Mastitis in cows is one of the most important diseases that give rise to economic losses in dairy farms. Increasing antimicrobial resistance in Staphylococcus aureus, one of the most common causes of mastitis, is a significant health problem. Due to the problems encountered in treating infections caused by resistant strains, developing alternative treatment methods, such as Nanomaterial systems and natural agents, are important. The essential oil of Melaleuca alternifolia is used as an antibacterial and the primary active component is terpinen-4-ol. This study aimed to investigate the antibacterial and anti-biofilm activity of terpinen-4-ol and terpinen-4-ol loaded polydopamine (T-PDA) nanoparticles against S. aureus isolates, which were resistant to at least one group of antibiotics isolated from milk samples of subclinical mastitis cows. The S. aureus strains were identified by biochemical tests and verified with the API Staph kit. The antibiotic susceptibility of the isolates was determined by the disc diffusion method. The broth microdilution method determined the antimicrobial activities of the terpinen-4-ol and T-PDA nanoparticles, and anti-biofilm activities were assessed using the modified crystal violet method. All of the isolates were resistant to benzylpenicillin and susceptible to trimethoprim/sulfamethoxazole. Multi-antibiotic resistance was detected in the 11 S. aureus isolates used in this study. For the terpinen-4-ol and T-PDA nanoparticles, MIC values were determined in the range of 0.125-0.5% (µL/mL) and 0.125-0.25% (µL/mL), respectively. None of the isolates formed biofilms. As a result, it was found that the antibacterial efficacy of the T- PDA nanoparticles was higher against nine of the S. aureus isolates than against the terpinen-4-ol.
奶牛乳腺炎是造成奶牛场经济损失的最重要疾病之一。金黄色葡萄球菌是最常见的乳腺炎病因之一,其抗菌药耐药性的增加是一个重大的健康问题。由于在治疗耐药菌株引起的感染时遇到的问题,开发替代治疗方法(如纳米材料系统和天然制剂)非常重要。互叶白千层的精油被用作抗菌剂,其主要活性成分是萜品烯-4-醇。本研究旨在调查萜品烯-4-醇和萜品烯-4-醇负载聚多巴胺(T-PDA)纳米粒子对金葡菌分离株的抗菌和抗生物膜活性。这些金黄色葡萄球菌菌株通过生化测试进行鉴定,并用 API Staph 试剂盒进行验证。分离菌株对抗生素的敏感性是通过盘扩散法测定的。肉汤微稀释法测定了萜品烯-4-醇和 T-PDA 纳米粒子的抗菌活性,改良水晶紫法评估了抗生物膜活性。所有分离菌株均对苄青霉素耐药,对三甲双胍/磺胺甲噁唑易感。在本研究中使用的 11 个金黄色葡萄球菌分离物中检测到了多种抗生素耐药性。萜品烯-4-醇和 T-PDA 纳米粒子的 MIC 值分别为 0.125-0.5%(微升/毫升)和 0.125-0.25%(微升/毫升)。没有一种分离物形成生物膜。结果发现,T-PDA 纳米粒子对九种金黄色葡萄球菌分离物的抗菌效力高于对 4-羟基松油的抗菌效力。
{"title":"Determination of antibacterial and anti-biofilm activities of Terpinen-4-ol loaded polydopamine nanoparticles against Staphylococcus aureus isolates from cows with subclinical mastitis.","authors":"Nilgün Ünal, Merve Eylul Kiymaci, Merve Savluk, Hakan Erdogan, Esra Seker","doi":"10.1007/s11259-024-10514-w","DOIUrl":"10.1007/s11259-024-10514-w","url":null,"abstract":"<p><p>Mastitis in cows is one of the most important diseases that give rise to economic losses in dairy farms. Increasing antimicrobial resistance in Staphylococcus aureus, one of the most common causes of mastitis, is a significant health problem. Due to the problems encountered in treating infections caused by resistant strains, developing alternative treatment methods, such as Nanomaterial systems and natural agents, are important. The essential oil of Melaleuca alternifolia is used as an antibacterial and the primary active component is terpinen-4-ol. This study aimed to investigate the antibacterial and anti-biofilm activity of terpinen-4-ol and terpinen-4-ol loaded polydopamine (T-PDA) nanoparticles against S. aureus isolates, which were resistant to at least one group of antibiotics isolated from milk samples of subclinical mastitis cows. The S. aureus strains were identified by biochemical tests and verified with the API Staph kit. The antibiotic susceptibility of the isolates was determined by the disc diffusion method. The broth microdilution method determined the antimicrobial activities of the terpinen-4-ol and T-PDA nanoparticles, and anti-biofilm activities were assessed using the modified crystal violet method. All of the isolates were resistant to benzylpenicillin and susceptible to trimethoprim/sulfamethoxazole. Multi-antibiotic resistance was detected in the 11 S. aureus isolates used in this study. For the terpinen-4-ol and T-PDA nanoparticles, MIC values were determined in the range of 0.125-0.5% (µL/mL) and 0.125-0.25% (µL/mL), respectively. None of the isolates formed biofilms. As a result, it was found that the antibacterial efficacy of the T- PDA nanoparticles was higher against nine of the S. aureus isolates than against the terpinen-4-ol.</p>","PeriodicalId":23690,"journal":{"name":"Veterinary Research Communications","volume":" ","pages":"3655-3668"},"PeriodicalIF":1.8,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142081812","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2024-12-01Epub Date: 2024-09-21DOI: 10.1007/s11259-024-10548-0
Nayne Vieira da Silva, Roberto Bernardino Júnior, Quintiliano Siqueira Schroden Nomelini, Gabriella Faria Pereira, Diego José Zanzarini Delfiol, Geison Morel Nogueira
