Pub Date : 2024-10-10DOI: 10.1016/j.vetpar.2024.110328
Roger I. Rodríguez-Vivas , Melina M. Ojeda-Chi , Iris Trinidad-Martinez , Gabriela J. Flota-Burgos , Nadia F. Ojeda-Robertos
Amblyomma cf. parvum (Ixodida: Ixodidae) is an ectoparasite of domestic animals and wildlife on the American continent. There is little information on the efficacy of acaricides for the control of this tick species in domestic animals. Here, we determined the toxicological response of A. cf. parvum to amitraz, coumaphos, and cypermethrin. Engorged females of A. cf. parvum were collected (in two periods, eight months apart) from dogs in Yucatan, Mexico. The larval immersion test was used on the progeny of adult female ticks to test for amitraz susceptibility, and the larval package test was used to test for coumaphos and cypermethrin susceptibility. Dose–mortality regressions, lethal concentrations (LC), 95 % confidence intervals (CI95 %), and slopes were estimated by probit analysis. The lethal concentrations to kill 50 % of the tick population for amitraz, coumaphos, and cypermethrin in the first period were 1 ppm (CI95 %: 1–3 ppm), 10 ppm (CI95 %: 9–11 ppm), and 10 ppm (CI95 %: 9–10 ppm), respectively; meanwhile in the second period were 2 ppm (CI95 %: 1–3 ppm), 8 ppm (CI95 %: 6–10 ppm), and 9 ppm (CI95 %: 8–10 ppm), respectively. In conclusion, amitraz, coumaphos, and cypermethrin demonstrated high larvicidal efficacy in in vitro bioassays against A. cf. parvum populations infesting dogs.
{"title":"First documentation of dose response of Amblyomma cf. parvum population infesting dogs to amitraz, coumaphos, and cypermethrin","authors":"Roger I. Rodríguez-Vivas , Melina M. Ojeda-Chi , Iris Trinidad-Martinez , Gabriela J. Flota-Burgos , Nadia F. Ojeda-Robertos","doi":"10.1016/j.vetpar.2024.110328","DOIUrl":"10.1016/j.vetpar.2024.110328","url":null,"abstract":"<div><div><em>Amblyomma</em> cf. <em>parvum</em> (Ixodida: Ixodidae) is an ectoparasite of domestic animals and wildlife on the American continent. There is little information on the efficacy of acaricides for the control of this tick species in domestic animals. Here, we determined the toxicological response of <em>A</em>. cf. <em>parvum</em> to amitraz, coumaphos, and cypermethrin. Engorged females of <em>A</em>. cf. <em>parvum</em> were collected (in two periods, eight months apart) from dogs in Yucatan, Mexico. The larval immersion test was used on the progeny of adult female ticks to test for amitraz susceptibility, and the larval package test was used to test for coumaphos and cypermethrin susceptibility. Dose–mortality regressions, lethal concentrations (LC), 95 % confidence intervals (CI95 %), and slopes were estimated by probit analysis. The lethal concentrations to kill 50 % of the tick population for amitraz, coumaphos, and cypermethrin in the first period were 1 ppm (CI95 %: 1–3 ppm), 10 ppm (CI95 %: 9–11 ppm), and 10 ppm (CI95 %: 9–10 ppm), respectively; meanwhile in the second period were 2 ppm (CI95 %: 1–3 ppm), 8 ppm (CI95 %: 6–10 ppm), and 9 ppm (CI95 %: 8–10 ppm), respectively. In conclusion, amitraz, coumaphos, and cypermethrin demonstrated high larvicidal efficacy in <em>in vitro</em> bioassays against <em>A</em>. cf. <em>parvum</em> populations infesting dogs.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"332 ","pages":"Article 110328"},"PeriodicalIF":2.0,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142444935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Surface hydrophobicity of organisms provides biological self-protection. The hydrophobicity of pest surface, acting as a main obstacle for the pest control, can lead to low utilization and high loss of pesticides. Dermanyssus gallinae is a notorious pest in egg-laying hens, whose control primarily depends on acaricide spraying, while its surface hydrophobicity and potential influence on pesticide effectiveness are not clear. In the present study, the contact angle measurements revealed that the surface of D. gallinae was hydrophobic. Analysis using scanning electron microscopy (SEM) and transmission electron microscopy (TEM) showed that the surface microstructures of D. gallinae consist of cuticular folds, with a lipid-rich outermost layer of the cuticle. Based on gas chromatography-mass spectrometry (GC-MS) and gas chromatography (GC), it was found that the major compositions of cuticular lipids were fatty acids and n-alkanes. Modifying the chemical compositions and microstructures of the D. gallinae surface resulted in a reduction in surface hydrophobicity and an increase in the permeation of Rhodamine B through the cuticle. This observation suggested that the chemical compositions and microstructures were pivotal in determining surface hydrophobicity, hindering compound penetration into the cuticle. Finally, it was found improving the wettability of pesticide solution by adding surfactants could overcome the surface hydrophobicity and enhance the efficacy of pesticide against the mites. This study sheds light on the surface hydrophobicity mechanism of D. gallinae and provides a novel strategy to improve the efficacy of acaricides against the mites.
