6小时取样一次,头1小时取样6头,此后(2-5小时)每小时取样3头,死后6小时取样2头。样品立即送到实验室,离心,并评估溶血,然后使用湿化学分析仪测量生化参数。主成分分析的第一个成分被用作血液生化成分的质量指标。该指标与钠、氯呈显著正相关,与总蛋白、丙氨酸转氨酶、果糖胺、钾呈显著负相关。分段回归分析表明,血液质量在死亡后2小时内保持稳定,随后呈线性下降。实际上,在死亡后2小时内抽取的血液样本保持了总体质量。这项探索性研究应该随着个体代谢物随时间变化的评估而扩大。
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引用次数: 0
Elevated Exposure to Influenza A Viruses in Wild Pigs (Sus scrofa), Texas, USA. 美国德克萨斯州野猪(Sus scrofa)甲型流感病毒暴露量升高。
IF 1.2 4区 农林科学 Q3 VETERINARY SCIENCES Pub Date : 2026-02-09 DOI: 10.7589/JWD-D-25-00031
Kayleigh Chalkowski, Ryan S Miller, Jeffrey C Chandler, Alessandra Campos, Bailey Arruda, Amy L Baker, Tavis K Anderson, Jason Klemm, Vienna R Brown, Kurt C Verauteren, Nathan P Snow, Kim M Pepin

Detections of highly pathogenic avian influenza viruses in mammals have increased globally, underscoring the need to assess influenza A virus (IAV) exposure in species that bridge wildlife, livestock, and human interfaces. We collected serum from 396 wild pigs (Sus scrofa, hybrids of wild boar and domestic swine) between October 2022 and May 2023 on a cattle ranch in north central Texas, USA, to estimate IAV seroprevalence and identify ecologic correlates of exposure. Using commercial blocking ELISAs and hemagglutination inhibition assays, we report high recent exposure of wild pigs to IAVs (seroprevalence 40%, n=396). Seropositivity was highest in individuals that were female, >2 yr old, sampled during spring (compared with fall or winter), and/or sampled <1.5 km from surface water. Almost half (46.3%, 57/123) of the samples were positive for at least two hemagglutinin clade representatives, and 13.8% (17/123) had titers against both an H1 and an H3 strain. These results indicate a hotspot of IAV circulation in wild pigs in north central Texas and highlight elevated exposure near water bodies, where wild pigs may contact wild waterfowl. Enhanced surveillance of wild pigs at the wildlife-livestock-waterbird interface is warranted to better understand ecologic and epidemiologic pathways for IAV transmission and reassortment.

在全球范围内,在哺乳动物中检测到的高致病性禽流感病毒有所增加,这突出表明有必要评估连接野生动物、牲畜和人类界面的物种中甲型流感病毒(IAV)暴露情况。我们于2022年10月至2023年5月在美国德克萨斯州中北部的一个养牛场收集了396头野猪(Sus scrofa,野猪和家猪的杂交品种)的血清,以估计IAV血清患病率并确定暴露的生态相关性。使用商业阻断elisa和血凝抑制试验,我们报告了最近野猪暴露于iav的高水平(血清阳性率40%,n=396)。在春季(与秋季或冬季相比)和/或春季取样的女性,20至20岁的个体血清阳性最高
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引用次数: 0
Occurrence of Dystocia among Free-Ranging Giraffes (Giraffa spp.) in Kenya. 肯尼亚自由放养长颈鹿难产的发生。
IF 1.2 4区 农林科学 Q3 VETERINARY SCIENCES Pub Date : 2026-02-09 DOI: 10.7589/JWD-D-24-00105
Francis Gakuya, David Ndeereh, Isaac Lekolool, Ian Muchina, James Akoko, Richard Kock

Dystocia, a complication that occurs at parturition, either due to fetal or maternal factors, is uncommon in wild ungulate species because of natural selection against predisposed individuals. Among giraffes, reports of dystocia are rare in free-ranging populations across Africa. This study reviewed occurrence and type of dystocia, and outcome of obstetrical interventions, in giraffes in Kenya during 1991-2023 and considered possible risk factors. Only one case of dystocia in a semicaptive and one case in a captive northern (Rothschild's) Rothschild's giraffe (Giraffa camelopardalis) had been reported in Kenya before 2012. Seventeen cases were recorded in free-ranging giraffes during 2012-23 in 10 different protected areas with variable range and forage restrictions, associated with fenced, partially fenced, or unfenced management systems. In the latter period, three giraffe species found in Kenya were all affected, with nine cases in Masai giraffes (Giraffa tippelskirchi) and four cases each in reticulated giraffes (Giraffa reticulata) and Rothschild's giraffes. Cases were attributed to fetal factors, including malposition that refers to the position of the fetus or to malpresentation that refers to parts of the body presenting on the pelvis. Potential maternal factors were not evaluated in any individuals. Based on dam survival, 75% success rate relieving dystocia, through obstetrical manipulation, was recorded, but none of the calves survived. We postulate that dystocia might be increasing in Kenya, although increased reports may simply reflect improving capacities to observe and intervene.