The passive stay apparatus works by blocking flexion of the knee and tarsus joints in the horse, preventing muscle fatigue. During ambulation, if this mechanism fails to release, the animal will present upward fixation of the patella, which in severe cases, can only be treated by medial patellar desmotomy (MPD). The objective of this study was to investigate the possible electromyographic and behavioral changes, after the impairment of the passive stay apparatus, in horses who have undergone MPD surgery. Five horses presenting dorsal fixation of the patella underwent electromyographic measurements at preoperatively and immediately postoperatively, 24 h, seven, 14 and 21 days after surgery. The samples were collected on a computerized surface electromyograph designed in accordance with the standards of international society of electrophysiology and kinesiology (ISEK). To capture the signals, electrodes positioned on the tensor fasciae lata, vastus lateralis, biceps femoris and semitendinosus muscles were used, bilaterally. These muscles were chosen due to their participation in the passive stay apparatus. The electromyographic signals were collected and processed using a software calibrated to collect data with a sampling frequency of 1000 Hz per channel. The collections took place over 60 min periods, and the results were obtained from the root means square (RMS) analysis. It was noted that the preoperative average of rest in tripedal support was 62%. On the other hand, change in weight bearing member and rest in three limbs in the postoperative period did not occur. This inability to maintain tripedal support in the postoperative period prevents the horse from resting while standing. Furthermore, there was a gradual increase in the need for post-surgical muscle recruitment, resulting in long periods of decubitus due to muscle fatigue, which is less than ideal for such large animals. Therefore, it was concluded that the animals submitted to medial patellar desmotomy, in this study, presented an inability to maintain tripedal support, in addition to a greater need for recruitment of muscle fibers, evidenced by the gradual increase in the electromyographic tracing and total RMS value of the muscles evaluated.
{"title":"Electromyographic and behavioral analysis of horses submitted to medial patellar desmotomy.","authors":"Nayne Vieira da Silva, Roberto Bernardino Júnior, Quintiliano Siqueira Schroden Nomelini, Gabriella Faria Pereira, Diego José Zanzarini Delfiol, Geison Morel Nogueira","doi":"10.1007/s11259-024-10548-0","DOIUrl":"10.1007/s11259-024-10548-0","url":null,"abstract":"<p><p>The passive stay apparatus works by blocking flexion of the knee and tarsus joints in the horse, preventing muscle fatigue. During ambulation, if this mechanism fails to release, the animal will present upward fixation of the patella, which in severe cases, can only be treated by medial patellar desmotomy (MPD). The objective of this study was to investigate the possible electromyographic and behavioral changes, after the impairment of the passive stay apparatus, in horses who have undergone MPD surgery. Five horses presenting dorsal fixation of the patella underwent electromyographic measurements at preoperatively and immediately postoperatively, 24 h, seven, 14 and 21 days after surgery. The samples were collected on a computerized surface electromyograph designed in accordance with the standards of international society of electrophysiology and kinesiology (ISEK). To capture the signals, electrodes positioned on the tensor fasciae lata, vastus lateralis, biceps femoris and semitendinosus muscles were used, bilaterally. These muscles were chosen due to their participation in the passive stay apparatus. The electromyographic signals were collected and processed using a software calibrated to collect data with a sampling frequency of 1000 Hz per channel. The collections took place over 60 min periods, and the results were obtained from the root means square (RMS) analysis. It was noted that the preoperative average of rest in tripedal support was 62%. On the other hand, change in weight bearing member and rest in three limbs in the postoperative period did not occur. This inability to maintain tripedal support in the postoperative period prevents the horse from resting while standing. Furthermore, there was a gradual increase in the need for post-surgical muscle recruitment, resulting in long periods of decubitus due to muscle fatigue, which is less than ideal for such large animals. Therefore, it was concluded that the animals submitted to medial patellar desmotomy, in this study, presented an inability to maintain tripedal support, in addition to a greater need for recruitment of muscle fibers, evidenced by the gradual increase in the electromyographic tracing and total RMS value of the muscles evaluated.</p>","PeriodicalId":23690,"journal":{"name":"Veterinary Research Communications","volume":" ","pages":"4153-4158"},"PeriodicalIF":1.8,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142296713","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2024-12-01Epub Date: 2024-09-20DOI: 10.1007/s11259-024-10544-4
Pablo Oyarzún-Ruiz, Mauricio A Navarro, Sergio Donoso-Erch, Sebastián Muñoz-Leal
Syngamid worms (Nematoda: Syngamidae) parasitizing birds of prey are considered cosmopolitan, but the efforts to understand their biology and systematics are restricted to the Holarctic region. However, in the Neotropical region there is only one recent record with no data about its molecular characterization or its significance to the health of its hosts. Thus, this study aimed to identify through an integrative approach the Syngamid worms parasitizing a native owl, and to describe its pathological consequences. A total of 14 barn owls (Tyto alba) were dissected between 2015 and 2021, from which one bird was found to be parasitized by a high parasitic burden (i.e. 185 worms). Considering light microscopy and SEM, these nematodes were morphologically identified as Cyathostoma americana, and then supported through molecular analyses of nuclear loci ITS and LSU. The pathological changes were described as severe airsacculitis and pneumonia, which probably were the cause of death of the bird. This study represents the first effort to characterize the parasitism by this nematode in a nocturnal bird of prey from the Neotropics, with a lethal outcome. Additional studies are required from other geographical areas and Neotropical countries to better understand the dynamics of this parasite and their hosts, considering both diurnal and nocturnal birds of prey.