生物表面的疏水性可提供生物自我保护。害虫表面的疏水性是害虫防治的主要障碍，会导致杀虫剂的低利用率和高损失。Dermanyssus gallinae 是一种臭名昭著的蛋鸡害虫，其防治主要依靠喷洒杀螨剂，而其表面疏水性及其对杀虫剂效果的潜在影响尚不清楚。在本研究中，接触角测量显示五倍子表面疏水。利用扫描电子显微镜（SEM）和透射电子显微镜（TEM）进行的分析表明，五倍子的表面微结构由角质层褶皱组成，角质层最外层富含脂质。根据气相色谱-质谱法（GC-MS）和气相色谱法（GC），发现角质层脂质的主要成分是脂肪酸和正构烷烃。改变五倍子表面的化学成分和微结构可降低表面疏水性，增加罗丹明 B 在角质层中的渗透。这一观察结果表明，化学成分和微观结构在决定表面疏水性方面起着关键作用，阻碍了化合物向角质层的渗透。最后，研究发现通过添加表面活性剂来改善杀虫剂溶液的润湿性，可以克服表面疏水性，提高杀虫剂对螨虫的药效。这项研究揭示了五倍子螨的表面疏水性机理，为提高杀螨剂的药效提供了一种新策略。
{"title":"Surface hydrophobicity mechanism of poultry red mite, Dermanyssus gallinae (Acari: Dermanyssidae), gives novel meaning to chemical control","authors":"Bohan Wang, Jiali Meng, Xiaoxiao Qi, Penglong Wang, Qi Liu, Lifang Wang, Weiwei Sun, Baoliang Pan","doi":"10.1016/j.vetpar.2024.110327","DOIUrl":"10.1016/j.vetpar.2024.110327","url":null,"abstract":"<div><div>Surface hydrophobicity of organisms provides biological self-protection. The hydrophobicity of pest surface, acting as a main obstacle for the pest control, can lead to low utilization and high loss of pesticides. <em>Dermanyssus gallinae</em> is a notorious pest in egg-laying hens, whose control primarily depends on acaricide spraying, while its surface hydrophobicity and potential influence on pesticide effectiveness are not clear. In the present study, the contact angle measurements revealed that the surface of <em>D. gallinae</em> was hydrophobic. Analysis using scanning electron microscopy (SEM) and transmission electron microscopy (TEM) showed that the surface microstructures of <em>D. gallinae</em> consist of cuticular folds, with a lipid-rich outermost layer of the cuticle. Based on gas chromatography-mass spectrometry (GC-MS) and gas chromatography (GC), it was found that the major compositions of cuticular lipids were fatty acids and n-alkanes. Modifying the chemical compositions and microstructures of the <em>D. gallinae</em> surface resulted in a reduction in surface hydrophobicity and an increase in the permeation of Rhodamine B through the cuticle. This observation suggested that the chemical compositions and microstructures were pivotal in determining surface hydrophobicity, hindering compound penetration into the cuticle. Finally, it was found improving the wettability of pesticide solution by adding surfactants could overcome the surface hydrophobicity and enhance the efficacy of pesticide against the mites. This study sheds light on the surface hydrophobicity mechanism of <em>D. gallinae</em> and provides a novel strategy to improve the efficacy of acaricides against the mites.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"332 ","pages":"Article 110327"},"PeriodicalIF":2.0,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142422320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-09DOI: 10.1016/j.vetpar.2024.110321
Muhammad Rashid , Muhammad Hassan Hayat , Naveed Zahra , Mian Sayed Khan , Suleman , Muhammad Nadeem , Tauseef ur Rehman , Muhammad Ehsan , Muhammad Irfan Malik , Muhammad Kashif Obaid , Amir Bakhsh , Mohamed Aziz Darghouth , Qiaoyun Ren
Theileria annulata (T. annulata) is intra-erythrocytic protozoan parasite which is more prevalent in tropical and sub-tropical countries. It has a significant economic impact on the productivity of the dairy industry, and buparvaquone is used to treat infected animals in the prevalent regions of the world. Systematically, buparvaquone targets the cyto-b gene to break the electron transport chain (ETC) and Theileria annulata peptidyl-prolyl isomerase 1 (TaPIN1) gene to destabilize transcription factor JUN (c-JUN) to inhibit proliferation of infected cells, which ultimately leads to the death of T. annulata. The reported studies on drug resistance is due to inappropriate drug application, evolutionary characteristics of the cytochrome b (cyto-b) gene and oncogenic signaling pathways gene (TaPIN1) make the parasite resistant against buparvaquone. Hence, this systematic review was designed to find out non-synonymous mutation in genes (cyto-b and TaPIN1) responsible for drug resistance reported from Tunisia, Turkey, Egypt, Sudan, Iran, Pakistan, China and Germany with reference to the T. annulata Ankara strain of cyto-b (accession no. XM_949625.1) and TaPIN1 (accession no. TA18945) wild type genes. Non-synonymous point mutations were found in cyto-b (Q01 at 130–148 and Q02 at 253–262 regions) and TaPIN1 (A53P and A53T) genes. These point mutations are responsible for developing buparvaquone resistance against T. annulata infection. These genes can be used as biomarkers for the identification of drug resistance in any endemic area. To avoid the complication of drug resistance, development of genetically resistant cattle breeds, potent vaccines and anti-theilerial drugs (Trifloxystrobin and anti-cancerous) are currently required to control proliferating economically important T. annulata parasites.