难产是一种发生在分娩时的并发症,由于胎儿或母体因素,在野生有蹄类动物中并不常见,因为自然选择不利于易感个体。在非洲自由放养的长颈鹿中,难产的报道很少见。本研究回顾了1991-2023年肯尼亚长颈鹿难产的发生和类型,以及产科干预的结果,并考虑了可能的危险因素。在2012年之前,肯尼亚只报告了一例半圈养的罗斯柴尔德长颈鹿难产和一例圈养的罗斯柴尔德长颈鹿难产。2012-23年期间,在10个不同的保护区中,在自由放养的长颈鹿中记录了17例病例,这些保护区的活动范围和饲料限制不同,与围栏、部分围栏或非围栏管理系统有关。在后一时期,在肯尼亚发现的三种长颈鹿都受到了影响,马赛长颈鹿(Giraffa tippelskirchi)有9例,网状长颈鹿(Giraffa reticulata)和罗斯柴尔德长颈鹿各有4例。病例归因于胎儿因素,包括胎儿的体位不正,或身体部位出现在骨盆上的体位不正。未对任何个体的潜在母性因素进行评估。根据成活率,记录75%的成功率缓解难产,通过产科操作,但没有小牛存活。我们假设难产可能在肯尼亚增加,尽管增加的报告可能仅仅反映了观察和干预能力的提高。
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引用次数: 0
Investigation of Sample Storage Conditions for Optimized Detection of Frog Virus 3 DNA in Chelonian Swab Samples. 龟拭子样品中检测青蛙病毒3型DNA的最佳保存条件研究。
IF 1.2 4区 农林科学 Q3 VETERINARY SCIENCES Pub Date : 2026-02-09 DOI: 10.7589/JWD-D-24-00165
Carly R Harkey, Laura A Adamovicz, Maris J Daleo, Amber L Simmons, Matthew C Allender

The collection and storage of swab samples for molecular diagnostics is a routine component of wildlife health surveillance. The suitability of different sample storage conditions for maximizing the recovery of pathogen DNA in most species has not been assessed; therefore, the aim of this study was to identify a preferred storage method for swabs collected for the detection of frog virus 3 (FV3), a significant chelonian pathogen. Sterile swabs were inoculated in triplicate with a plasmid containing known quantities of FV3 DNA from 100 to 107 copies. Swabs were then stored under one of the following four conditions: 1) dry frozen at -20 °C; 2) immersed in phosphate-buffered saline (PBS) and frozen at -20 °C; 3) immersed in a nucleic acid preservative (RNAlater) and frozen at -20 °C; and 4) immersed in 100% ethanol at ambient room temperature. Swabs remained undisturbed under designated storage conditions for 14 d, at which point DNA extraction and conventional and quantitative PCR for FV3 detection were performed. Conventional PCR amplified down to the lowest expected FV3 target copy number (10,000 copies/swab) for the dry-frozen and PBS-frozen treatment groups. Conventional PCR amplification was inconsistent for the ethanol and RNAlater treatment groups. Quantitative PCR on dry-frozen samples successfully amplified as low as 100 FV3 target copies/swab with a mean recovery of 90%, with all other storage methods amplifying only down to 10,000 copies/swab. Findings suggest there is improved detection of pathogen DNA for samples stored from collection to extraction under the dry-frozen method. Swab sample storage recommendations for future applications should be observed within the context of study-specific objectives and target pathogens. Furthermore, failure to detect fewer than 100 copies/swab of FV3 from any storage method may have clinically significant ramifications and suggests that the differences in DNA recovery based on extraction method should also be examined.

收集和储存用于分子诊断的拭子样本是野生动物健康监测的常规组成部分。在大多数物种中,不同的样品储存条件对最大限度地恢复病原体DNA的适用性尚未得到评估;因此,本研究的目的是确定用于检测蛙病毒3 (FV3)(一种重要的龟病原体)收集的拭子的首选储存方法。用含有已知数量的FV3 DNA的质粒接种无菌拭子一式三次,从10°到107拷贝。然后将拭子在以下四种条件之一下保存:1)在-20°C干冷冻;2)浸泡在磷酸盐缓冲盐水(PBS)中,-20℃冷冻;3)浸泡在核酸保鲜剂(RNAlater)中,-20℃冷冻;4)在室温下浸泡在100%乙醇中。拭子在指定的储存条件下保持原状14天,此时进行DNA提取和FV3检测的常规和定量PCR。对于干冷冻和pbs冷冻处理组,常规PCR扩增至最低预期FV3目标拷贝数(10,000拷贝/拭子)。乙醇和RNAlater处理组常规PCR扩增结果不一致。干冻样品上的定量PCR成功扩增到100个FV3靶拷贝/拭子,平均回收率为90%,而所有其他储存方法的扩增率仅为10,000个拷贝/拭子。研究结果表明,在干冷冻方法下,从收集到提取的样品中,病原体DNA的检测得到了改进。应在研究特定目标和目标病原体的背景下观察未来应用的拭子样本存储建议。此外,未能从任何存储方法中检测到少于100拷贝/拭子的FV3可能会产生重大的临床后果,这表明还应检查基于提取方法的DNA恢复差异。
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Journal of Wildlife Diseases
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