寄生于鸟类的鞘翅目蠕虫(线虫纲:Syngamidae)被认为是世界性的,但了解其生物学和系统学的工作仅限于全北极地区。然而,在新热带地区只有一个最新的记录,没有关于其分子特征或其对宿主健康的重要性的数据。因此,本研究旨在通过综合方法确定寄生在本地猫头鹰身上的 Syngamid 蠕虫,并描述其病理后果。研究人员在2015年至2021年期间共解剖了14只仓鸮(Tyto alba),发现其中一只鸟寄生了大量寄生虫(即185条蠕虫)。通过光学显微镜和扫描电子显微镜,这些线虫在形态学上被鉴定为美洲盘尾丝虫,并通过核位点 ITS 和 LSU 的分子分析得到支持。病理变化被描述为严重的气囊炎和肺炎,这可能是导致该鸟死亡的原因。这项研究首次描述了这种线虫在新热带地区夜行性猎鸟体内寄生并导致死亡的情况。为了更好地了解这种寄生虫及其宿主的动态,还需要对其他地区和新热带国家的昼行性和夜行性猎禽进行更多的研究。
{"title":"Integrative description and histopathological lesions of Cyathostoma americana (Nematoda, Syngamidae) parasitizing a barn owl (Tyto alba) in Chile.","authors":"Pablo Oyarzún-Ruiz, Mauricio A Navarro, Sergio Donoso-Erch, Sebastián Muñoz-Leal","doi":"10.1007/s11259-024-10544-4","DOIUrl":"10.1007/s11259-024-10544-4","url":null,"abstract":"<p><p>Syngamid worms (Nematoda: Syngamidae) parasitizing birds of prey are considered cosmopolitan, but the efforts to understand their biology and systematics are restricted to the Holarctic region. However, in the Neotropical region there is only one recent record with no data about its molecular characterization or its significance to the health of its hosts. Thus, this study aimed to identify through an integrative approach the Syngamid worms parasitizing a native owl, and to describe its pathological consequences. A total of 14 barn owls (Tyto alba) were dissected between 2015 and 2021, from which one bird was found to be parasitized by a high parasitic burden (i.e. 185 worms). Considering light microscopy and SEM, these nematodes were morphologically identified as Cyathostoma americana, and then supported through molecular analyses of nuclear loci ITS and LSU. The pathological changes were described as severe airsacculitis and pneumonia, which probably were the cause of death of the bird. This study represents the first effort to characterize the parasitism by this nematode in a nocturnal bird of prey from the Neotropics, with a lethal outcome. Additional studies are required from other geographical areas and Neotropical countries to better understand the dynamics of this parasite and their hosts, considering both diurnal and nocturnal birds of prey.</p>","PeriodicalId":23690,"journal":{"name":"Veterinary Research Communications","volume":" ","pages":"4141-4147"},"PeriodicalIF":1.8,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142296714","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2024-12-01Epub Date: 2024-09-24DOI: 10.1007/s11259-024-10545-3
Eliz Oliveira Franco, Francisco Chagas Bezerra Dos Santos, Rair de Sousa Verde, Ana Cláudia Calchi, Victória Valente Califre de Mello, Daniel Braga Lee, Clara Morato Dias, Rosangela Zacarias Machado, Adolorata Aparecida Bianco Carvalho, André Luiz Rodrigues Roque, Marcos Rogério André
Despite the great diversity of bats (64 species) in the State of Acre, northwestern Brazil, there are no studies on occurrence and diversity of Bartonella spp. in bats in this region. The present study investigated the occurrence and molecular identity of Bartonella spp. in spleen samples (n = 271) from bats of 30 different species from this region, within the Amazon biome. Twenty-one out of 208 (10.1%) samples positive in the PCR for the mammalian gapdh endogenous genes were positive in the qPCR for Bartonella spp. based on the nuoG gene. The two gltA Bartonella genotypes detected grouped with those previously identified in bats from other locations, expanding the diversity of genotypes associated with bats. This study provided the first molecular evidence of the presence of Bartonella spp. in bats in the state of Acre and in bats of the species Lophostoma silvicolum, Vampyressa thyone, Tonatia saurophila and Phyllostomus elongatus.