{"title":"Systematic review on buparvaquone resistance associated with non-synonymous mutation in drug binding genes site of Theileria annulate","authors":"Muhammad Rashid , Muhammad Hassan Hayat , Naveed Zahra , Mian Sayed Khan , Suleman , Muhammad Nadeem , Tauseef ur Rehman , Muhammad Ehsan , Muhammad Irfan Malik , Muhammad Kashif Obaid , Amir Bakhsh , Mohamed Aziz Darghouth , Qiaoyun Ren","doi":"10.1016/j.vetpar.2024.110321","DOIUrl":"10.1016/j.vetpar.2024.110321","url":null,"abstract":"<div><div><em>Theileria annulata</em> (<em>T. annulata</em>) is intra-erythrocytic protozoan parasite which is more prevalent in tropical and sub-tropical countries. It has a significant economic impact on the productivity of the dairy industry, and buparvaquone is used to treat infected animals in the prevalent regions of the world. Systematically, buparvaquone targets the cyto-b gene to break the electron transport chain (ETC) and <em>Theileria annulata</em> peptidyl-prolyl isomerase 1 (TaPIN1) gene to destabilize transcription factor JUN (c-JUN) to inhibit proliferation of infected cells, which ultimately leads to the death of <em>T. annulata</em>. The reported studies on drug resistance is due to inappropriate drug application, evolutionary characteristics of the cytochrome b (cyto-b) gene and oncogenic signaling pathways gene (TaPIN1) make the parasite resistant against buparvaquone. Hence, this systematic review was designed to find out non-synonymous mutation in genes (cyto-b and TaPIN1) responsible for drug resistance reported from Tunisia, Turkey, Egypt, Sudan, Iran, Pakistan, China and Germany with reference to the <em>T. annulata</em> Ankara strain of cyto-b (accession no. XM_949625.1) and TaPIN1 (accession no. TA18945) wild type genes. Non-synonymous point mutations were found in cyto-b (Q<sub>01</sub> at 130–148 and Q<sub>02</sub> at 253–262 regions) and TaPIN1 (A53P and A53T) genes. These point mutations are responsible for developing buparvaquone resistance against <em>T. annulata</em> infection. These genes can be used as biomarkers for the identification of drug resistance in any endemic area. To avoid the complication of drug resistance, development of genetically resistant cattle breeds, potent vaccines and anti-theilerial drugs (Trifloxystrobin and anti-cancerous) are currently required to control proliferating economically important <em>T. annulata</em> parasites.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"332 ","pages":"Article 110321"},"PeriodicalIF":2.0,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142441543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-05DOI: 10.1016/j.vetpar.2024.110326
Andrew C. Kotze , Angela P. Ruffell , Nicholas M. Rolls
Control of the sheep blowfly relies largely on the use of insecticides applied prophylactically in advance of expected fly activity. However, the blowfly has shown an ability to develop resistance to some of these insecticides. Recent reports of the co-occurrence of resistance to both dicyclanil and imidacloprid in in vitro bioassays with field-collected fly strains has raised the possibility that the two resistances may represent cross-resistance linked by a common mechanism. We investigated this by imposing insecticide selection pressure on larvae of two insecticide-resistant field strains over a number of generations using either dicyclanil or imidacloprid and then measuring changes in sensitivity to both the selecting chemical and the alternate chemical. Larvae selected over six generations with each chemical showed significant increases in resistance to the selecting chemical: resistance ratios at the IC50 5.5 - 8.1-fold higher for dicyclanil, and 3.1 - 3.8-fold for imidacloprid. The larvae also showed significant increases in levels of resistance towards the alternate chemical: resistance ratios 2.6 - 3.1-fold higher towards dicyclanil following selection with imidacloprid, and 2.2 - 3.2-fold higher towards imidacloprid following selection with dicyclanil. The increases in resistance to both chemicals after exposure to either suggests a common mechanism of resistance, at least in our laboratory-selected populations. Assays with the cytochrome P450 inhibitor aminobenzotriazole showed that this synergist was able to remove the increased resistance to both compounds in strains selected with either compound, suggesting that cytochrome P450 is responsible for the resistance observed to both chemicals. Our results confirm that cross-resistance exists between dicyclanil and imidacloprid in the sheep blowfly, and hence the two compounds should be considered as related entities in insecticide rotation strategies for flystrike control.
{"title":"Cross-resistance patterns in field strains of the sheep blowfly following laboratory-based selection pressure with dicyclanil or imidacloprid","authors":"Andrew C. Kotze , Angela P. Ruffell , Nicholas M. Rolls","doi":"10.1016/j.vetpar.2024.110326","DOIUrl":"10.1016/j.vetpar.2024.110326","url":null,"abstract":"<div><div>Control of the sheep blowfly relies largely on the use of insecticides applied prophylactically in advance of expected fly activity. However, the blowfly has shown an ability to develop resistance to some of these insecticides. Recent reports of the co-occurrence of resistance to both dicyclanil and imidacloprid in <em>in vitro</em> bioassays with field-collected fly strains has raised the possibility that the two resistances may represent cross-resistance linked by a common mechanism. We investigated this by imposing insecticide selection pressure on larvae of two insecticide-resistant field strains over a number of generations using either dicyclanil or imidacloprid and then measuring changes in sensitivity to both the selecting chemical and the alternate chemical. Larvae selected over six generations with each chemical showed significant increases in resistance to the selecting chemical: resistance ratios at the IC<sub>50</sub> 5.5 - 8.1-fold higher for dicyclanil, and 3.1 - 3.8-fold for imidacloprid. The larvae also showed significant increases in levels of resistance towards the alternate chemical: resistance ratios 2.6 - 3.1-fold higher towards dicyclanil following selection with imidacloprid, and 2.2 - 3.2-fold higher towards imidacloprid following selection with dicyclanil. The increases in resistance to both chemicals after exposure to either suggests a common mechanism of resistance, at least in our laboratory-selected populations. Assays with the cytochrome P450 inhibitor aminobenzotriazole showed that this synergist was able to remove the increased resistance to both compounds in strains selected with either compound, suggesting that cytochrome P450 is responsible for the resistance observed to both chemicals. Our results confirm that cross-resistance exists between dicyclanil and imidacloprid in the sheep blowfly, and hence the two compounds should be considered as related entities in insecticide rotation strategies for flystrike control.