{"title":"Bartonella spp. in bats from the Brazilian Amazon Forest.","authors":"Eliz Oliveira Franco, Francisco Chagas Bezerra Dos Santos, Rair de Sousa Verde, Ana Cláudia Calchi, Victória Valente Califre de Mello, Daniel Braga Lee, Clara Morato Dias, Rosangela Zacarias Machado, Adolorata Aparecida Bianco Carvalho, André Luiz Rodrigues Roque, Marcos Rogério André","doi":"10.1007/s11259-024-10545-3","DOIUrl":"10.1007/s11259-024-10545-3","url":null,"abstract":"<p><p>Despite the great diversity of bats (64 species) in the State of Acre, northwestern Brazil, there are no studies on occurrence and diversity of Bartonella spp. in bats in this region. The present study investigated the occurrence and molecular identity of Bartonella spp. in spleen samples (n = 271) from bats of 30 different species from this region, within the Amazon biome. Twenty-one out of 208 (10.1%) samples positive in the PCR for the mammalian gapdh endogenous genes were positive in the qPCR for Bartonella spp. based on the nuoG gene. The two gltA Bartonella genotypes detected grouped with those previously identified in bats from other locations, expanding the diversity of genotypes associated with bats. This study provided the first molecular evidence of the presence of Bartonella spp. in bats in the state of Acre and in bats of the species Lophostoma silvicolum, Vampyressa thyone, Tonatia saurophila and Phyllostomus elongatus.</p>","PeriodicalId":23690,"journal":{"name":"Veterinary Research Communications","volume":" ","pages":"4159-4168"},"PeriodicalIF":1.8,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142308639","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2024-12-01Epub Date: 2024-09-28DOI: 10.1007/s11259-024-10560-4
Luiza Aymée, Ana Luiza Dos Santos Baptista Borges, Guilherme Nunes de Souza, Walter Lilenbaum
Bovine Genital Leptospirosis (BGL) is a chronic reproductive syndrome characterized by genital infection by Leptospira spp. An accurate diagnosis of BGL is crucial to implementing proper control measures in field conditions. This study aimed to evaluate the reliability of serology by Microscopic Agglutination Test (MAT) for diagnosing leptospirosis in subfertile cows with genital infection. Of three herds, 93 non-pregnant cows with reproductive failures were submitted to the blood sampling (serology by MAT) and genital samples (lipL32-PCR). A total of 62/93 (66.6%) cows presented seroreactive to cutoff 100, while 45/93 (48.4%) cows were positive to cutoff 200, mainly against the Sejroe serogroup. In PCR analysis, 55/93 (59.1%) were positive. MAT results were compared with PCR (considered the standard), and test parameters and Cohen's kappa (ƙ) were calculated for the cut-offs 100 and 200. A ROC curve was performed for each cut-off of titers 100 to 1,600. The sensitivity and specificity of MAT100 were calculated at 66.6% and 33.3%, while for MAT200 the sensitivity was estimated as 35% and specificity as 54.5%. The accuracy of MAT was poor, being 54.8% in MAT100 and 42% in MAT200. Furthermore, the area under the curve of ROC analysis was low for all titers, and the correlation was poor for MAT100 and MAT200 (ƙ < 0). The results demonstrated that MAT is a limited technique to diagnose bovine genital carriers individually, and if only MAT is applied, genital carriers may pass undetected, impairing the control programs.
{"title":"Is microscopic agglutination test a reliable method for diagnosing the bovine genital leptospirosis syndrome?","authors":"Luiza Aymée, Ana Luiza Dos Santos Baptista Borges, Guilherme Nunes de Souza, Walter Lilenbaum","doi":"10.1007/s11259-024-10560-4","DOIUrl":"10.1007/s11259-024-10560-4","url":null,"abstract":"<p><p>Bovine Genital Leptospirosis (BGL) is a chronic reproductive syndrome characterized by genital infection by Leptospira spp. An accurate diagnosis of BGL is crucial to implementing proper control measures in field conditions. This study aimed to evaluate the reliability of serology by Microscopic Agglutination Test (MAT) for diagnosing leptospirosis in subfertile cows with genital infection. Of three herds, 93 non-pregnant cows with reproductive failures were submitted to the blood sampling (serology by MAT) and genital samples (lipL32-PCR). A total of 62/93 (66.6%) cows presented seroreactive to cutoff 100, while 45/93 (48.4%) cows were positive to cutoff 200, mainly against the Sejroe serogroup. In PCR analysis, 55/93 (59.1%) were positive. MAT results were compared with PCR (considered the standard), and test parameters and Cohen's kappa (ƙ) were calculated for the cut-offs 100 and 200. A ROC curve was performed for each cut-off of titers 100 to 1,600. The sensitivity and specificity of MAT100 were calculated at 66.6% and 33.3%, while for MAT200 the sensitivity was estimated as 35% and specificity as 54.5%. The accuracy of MAT was poor, being 54.8% in MAT100 and 42% in MAT200. Furthermore, the area under the curve of ROC analysis was low for all titers, and the correlation was poor for MAT100 and MAT200 (ƙ < 0). The results demonstrated that MAT is a limited technique to diagnose bovine genital carriers individually, and if only MAT is applied, genital carriers may pass undetected, impairing the control programs.</p>","PeriodicalId":23690,"journal":{"name":"Veterinary Research Communications","volume":" ","pages":"4185-4189"},"PeriodicalIF":1.8,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142355061","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2024-12-01Epub Date: 2024-10-08DOI: 10.1007/s11259-024-10541-7
Gamil S G Zeedan, Abeer M Abdalhamed, Ahmad M Allam, Sobhy Abdel-Shafy
Lumpy skin disease (LSD) is a viral disease that affects cattle and buffaloes in Egypt, causing considerable economic losses in the animal sector. This study aimed to investigate the recent outbreak of LSDV in cattle and buffaloes and evaluate the potential role of the hard tick Rhipicephalus annulatus in their transmission through isolation and molecular characterization by multiplex PCR (mPCR) and real-time quantitative PCR (rt-qPCR) assays. A total of 50 skin biopsies (cattle n = 30, buffaloes n = 20), 110 nasal swabs (cattle n = 76, buffaloes n = 44), and 129 blood samples (cattle n = 84, buffaloes n = 45) were collected. In addition, 145 hard ticks of different stages were collected from cattle and buffaloes of different breeds and ages in different governorates in Egypt from November 2021 to June 2022. Multiplex PCR and real-time quantitative PCR (rt-qPCR) assays based on SYBR Green and targets (P32, VP32, G protein, and viral fusion protein) were used. We identified positive results in 17 out of 30 cattle skin biopsies (56.6%), 1 out of 7 buffalo skin scabs (14.3%), and 5 out of 45 buffalo blood samples (11.11%) using mPCR and RT-qPCR methods. We successfully isolated LSDV from hard ticks and cattle infested with ticks and exhibited characteristic signs of LSD on the chorioallantois membrane (CAM) of specific pathogen-free embryonated chicken eggs (SPF-ECE). The isolates were confirmed by multiplex PCR and RT-qPCR. The cyclic threshold (Ct) with correlation-slandered curve values of rt-qPCR ranging from 10.2 to 36.5 showed the amount of LSDV-DNA in different samples. The study's findings demonstrated the widespread circulation of LSDV in both cattle and buffaloes in Egypt and provided strong evidence that hard ticks R. annulatus play a role in the transmission of LSDV in susceptible animals.
{"title":"Molecular detection of lumpy skin disease virus in naturally infected cattle and buffaloes: unveiling the role of tick vectors in disease spread.","authors":"Gamil S G Zeedan, Abeer M Abdalhamed, Ahmad M Allam, Sobhy Abdel-Shafy","doi":"10.1007/s11259-024-10541-7","DOIUrl":"10.1007/s11259-024-10541-7","url":null,"abstract":"<p><p>Lumpy skin disease (LSD) is a viral disease that affects cattle and buffaloes in Egypt, causing considerable economic losses in the animal sector. This study aimed to investigate the recent outbreak of LSDV in cattle and buffaloes and evaluate the potential role of the hard tick Rhipicephalus annulatus in their transmission through isolation and molecular characterization by multiplex PCR (mPCR) and real-time quantitative PCR (rt-qPCR) assays. A total of 50 skin biopsies (cattle n = 30, buffaloes n = 20), 110 nasal swabs (cattle n = 76, buffaloes n = 44), and 129 blood samples (cattle n = 84, buffaloes n = 45) were collected. In addition, 145 hard ticks of different stages were collected from cattle and buffaloes of different breeds and ages in different governorates in Egypt from November 2021 to June 2022. Multiplex PCR and real-time quantitative PCR (rt-qPCR) assays based on SYBR Green and targets (P32, VP32, G protein, and viral fusion protein) were used. We identified positive results in 17 out of 30 cattle skin biopsies (56.6%), 1 out of 7 buffalo skin scabs (14.3%), and 5 out of 45 buffalo blood samples (11.11%) using mPCR and RT-qPCR methods. We successfully isolated LSDV from hard ticks and cattle infested with ticks and exhibited characteristic signs of LSD on the chorioallantois membrane (CAM) of specific pathogen-free embryonated chicken eggs (SPF-ECE). The isolates were confirmed by multiplex PCR and RT-qPCR. The cyclic threshold (Ct) with correlation-slandered curve values of rt-qPCR ranging from 10.2 to 36.5 showed the amount of LSDV-DNA in different samples. The study's findings demonstrated the widespread circulation of LSDV in both cattle and buffaloes in Egypt and provided strong evidence that hard ticks R. annulatus play a role in the transmission of LSDV in susceptible animals.</p>","PeriodicalId":23690,"journal":{"name":"Veterinary Research Communications","volume":" ","pages":"3921-3939"},"PeriodicalIF":1.8,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11538203/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142393777","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-12-01Epub Date: 2024-10-23DOI: 10.1007/s11259-024-10563-1
Eman Fayez, Haney Samir, Fady Sayed Youssef, Ali Salama, Mohamed Ai ElSayed
Environmental heat stress (HS) impairs reproductive efficiency in farm animals. This study investigated, for the first time, how the melatonin and melatonin nanoparticles treatment affected the testicular hemodynamics, testicular volume, echotexture [Pixel intensity (PIX) and integrated density (IND)], scrotal circumference, serum concentration of testosterone (T), estradiol (E2), nitric oxide (NO), and total antioxidant capacity (TAC) in prepubertal Ossimi ram lambs in hot climatic conditions. The lambs undergoing examination had a temperature humidity index (THI) of 87.05 ± 1.70, indicating severe HS condition. Fifteen prepubertal Ossimi ram lambs were exposed to a single s.c injection of either nano melatonin (nano melatonin group; 20 mg/ram; n = 5) or melatonin suspended in two ml of corn oil (melatonin group; 40 mg/ram; n = 5) or two ml of corn oil (control group; n = 5). Blood collection and ultrasonographic assessment of the testes and supratesticular arteries (STAs) were conducted immediately before treatment (W0) and once weekly for six successive weeks after nano melatonin and melatonin injection (W1-W6). Results revealed decreases (P < 0.05) in the Doppler indices (resistive index; RI and pulsatility index; PI) of the testicular arteries at most time points of the study in the nano melatonin and melatonin groups. PIX of testicular parenchyma was significantly increased (P ˂ 0.05) in the treated groups compared to the control one. IND of testicular parenchyma increased significantly in the nano melatonin group compared to the melatonin and control groups. Testicular volume and scrotal circumference significantly increased (P < 0.05) in nano melatonin and melatonin groups compared to the control one. T concentration did not significantly (P > 0.05) change in the treated groups compared to the control group. E2, NO, and TAC concentrations increased (P < 0.05) in the treated groups compared to the control one. In conclusion, this study extrapolated that administrations of melatonin or nano melatonin can ameliorate the effects of environmental HS in prepubertal Ossimi ram lambs with a more protective effect and lower dose of nano melatonin.