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"332 ","pages":"Article 110326"},"PeriodicalIF":2.0,"publicationDate":"2024-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142406960","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Blastocystis is one of the most common intestinal parasites observed in human and non-human hosts. Recent meta-analyses have indicated a potential role for pets such as dogs and cats as reservoir hosts of Blastocystis, but the data underpinning this hypothesis are of mixed quality. Reviewing data for 45,894 samples tested for Blastocystis by DNA-based methods and 11,908 subtype observations, a model was developed for calculating indices that could be used for evaluating individual species as natural hosts of Blastocystis, based on weighted products of positivity rates and subtype distributions. Data from cats and dogs were analysed, using other well-sampled hosts (pig, cattle, sheep, goat, and human) as references. Data from cats and dogs meeting the inclusion criteria were entered into the model. The overall positivity rates for pigs, cattle, sheep, goats, humans, dogs, and cats were 40 %, 40 %, 35 %, 28 %, 25 %, 6 %, and 5 %, respectively, with statistically significant lower positivity rates in cats and dogs (p < 0.0001). Indices indicating Blastocystis specificity to host ranged between 0.16 (humans) and 0.49 (cattle) for the reference hosts, whereas indices for cats and dogs were only 0.01 and 0.02, respectively. Finally, indices for ST specificity to host were higher for reference hosts (range, 0.66–0.93) than for cats (0.62) and dogs (0.56). Taken together, the analyses indicate that cats and dogs are not natural or reservoir hosts of Blastocystis and that the sporadic subtype pattern observed in these hosts might indicate exposure to Blastocystis through contaminated water/feed, including Blastocystis colonizing prey animals.
{"title":"Cats and dogs as hosts of Blastocystis – What is the evidence?","authors":"Supaluk Popruk , Khuanchai Koompapong , Aongart Mahittikorn , Lee O.’Brien Andersen , Christen Rune Stensvold","doi":"10.1016/j.vetpar.2024.110325","DOIUrl":"10.1016/j.vetpar.2024.110325","url":null,"abstract":"<div><div><em>Blastocystis</em> is one of the most common intestinal parasites observed in human and non-human hosts. Recent meta-analyses have indicated a potential role for pets such as dogs and cats as reservoir hosts of <em>Blastocystis</em>, but the data underpinning this hypothesis are of mixed quality. Reviewing data for 45,894 samples tested for <em>Blastocystis</em> by DNA-based methods and 11,908 subtype observations, a model was developed for calculating indices that could be used for evaluating individual species as natural hosts of <em>Blastocystis</em>, based on weighted products of positivity rates and subtype distributions. Data from cats and dogs were analysed, using other well-sampled hosts (pig, cattle, sheep, goat, and human) as references. Data from cats and dogs meeting the inclusion criteria were entered into the model. The overall positivity rates for pigs, cattle, sheep, goats, humans, dogs, and cats were 40 %, 40 %, 35 %, 28 %, 25 %, 6 %, and 5 %, respectively, with statistically significant lower positivity rates in cats and dogs (<em>p</em> < 0.0001). Indices indicating <em>Blastocystis</em> specificity to host ranged between 0.16 (humans) and 0.49 (cattle) for the reference hosts, whereas indices for cats and dogs were only 0.01 and 0.02, respectively. Finally, indices for ST specificity to host were higher for reference hosts (range, 0.66–0.93) than for cats (0.62) and dogs (0.56). Taken together, the analyses indicate that cats and dogs are not natural or reservoir hosts of <em>Blastocystis</em> and that the sporadic subtype pattern observed in these hosts might indicate exposure to <em>Blastocystis</em> through contaminated water/feed, including <em>Blastocystis</em> colonizing prey animals.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"332 ","pages":"Article 110325"},"PeriodicalIF":2.0,"publicationDate":"2024-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142393795","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-04DOI: 10.1016/j.vetpar.2024.110323
José Roberto Pereira , Fernando Garcia Nicodemos , Fernanda Calvo Duarte , Jose Eduardo Marcondes de Almeida , Márcia Cristina Mendes
In the southeastern region of Brazil, ticks of the species Rhipicephalus (B.) microplus are constantly present on cattle throughout the year. This is due to climatic conditions that favor the biology of these ticks and resistance to the acaricides in use that has developed. The aim of this study was to evaluate the efficacy of the fungus Purpureocillium lilacinum (IBCB 130) in powdered form, applied via a drone, from the bioFUNGUS spray dispenser (NCB Sistemas Embarcados Ltda), onto pasture for controlling these ticks. This experiment was conducted at Aptaregional, Regional Research and Development Unit of Pindamonhangaba, São Paulo, Brazil. To evaluate the efficacy of treatment, two groups were formed (treated and control), each with ten calves of the Girolando breed, which were kept in two separate paddocks in which the main grass species was Brachiaria decumbens. The treated paddock received seven monthly applications (November–May) of the fungus Purpureocillium lilacinum (5 g de P. lilacinum in 200 g of wheat flour). The control paddock did not receive any treatment. Tick counts performed on the animals every two weeks showed that, over the entire period, the treatment had a mean efficacy of 48.59 %. The months of December and January presented the highest efficacy rates, of 63.50 % and 83.87 %, respectively.