{"title":"Administration of melatonin nanoparticles improves testicular blood flow, echotexture of testicular parenchyma, scrotal circumference, and levels of estradiol and nitric oxide in prepubertal ossimi rams under summer heat stress.","authors":"Eman Fayez, Haney Samir, Fady Sayed Youssef, Ali Salama, Mohamed Ai ElSayed","doi":"10.1007/s11259-024-10563-1","DOIUrl":"10.1007/s11259-024-10563-1","url":null,"abstract":"<p><p>Environmental heat stress (HS) impairs reproductive efficiency in farm animals. This study investigated, for the first time, how the melatonin and melatonin nanoparticles treatment affected the testicular hemodynamics, testicular volume, echotexture [Pixel intensity (PIX) and integrated density (IND)], scrotal circumference, serum concentration of testosterone (T), estradiol (E<sub>2</sub>), nitric oxide (NO), and total antioxidant capacity (TAC) in prepubertal Ossimi ram lambs in hot climatic conditions. The lambs undergoing examination had a temperature humidity index (THI) of 87.05 ± 1.70, indicating severe HS condition. Fifteen prepubertal Ossimi ram lambs were exposed to a single s.c injection of either nano melatonin (nano melatonin group; 20 mg/ram; n <sub>=</sub> 5) or melatonin suspended in two ml of corn oil (melatonin group; 40 mg/ram; n <sub>=</sub> 5) or two ml of corn oil (control group; n <sub>=</sub> 5). Blood collection and ultrasonographic assessment of the testes and supratesticular arteries (STAs) were conducted immediately before treatment (W0) and once weekly for six successive weeks after nano melatonin and melatonin injection (W1-W6). Results revealed decreases (P < 0.05) in the Doppler indices (resistive index; RI and pulsatility index; PI) of the testicular arteries at most time points of the study in the nano melatonin and melatonin groups. PIX of testicular parenchyma was significantly increased (P ˂ 0.05) in the treated groups compared to the control one. IND of testicular parenchyma increased significantly in the nano melatonin group compared to the melatonin and control groups. Testicular volume and scrotal circumference significantly increased (P < 0.05) in nano melatonin and melatonin groups compared to the control one. T concentration did not significantly (P > 0.05) change in the treated groups compared to the control group. E<sub>2</sub>, NO, and TAC concentrations increased (P < 0.05) in the treated groups compared to the control one. In conclusion, this study extrapolated that administrations of melatonin or nano melatonin can ameliorate the effects of environmental HS in prepubertal Ossimi ram lambs with a more protective effect and lower dose of nano melatonin.</p>","PeriodicalId":23690,"journal":{"name":"Veterinary Research Communications","volume":" ","pages":"3953-3965"},"PeriodicalIF":1.8,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11538174/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142509150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-12-01Epub Date: 2024-09-03DOI: 10.1007/s11259-024-10528-4
Alexandra Bergen, Savannah Roemhild, Domenico Santoro
Skin infections are common complications in both humans and animals. Because of the increased incidence of multi-drug resistant (MDR) skin infections, essential oils have been suggested as potential alternatives to the classic antimicrobials. The goal of this study was to evaluate the minimum inhibitory and bactericidal/fungicidal concentrations (MIC and MBC/MFC) of commercially available products containing essential oils, zinc gluconate, or 4% chlorhexidine. Microbroth dilution technique was performed on clinical isolates of MDR Staphylococcus pseudintermedius (MDR-SP; n = 10), Pseudomonas aeruginosa (PA; n = 10), and Malassezia pachydermatis (MP; n = 10). For MDR-SP, essential oil-containing products showed median MICs of 1:240 and 1:320. The chlorhexidine shampoo had a MIC of 1:128,000 (0.312 µg/mL), whereas zinc gluconate products had median MICs of 1:320 and 1:160. Three essential oil-containing shampoos (MBC 1:40), the zinc gluconate (MBC 1:40), and the chlorhexidine (MBC 1:64,000 [0.625 µg/mL]) reached an MBC. For PA, essential oil-containing products showed median MICs of 1:30 and 1:80. The zinc-gluconate products had a median MIC of 1:160, whereas the chlorhexidine shampoo had a median MIC of 1:4,000 (10 µg/mL). Only the zinc-gluconate products (MBC 1:80) and the chlorhexidine shampoo (MBC 1:2,000 [20 µg/mL]) reached an MBC. For MP, essential oil-containing and zinc-gluconate products showed lower median MICs (1:4,800 and 7,200) for shampoos compared with other formulations (1:160 and 1:320), whereas the chlorhexidine shampoo had a median MIC of 1:80,000 (0.5 µg/mL). These results suggest that natural topical compounds can be an effective alternative to treat skin infections in companion animals. Further in vivo studies are needed to clinically confirm this study's results.
皮肤感染是人类和动物常见的并发症。由于耐多药(MDR)皮肤感染的发病率越来越高,人们建议用精油作为传统抗菌剂的潜在替代品。本研究的目的是评估含有精油、葡萄糖酸锌或 4% 洗必泰的市售产品的最低抑菌和杀菌/杀真菌浓度(MIC 和 MBC/MFC)。对临床分离出的 MDR 假中间葡萄球菌(MDR-SP;n = 10)、铜绿假单胞菌(PA;n = 10)和马拉色菌(MP;n = 10)进行了微流稀释技术处理。对于 MDR-SP,含精油产品的 MIC 中位数分别为 1:240 和 1:320。洗必泰洗发水的 MIC 值为 1:128,000(0.312 µg/mL),而葡萄糖酸锌产品的中位 MIC 值为 1:320 和 1:160。三种含精油的洗发水(MBC 1:40)、葡萄糖酸锌(MBC 1:40)和洗必泰(MBC 1:64,000 [0.625 µg/mL])达到了 MBC。就 PA 而言,含精油产品的 MIC 中位数分别为 1:30 和 1:80。葡萄糖酸锌产品的 MIC 中位数为 1:160,而洗必泰洗发水的 MIC 中位数为 1:4,000(10 µg/mL)。只有葡萄糖酸锌产品(MBC 1:80)和洗必泰洗发水(MBC 1:2,000 [20 µg/mL])达到了 MBC。就 MP 而言,与其他配方(1:160 和 1:320)相比,含精油和葡萄糖酸锌产品的洗发水 MIC 中位数较低(1:4800 和 7,200),而洗必泰洗发水的 MIC 中位数为 1:80000(0.5 µg/mL)。这些结果表明,天然外用化合物是治疗伴侣动物皮肤感染的有效替代品。要在临床上证实这项研究的结果,还需要进一步的体内研究。
{"title":"Minimum inhibitory and bactericidal/fungicidal concentration of commercially available products containing essential oils, zinc gluconate, or 4% chlorhexidine for Malassezia pachydermatis, Pseudomonas aeruginosa, and multi-drug resistant Staphylococcus pseudintermedius canine clinical isolates.","authors":"Alexandra Bergen, Savannah Roemhild, Domenico Santoro","doi":"10.1007/s11259-024-10528-4","DOIUrl":"10.1007/s11259-024-10528-4","url":null,"abstract":"<p><p>Skin infections are common complications in both humans and animals. Because of the increased incidence of multi-drug resistant (MDR) skin infections, essential oils have been suggested as potential alternatives to the classic antimicrobials. The goal of this study was to evaluate the minimum inhibitory and bactericidal/fungicidal concentrations (MIC and MBC/MFC) of commercially available products containing essential oils, zinc gluconate, or 4% chlorhexidine. Microbroth dilution technique was performed on clinical isolates of MDR Staphylococcus pseudintermedius (MDR-SP; n = 10), Pseudomonas aeruginosa (PA; n = 10), and Malassezia pachydermatis (MP; n = 10). For MDR-SP, essential oil-containing products showed median MICs of 1:240 and 1:320. The chlorhexidine shampoo had a MIC of 1:128,000 (0.312 µg/mL), whereas zinc gluconate products had median MICs of 1:320 and 1:160. Three essential oil-containing shampoos (MBC 1:40), the zinc gluconate (MBC 1:40), and the chlorhexidine (MBC 1:64,000 [0.625 µg/mL]) reached an MBC. For PA, essential oil-containing products showed median MICs of 1:30 and 1:80. The zinc-gluconate products had a median MIC of 1:160, whereas the chlorhexidine shampoo had a median MIC of 1:4,000 (10 µg/mL). Only the zinc-gluconate products (MBC 1:80) and the chlorhexidine shampoo (MBC 1:2,000 [20 µg/mL]) reached an MBC. For MP, essential oil-containing and zinc-gluconate products showed lower median MICs (1:4,800 and 7,200) for shampoos compared with other formulations (1:160 and 1:320), whereas the chlorhexidine shampoo had a median MIC of 1:80,000 (0.5 µg/mL). These results suggest that natural topical compounds can be an effective alternative to treat skin infections in companion animals. Further in vivo studies are needed to clinically confirm this study's results.</p>","PeriodicalId":23690,"journal":{"name":"Veterinary Research Communications","volume":" ","pages":"3699-3709"},"PeriodicalIF":1.8,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142120699","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2024-12-01Epub Date: 2024-09-05DOI: 10.1007/s11259-024-10512-y
Osama Ewais, Heba Abdel-Tawab, Huda El-Fayoumi, Shawky M Aboelhadid, Saleh Al-Quraishy, Piotr Falkowski, Abdel-Azeem S Abdel-Baki
The excessive use of conventional medications to treat coccidiosis has led to concerns regarding drug residues in tissues and the emergence of multidrug resistance. Essential oils with anti-inflammatory and antioxidant activities may also have anticoccidial effects. The present study investigated the efficacy of D-limonene and its nanoemulsion form against Eimeria tenella in chickens. An in vitro study was conducted to evaluate the sporulation inhibitory effects of D-limonene on Eimeria tenella oocysts. Five D-limonene concentrations (0.625, 1.25, 2.5, 5, and 10% v/v) were tested alongside positive (10% formalin) and negative (2.5% potassium dichromate) controls. Each ELISA plate well was inoculated with 1200 unsporulated oocysts and incubated at 30 °C for 24, 48, and 72 h. Subsequently, samples were microscopically examined to assess sporulation inhibition and calculate the percentage of sporulated oocysts. For the in vivo study, 125 eight-day-old broiler chicks were divided into five groups of 25 birds each. The control negative group remained uninfected and untreated. The control positive group was challenged with 5 × 104 sporulated Eimeria tenella oocysts. The diclazuril group received 0.2 mg/kg diclazuril in their diet two days prior to, and until 10 days post infection. The D-limonene (DL) and D-limonene nanoemulsion (DLN) groups were challenged with 5 × 104 sporulated E. tenella oocysts at 18 days of age and administered 150 mg/L of their respective treatments in drinking water from day eight until the end of the experiment. Results from the in vitro study demonstrated that D-limonene suppressed oocyst sporulation by 50.83% at its highest concentration of 10%. In the in vivo study, both DL and DLN treated groups exhibited a significant reduction in oocyst output per gram of feces (OPG), along with increased body weight and decreased parasite stages in the cecal tissue. Furthermore, these treatments were associated with elevated levels of antioxidant enzymes such as glutathione peroxidase (GPX), catalase (CAT), and superoxide dismutase (SOD), accompanied by a decrease in malondialdehyde (MDA) and nitric oxide (NO) levels. Particularly, DLN treatment remarkably increased the number of goblet cells. In conclusion, D-limonene and its nanoemulsion represent promising alternatives for managing coccidiosis in poultry. They not only effectively control parasites but also promote intestinal health and boost antioxidant defenses.