{"title":"Efficacy of the fungus Purpureocillium lilacinum applied via drone onto pasture for controlling the tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)","authors":"José Roberto Pereira , Fernando Garcia Nicodemos , Fernanda Calvo Duarte , Jose Eduardo Marcondes de Almeida , Márcia Cristina Mendes","doi":"10.1016/j.vetpar.2024.110323","DOIUrl":"10.1016/j.vetpar.2024.110323","url":null,"abstract":"<div><div>In the southeastern region of Brazil, ticks of the species <em>Rhipicephalus</em> (<em>B</em>.) <em>microplus</em> are constantly present on cattle throughout the year. This is due to climatic conditions that favor the biology of these ticks and resistance to the acaricides in use that has developed. The aim of this study was to evaluate the efficacy of the fungus <em>Purpureocillium lilacinum</em> (IBCB 130) in powdered form, applied via a drone, from the bioFUNGUS spray dispenser (NCB Sistemas Embarcados Ltda), onto pasture for controlling these ticks. This experiment was conducted at Aptaregional, Regional Research and Development Unit of Pindamonhangaba, São Paulo, Brazil. To evaluate the efficacy of treatment, two groups were formed (treated and control), each with ten calves of the Girolando breed, which were kept in two separate paddocks in which the main grass species was <em>Brachiaria decumbens</em>. The treated paddock received seven monthly applications (November–May) of the fungus <em>Purpureocillium lilacinum</em> (5 g de <em>P. lilacinum</em> in 200 g of wheat flour). The control paddock did not receive any treatment. Tick counts performed on the animals every two weeks showed that, over the entire period, the treatment had a mean efficacy of 48.59 %. The months of December and January presented the highest efficacy rates, of 63.50 % and 83.87 %, respectively.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"332 ","pages":"Article 110323"},"PeriodicalIF":2.0,"publicationDate":"2024-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142509206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-03DOI: 10.1016/j.vetpar.2024.110324
Alexandre Alves de Sousa Nascimento , Isabella Góes Mantini da Cunha , Marcos Horácio Pereira , Mauricio Roberto Viana Sant’Anna , Alexandre Barbosa Reis , Nelder Figueiredo Gontijo
Dogs are important reservoir hosts for Leishmania infantum, the causative agent of visceral leishmaniasis. The complement system, as part of the innate immune defense, is responsible for initiating the fight against pathogens that may invade an organism. A failure of the complement to combat L. infantum may explain, at least in part, why a mammal species is more or less susceptible to visceral leishmaniasis. The objective of this study was to compare the effectiveness of human and dog complement systems against L. infantum parasites. The results showed that dog serum was less effective than human serum at killing promastigote and amastigote-like forms. We also compared the efficiency of human and canine sera in classic and alternative hemolytic assays, as well as the serum efficiency of non-infected and Leishmania-infected dogs. Serum from dogs was less hemolytic than human serum in both pathways tested, but the efficiency of serum from infected dogs was higher than that of non-infected dogs. When testing C3b deposition assays on parasite surfaces, serum from infected dogs was more effective against amastigote-like forms than serum from non-infected dogs. However, both types of serum proved equally effective on promastigotes, while serum from infected dogs was more effective on amastigote-like forms. Considering the efficiency of the complement system, our results indicate that dogs are more susceptible to visceral leishmaniasis than humans are.
{"title":"Dog complement system is less effective against Leishmania infantum than human complement","authors":"Alexandre Alves de Sousa Nascimento , Isabella Góes Mantini da Cunha , Marcos Horácio Pereira , Mauricio Roberto Viana Sant’Anna , Alexandre Barbosa Reis , Nelder Figueiredo Gontijo","doi":"10.1016/j.vetpar.2024.110324","DOIUrl":"10.1016/j.vetpar.2024.110324","url":null,"abstract":"<div><div>Dogs are important reservoir hosts for <em>Leishmania infantum</em>, the causative agent of visceral leishmaniasis. The complement system, as part of the innate immune defense, is responsible for initiating the fight against pathogens that may invade an organism. A failure of the complement to combat <em>L. infantum</em> may explain, at least in part, why a mammal species is more or less susceptible to visceral leishmaniasis. The objective of this study was to compare the effectiveness of human and dog complement systems against <em>L. infantum</em> parasites. The results showed that dog serum was less effective than human serum at killing promastigote and amastigote-like forms. We also compared the efficiency of human and canine sera in classic and alternative hemolytic assays, as well as the serum efficiency of non-infected and <em>Leishmania</em>-infected dogs. Serum from dogs was less hemolytic than human serum in both pathways tested, but the efficiency of serum from infected dogs was higher than that of non-infected dogs. When testing C3b deposition assays on parasite surfaces, serum from infected dogs was more effective against amastigote-like forms than serum from non-infected dogs. However, both types of serum proved equally effective on promastigotes, while serum from infected dogs was more effective on amastigote-like forms. Considering the efficiency of the complement system, our results indicate that dogs are more susceptible to visceral leishmaniasis than humans are.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"332 ","pages":"Article 110324"},"PeriodicalIF":2.0,"publicationDate":"2024-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142381759","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-02DOI: 10.1016/j.vetpar.2024.110322
Yanning Li , Tongxuan Zhang , Xuanrui Liu , Tianxu Pan , Junyi Li , Wentao Yang , Xin Cao , Yanlong Jiang , Jianzhong Wang , Yan Zeng , Chunwei Shi , Haibin Huang , Chunfeng Wang , Nan Wang , Guilian Yang
The Apicomplexa parasitic phylum rhoptry neck protein 2 (RON2) plays a key role in the process of invading host cells. Eimeria tenella, an intracellular protozoan shares a similar conserved invasion pattern. However, whether E. tenella RON2 participates in the process of invading the host intestinal epithelium is poorly understood. In this study, the sequence of EtRON2 was analyzed and expressed. The expression of the truncated extracellular N-terminal fragment of EtRON2 (403–700 aa, designated EtRON2403–700) with a molecular mass of 38.3 kDa. EtRON2 in the sporozoite protein was detected at 151.4 kDa by rabbit anti-rEtRON2403–700 antibody. Immunofluorescence results showed that EtRON2 was mainly localized to the nucleus and apex of the E. tenella sporozoite. qPCR results showed that the highest expression level of EtRON2 was detected in sporulated oocysts compared with other developmental stages of E. tenella. In vitro invasion inhibition assays showed that the capacity of sporozoites to invade DF-1 cells was significantly inhibited after pretreatment with the rabbit anti-rEtRON2403–700 antibody. Silencing the EtRON2 gene by RNA interference (RNAi) significantly inhibited EtRON2 expression and significantly reduced the invasion of DF-1 cells by sporozoites. In vivo experiments revealed a significant decrease parasite burden and oocyst outputs in chicks after infection with EtRON2 gene-silenced sporozoites by cloacal inoculation. Recombinant EtRON2403–700 (rEtRON2403–700) immunizes chicks effectively against E. tenella infection by inducing humoral immunity and upregulating IFN-γ and CD8+ T lymphocytes. Furthermore, chicks exhibited increased relative weight gain rates, lower cecum lesion scores, and reduced oocyst outputs during the E. tenella challenge. H&E staining showed that the cecum tissue of chicks immunized with rEtRON2403–700 showed relatively mild histopathological changes. In conclusion, the results of this study demonstrated that EtRON2 plays a key role in E. tenella invasion of the host intestinal epithelium and provides a potential target for vaccines against E. tenella infection.
{"title":"Eimeria tenella rhoptry neck protein 2 plays a key role in the process of invading the host intestinal epithelium","authors":"Yanning Li , Tongxuan Zhang , Xuanrui Liu , Tianxu Pan , Junyi Li , Wentao Yang , Xin Cao , Yanlong Jiang , Jianzhong Wang , Yan Zeng , Chunwei Shi , Haibin Huang , Chunfeng Wang , Nan Wang , Guilian Yang","doi":"10.1016/j.vetpar.2024.110322","DOIUrl":"10.1016/j.vetpar.2024.110322","url":null,"abstract":"<div><div>The Apicomplexa parasitic phylum rhoptry neck protein 2 (RON2) plays a key role in the process of invading host cells. <em>Eimeria tenella</em>, an intracellular protozoan shares a similar conserved invasion pattern. However, whether <em>E. tenella</em> RON2 participates in the process of invading the host intestinal epithelium is poorly understood. In this study, the sequence of <em>Et</em>RON2 was analyzed and expressed. The expression of the truncated extracellular N-terminal fragment of <em>Et</em>RON2 (403–700 aa, designated <em>Et</em>RON2<sub>403–700</sub>) with a molecular mass of 38.3 kDa. <em>Et</em>RON2 in the sporozoite protein was detected at 151.4 kDa by rabbit anti-r<em>Et</em>RON2<sub>403–700</sub> antibody. Immunofluorescence results showed that <em>Et</em>RON2 was mainly localized to the nucleus and apex of the <em>E. tenella</em> sporozoite. qPCR results showed that the highest expression level of <em>Et</em>RON2 was detected in sporulated oocysts compared with other developmental stages of <em>E. tenella</em>. In vitro invasion inhibition assays showed that the capacity of sporozoites to invade DF-1 cells was significantly inhibited after pretreatment with the rabbit anti-r<em>Et</em>RON2<sub>403–700</sub> antibody. Silencing the <em>Et</em>RON2 gene by RNA interference (RNAi) significantly inhibited <em>Et</em>RON2 expression and significantly reduced the invasion of DF-1 cells by sporozoites. In vivo experiments revealed a significant decrease parasite burden and oocyst outputs in chicks after infection with <em>Et</em>RON2 gene-silenced sporozoites by cloacal inoculation. Recombinant <em>Et</em>RON2<sub>403–700</sub> (r<em>Et</em>RON2<sub>403–700</sub>) immunizes chicks effectively against <em>E. tenella</em> infection by inducing humoral immunity and upregulating IFN-γ and CD8<sup>+</sup> T lymphocytes. Furthermore, chicks exhibited increased relative weight gain rates, lower cecum lesion scores, and reduced oocyst outputs during the <em>E. tenella</em> challenge. H&E staining showed that the cecum tissue of chicks immunized with r<em>Et</em>RON2<sub>403–700</sub> showed relatively mild histopathological changes. In conclusion, the results of this study demonstrated that <em>Et</em>RON2 plays a key role in <em>E. tenella</em> invasion of the host intestinal epithelium and provides a potential target for vaccines against <em>E. tenella</em> infection.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"332 ","pages":"Article 110322"},"PeriodicalIF":2.0,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142376117","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-01DOI: 10.1016/j.vetpar.2024.110320
Muhammed Ahmed Selcuk , Burcak Aslan Celik , Figen Celik , Ozgur Yasar Celik , Kerem Ercan , Muhammet Uslug , Afra Sena Tekin , Sami Simsek