{"title":"Antioxidant properties of D-limonene and its nanoemulsion form enhance its anticoccidial efficiency in experimentally infected broilers with Eimeria tenella: an in vitro and in vivo study.","authors":"Osama Ewais, Heba Abdel-Tawab, Huda El-Fayoumi, Shawky M Aboelhadid, Saleh Al-Quraishy, Piotr Falkowski, Abdel-Azeem S Abdel-Baki","doi":"10.1007/s11259-024-10512-y","DOIUrl":"10.1007/s11259-024-10512-y","url":null,"abstract":"<p><p>The excessive use of conventional medications to treat coccidiosis has led to concerns regarding drug residues in tissues and the emergence of multidrug resistance. Essential oils with anti-inflammatory and antioxidant activities may also have anticoccidial effects. The present study investigated the efficacy of D-limonene and its nanoemulsion form against Eimeria tenella in chickens. An in vitro study was conducted to evaluate the sporulation inhibitory effects of D-limonene on Eimeria tenella oocysts. Five D-limonene concentrations (0.625, 1.25, 2.5, 5, and 10% v/v) were tested alongside positive (10% formalin) and negative (2.5% potassium dichromate) controls. Each ELISA plate well was inoculated with 1200 unsporulated oocysts and incubated at 30 °C for 24, 48, and 72 h. Subsequently, samples were microscopically examined to assess sporulation inhibition and calculate the percentage of sporulated oocysts. For the in vivo study, 125 eight-day-old broiler chicks were divided into five groups of 25 birds each. The control negative group remained uninfected and untreated. The control positive group was challenged with 5 × 10<sup>4</sup> sporulated Eimeria tenella oocysts. The diclazuril group received 0.2 mg/kg diclazuril in their diet two days prior to, and until 10 days post infection. The D-limonene (DL) and D-limonene nanoemulsion (DLN) groups were challenged with 5 × 10<sup>4</sup> sporulated E. tenella oocysts at 18 days of age and administered 150 mg/L of their respective treatments in drinking water from day eight until the end of the experiment. Results from the in vitro study demonstrated that D-limonene suppressed oocyst sporulation by 50.83% at its highest concentration of 10%. In the in vivo study, both DL and DLN treated groups exhibited a significant reduction in oocyst output per gram of feces (OPG), along with increased body weight and decreased parasite stages in the cecal tissue. Furthermore, these treatments were associated with elevated levels of antioxidant enzymes such as glutathione peroxidase (GPX), catalase (CAT), and superoxide dismutase (SOD), accompanied by a decrease in malondialdehyde (MDA) and nitric oxide (NO) levels. Particularly, DLN treatment remarkably increased the number of goblet cells. In conclusion, D-limonene and its nanoemulsion represent promising alternatives for managing coccidiosis in poultry. They not only effectively control parasites but also promote intestinal health and boost antioxidant defenses.</p>","PeriodicalId":23690,"journal":{"name":"Veterinary Research Communications","volume":" ","pages":"3711-3725"},"PeriodicalIF":1.8,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142133923","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 : 2024-12-01Epub Date: 2024-09-21DOI: 10.1007/s11259-024-10550-6
Seda Ekici, Ogun Bozkaya, Sedat Sevin, Babur Erdem, Okan Can Arslan, Ozge Özgenç Cinar, Esra Bozkaya, Husamettin Ekici
In this study, silver nanoparticles (AgNPs) coated with bioactive molecules were synthesized via Vitex agnus-castus L. (VAC) seed extract (VAC/AgNPs). The synthesized VAC/AgNPs were characterized by surface plasmon resonance (SPR) ultraviolet-visible region spectroscopy (UV-Vis). The hydrodynamic size and surface charge analysis of the particles were measured with a Zeta sizer. The results of UV-Vis and Zeta analysis revealed that AgNPs were synthesized, the size distribution was nanoscale, and the solution was stable. The effects of the synthesized VAC/AgNPs and aqueous extract of VAC seeds on honeybees were investigated by means of lifespan test and histopathological analysis. It was determined that both VAC seed extract and VAC/AgNPs were non-toxic to honeybees at certain doses, positively affected their life span and contributed to their longevity in the life span test. Furthermore, no adverse effects were detected in terms of intestinal health in histopathological examinations. Therefore, VAC/AgNPs are considered to be a promising bioactive agent for honeybees.
{"title":"Investigation of the biological activity and toxicity of bioactive silver nanoparticles synthesized via Vitex agnus-castus seed extract on honey bees.","authors":"Seda Ekici, Ogun Bozkaya, Sedat Sevin, Babur Erdem, Okan Can Arslan, Ozge Özgenç Cinar, Esra Bozkaya, Husamettin Ekici","doi":"10.1007/s11259-024-10550-6","DOIUrl":"10.1007/s11259-024-10550-6","url":null,"abstract":"<p><p>In this study, silver nanoparticles (AgNPs) coated with bioactive molecules were synthesized via Vitex agnus-castus L. (VAC) seed extract (VAC/AgNPs). The synthesized VAC/AgNPs were characterized by surface plasmon resonance (SPR) ultraviolet-visible region spectroscopy (UV-Vis). The hydrodynamic size and surface charge analysis of the particles were measured with a Zeta sizer. The results of UV-Vis and Zeta analysis revealed that AgNPs were synthesized, the size distribution was nanoscale, and the solution was stable. The effects of the synthesized VAC/AgNPs and aqueous extract of VAC seeds on honeybees were investigated by means of lifespan test and histopathological analysis. It was determined that both VAC seed extract and VAC/AgNPs were non-toxic to honeybees at certain doses, positively affected their life span and contributed to their longevity in the life span test. Furthermore, no adverse effects were detected in terms of intestinal health in histopathological examinations. Therefore, VAC/AgNPs are considered to be a promising bioactive agent for honeybees.</p>","PeriodicalId":23690,"journal":{"name":"Veterinary Research Communications","volume":" ","pages":"3813-3821"},"PeriodicalIF":1.8,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142296715","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}