Cystic echinococcosis (CE), caused by Echinococcus granulosus sensu lato (s.l.), represents one of the most significant zoonotic diseases globally, affecting both humans and animals. The objective of this study was to ascertain the prevalence of E. granulosus sensu lato in sheep and goats in a pilot region with a one-year slaughterhouse follow-up period and to determine the genetic differences and haplotypes among sheep, goat, and dog isolates. To this end, the prevalence of CE cysts was determined by monitoring the slaughter of sheep and goats at least three days a week at a slaughterhouse in the Siirt province of Türkiye during 2023. Additionally, faecal samples were collected from stray dogs and analysed using both flotation and molecular techniques. The presence of CE cysts was identified in 569 (11.12 %) of the 5119 sheep and 66 (2.31 %) of the 2860 goats after slaughtering. The highest positivity was observed in November (20.39 %), while the lowest was recorded in July (5.62 %). Of the sheep that detected positive, 25 (4.39 %) were less than one year old, while 544 (95.61 %) were older than one year. Of the infected sheep, 26 (4.57 %) were male and 543 (95.43 %) were female. 204 (35.85 %) sheep exhibited fluid-filled CE cysts, 338 (59.40 %) displayed calcification, and 27 (4.75 %) demonstrated the presence of newly developed cysts. The highest positivity was observed in December (5.83 %), while the lowest was recorded in May (0.62 %) in goats. Of the positive goats, two (3 %) were less than one year old, while the remaining 64 (97 %) were older than one year. Of the goats infected with CE cysts, 10 (15.15 %) were male and 56 (84.85 %) were female. Of the cysts, 56.1 % were fluid-filled, 42.4 % were calcified and 1.5 % were newly developed. Following DNA sequence analysis of CE cyst isolates obtained from the slaughterhouse, all 61 sheep sequences were identified as E. granulosus s.s. (G1/G3). Of the 13 goat isolates, seven were identified as E. granulosus s.s. (G1/G3), while the remaining six were classified as E. canadensis (G6/G7). The centrifugal flotation method was employed to detect the presence of Isospora spp. oocysts in eight dogs, Toxocara canis and hookworm eggs in three dogs each, and Dipyllidium caninum eggs in one dog. A total of 54 dog faeces were examined. No Taeniid eggs were observed in any of the dogs. Following PCR analysis of the mt-CO1 gene region in the dog faecal samples, four samples were positive for a 875 bp band. Only one of these bands was suitable for sequence analysis, which confirmed it as E. granulosus s.s. (G1/G3).
{"title":"A pilot study on the epidemiology, diagnosis and characterization of Echinococcus granulosus sensu lato in sheep, goats and dogs in Siirt province of Türkiye revealed remarkable adaptation of Echinococcus canadensis (G6/G7) in goats","authors":"Muhammed Ahmed Selcuk , Burcak Aslan Celik , Figen Celik , Ozgur Yasar Celik , Kerem Ercan , Muhammet Uslug , Afra Sena Tekin , Sami Simsek","doi":"10.1016/j.vetpar.2024.110320","DOIUrl":"10.1016/j.vetpar.2024.110320","url":null,"abstract":"<div><div>Cystic echinococcosis (CE), caused by <em>Echinococcus granulosus</em> sensu lato (s.l.), represents one of the most significant zoonotic diseases globally, affecting both humans and animals. The objective of this study was to ascertain the prevalence of <em>E. granulosus</em> sensu lato in sheep and goats in a pilot region with a one-year slaughterhouse follow-up period and to determine the genetic differences and haplotypes among sheep, goat, and dog isolates. To this end, the prevalence of CE cysts was determined by monitoring the slaughter of sheep and goats at least three days a week at a slaughterhouse in the Siirt province of Türkiye during 2023. Additionally, faecal samples were collected from stray dogs and analysed using both flotation and molecular techniques. The presence of CE cysts was identified in 569 (11.12 %) of the 5119 sheep and 66 (2.31 %) of the 2860 goats after slaughtering. The highest positivity was observed in November (20.39 %), while the lowest was recorded in July (5.62 %). Of the sheep that detected positive, 25 (4.39 %) were less than one year old, while 544 (95.61 %) were older than one year. Of the infected sheep, 26 (4.57 %) were male and 543 (95.43 %) were female. 204 (35.85 %) sheep exhibited fluid-filled CE cysts, 338 (59.40 %) displayed calcification, and 27 (4.75 %) demonstrated the presence of newly developed cysts. The highest positivity was observed in December (5.83 %), while the lowest was recorded in May (0.62 %) in goats. Of the positive goats, two (3 %) were less than one year old, while the remaining 64 (97 %) were older than one year. Of the goats infected with CE cysts, 10 (15.15 %) were male and 56 (84.85 %) were female. Of the cysts, 56.1 % were fluid-filled, 42.4 % were calcified and 1.5 % were newly developed. Following DNA sequence analysis of CE cyst isolates obtained from the slaughterhouse, all 61 sheep sequences were identified as <em>E. granulosus</em> s.s. (G1/G3). Of the 13 goat isolates, seven were identified as <em>E. granulosus</em> s.s. (G1/G3), while the remaining six were classified as <em>E. canadensis</em> (G6/G7). The centrifugal flotation method was employed to detect the presence of <em>Isospora</em> spp. oocysts in eight dogs, <em>Toxocara canis</em> and hookworm eggs in three dogs each, and <em>Dipyllidium caninum</em> eggs in one dog. A total of 54 dog faeces were examined. No Taeniid eggs were observed in any of the dogs. Following PCR analysis of the mt-CO1 gene region in the dog faecal samples, four samples were positive for a 875 bp band. Only one of these bands was suitable for sequence analysis, which confirmed it as <em>E. granulosus</em> s.s. (G1/G3).</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"332 ","pages":"Article 110320"},"PeriodicalIF":2.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142393794","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-29DOI: 10.1016/j.vetpar.2024.110319
Maria Alfonsa Cavalera , Annamaria Uva , Floriana Gernone , Oana Gusatoaia , Rossella Donghia , Andrea Zatelli
This prospective, randomized, controlled, therapeutic study aimed to evaluate the efficacy of a product containing nucleotides and lactoferrin in maintaining or improving the clinical picture and laboratory findings of canine leishmaniosis (CanL). The safety and tolerance of this combination were also assessed. Forty Leishmania infantum-seropositive dogs, not requiring leishmanicidal and/or leishmaniostatic treatment, were enrolled in the study and randomized into treatment (TG) and placebo (CG) groups. Products A (containing nucleotides and lactoferrin) and B (placebo) were blindly administered to TG and CG, respectively, as palatable tablets at a rate of 1 tablet per 10 kg of weight once every 24 h for 6 months. Following inclusion (T0), dogs were followed up after 3 (T90) and 6 (T180) months. At each time point, for all animals enrolled physical examination and laboratory tests (complete blood count, biochemical panel including C-reactive protein [CRP] and ferritin, and serum protein electrophoresis) were performed. The immunofluorescence antibody test to detect antibodies for L. infantum (T0, T180), Ehrlichia canis (T0, T90, and T180), and Anaplasma phagocytophilum (T0, T90, and T180) was executed. A CanL-dedicated clinical score, using a validated scale from 0 (i.e., absence of clinical signs) to 19, was assigned. Four dogs (n=2 in TG, n=2 in CG) did not complete the study. No statistically significant differences in CanL clinical score were observed between CG and TG at T0, T90 and T180. Both TG and CG showed significant variations in anti-L. infantum antibody titres (p=0.0001 and p=0.004, respectively). In TG, antibody titres decreased in 77.8 %, increased in 5.5 %, and remained stable in 16.7 % of dogs, while in CG, decreased in 27.8 %, increased in 50 %, and remained stable in 22.2 % of dogs. During the study, CRP and ferritin remained stable in TG and significantly increased in CG. At T180, 9 out of 18 dogs (50 %) enrolled in the CG, and 1 out of 18 (5.6 %) enrolled in the TG, developed an active form of leishmaniosis. No side effects were reported in any patient included. In conclusion, a 6-month oral administration of a supplement containing nucleotides and lactoferrin was effective in maintaining a stable clinical score, improving antibody titres and potentially reducing the progression from non-active to active forms in L. infatum seropositive dogs. Furthermore, the product was well-tolerated, easy to administer, and free of side effects.
{"title":"Efficacy of a combination of nucleotides and lactoferrin in maintaining stable or improving the clinical picture and laboratory findings of leishmaniotic dogs: A randomized controlled study","authors":"Maria Alfonsa Cavalera , Annamaria Uva , Floriana Gernone , Oana Gusatoaia , Rossella Donghia , Andrea Zatelli","doi":"10.1016/j.vetpar.2024.110319","DOIUrl":"10.1016/j.vetpar.2024.110319","url":null,"abstract":"<div><div>This prospective, randomized, controlled, therapeutic study aimed to evaluate the efficacy of a product containing nucleotides and lactoferrin in maintaining or improving the clinical picture and laboratory findings of canine leishmaniosis (CanL). The safety and tolerance of this combination were also assessed. Forty <em>Leishmania infantum</em>-seropositive dogs, not requiring leishmanicidal and/or leishmaniostatic treatment, were enrolled in the study and randomized into treatment (TG) and placebo (CG) groups. Products A (containing nucleotides and lactoferrin) and B (placebo) were blindly administered to TG and CG, respectively, as palatable tablets at a rate of 1 tablet per 10 kg of weight once every 24 h for 6 months. Following inclusion (T0), dogs were followed up after 3 (T90) and 6 (T180) months. At each time point, for all animals enrolled physical examination and laboratory tests (complete blood count, biochemical panel including C-reactive protein [CRP] and ferritin, and serum protein electrophoresis) were performed. The immunofluorescence antibody test to detect antibodies for <em>L. infantum</em> (T0, T180), <em>Ehrlichia canis</em> (T0, T90, and T180), and <em>Anaplasma phagocytophilum</em> (T0, T90, and T180) was executed. A CanL-dedicated clinical score, using a validated scale from 0 (i.e., absence of clinical signs) to 19, was assigned. Four dogs (n=2 in TG, n=2 in CG) did not complete the study. No statistically significant differences in CanL clinical score were observed between CG and TG at T0, T90 and T180. Both TG and CG showed significant variations in anti-<em>L. infantum</em> antibody titres (p=0.0001 and p=0.004, respectively). In TG, antibody titres decreased in 77.8 %, increased in 5.5 %, and remained stable in 16.7 % of dogs, while in CG, decreased in 27.8 %, increased in 50 %, and remained stable in 22.2 % of dogs. During the study, CRP and ferritin remained stable in TG and significantly increased in CG. At T180, 9 out of 18 dogs (50 %) enrolled in the CG, and 1 out of 18 (5.6 %) enrolled in the TG, developed an active form of leishmaniosis. No side effects were reported in any patient included. In conclusion, a 6-month oral administration of a supplement containing nucleotides and lactoferrin was effective in maintaining a stable clinical score, improving antibody titres and potentially reducing the progression from non-active to active forms in <em>L. infatum</em> seropositive dogs. Furthermore, the product was well-tolerated, easy to administer, and free of side effects.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"332 ","pages":"Article 110319"},"PeriodicalIF":2.0,"publicationDate":"2024-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142376116","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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