Rickettsial infection in animals and Brazilian spotted fever endemicity

We compared the rickettsial infection status of Amblyomma cajennense ticks, humans, dogs, and horses in both Brazilian spotted fever (BSF)-endemic and -nonendemic areas in the state of Sao Paulo, Brazil. Most of the horses and few dogs from BSF-endemic areas had serologic titers against Rickettsia rickettsii antigens. In contrast, no dogs or horses from BSF-nonendemic areas had serologic titers against R. rickettsii antigens, although they were continually exposed to A. cajennense ticks. All human serum samples and ticks from both areas were negative by serologic assay and polymerase chain reaction, respectively. Our results indicate that surveys of horse serum are a useful method of BSF surveillance in areas where humans are exposed to A. cajennense ticks. In addition, we successfully performed experimental infection of A. cajennense ticks with R. parkeri.     

Serosurvey of Rickettsia spp. in dogs and humans from an endemic area for Brazilian spotted fever in the State of São Paulo, Brazil

The present study provides a rickettsial serosurvey in 25 dogs and 35 humans in an endemic area for Brazilian spotted fever in the State of S?o Paulo, where the tick Amblyomma aureolatum is the main vector. Testing canine and human sera by indirect immunofluorescence against four Rickettsia antigens (R. rickettsii, R. parkeri, R. felis and R. bellii) showed that 16 (64%) of canine sera and 1 (2.8%) of human sera reacted to at least one of these rickettsial antigens with titers >0r= 64. Seven canine sera and the single reactive human serum showed titers to R. rickettsii at least four times those of any of the other three antigens. The antibody titers in these 7 animals and 1 human were attributed to stimulation by R. rickettsii infection. No positive canine or human serum was attributed to stimulation by R. parkeri, R. felis, or R. bellii. Our serological results showed that dogs are important sentinels for the presence of R. rickettsii in areas where the tick A. aureolatum is the main vector of Brazilian spotted fever.     

Equine piroplasmosis associated with Amblyomma cajennense Ticks, Texas, USA

We report an outbreak of equine piroplasmosis in southern Texas, USA, in 2009. Infection prevalence reached 100% in some areas (292 infected horses). Amblyomma cajennense was the predominant tick and experimentally transmitted Theileria equi to an uninfected horse. We suggest that transmission by this tick species played a role in this outbreak.     

新疆伊犁地区马群中博尔纳病病毒自然感染调查

目的 了解新疆伊犁地区马群中博尔纳病病毒(BDV)的流行状况,分析该病毒的种系来源.方法 采用改进的巢式反转录PCR(nRT-PCR)方法对新疆伊犁地区120匹马的外周血单个核细胞(PBMC)及脑组织中的BDV p24基因片段进行检测,对阳性产物进行基因序列测定和同源性分析.结果 有3匹马外周血和腩组织中同时检测到BDV,阳性率为2.5%(3/120).扩增产物序列与其他国外马源BDV分离株同源性>93%,与标准株He/80同源性达到98%以上.结论 新疆伊犁地区马群中存在BDV的自然感染.该地区BDV流行株与标准株He/80存在高度的同源性.     

立氏立克次体实时荧光定量聚合酶链反应检测方法的建立

目的采用TaqMan-MGB探针建立立氏立克次体实时荧光定量聚合酶链反应（PCR）检测方法.方法依据立氏立克次体外膜蛋白B基因序列设计引物和探针,以克隆的ompB基因片段为DNA模板,在ABI 7900型荧光定量PCR检测仪上建立实时荧光定量PCR检测方法.结果建立的定量标准曲线Ct值与模板拷贝数呈线性关系（R2=0.996）,最低检测浓度为5拷贝/μl;用荧光定量PCR方法检测其他相关立克次体和常见非立克次体病原菌,检出结果均为阴性.用该方法检测立氏立克次体感染的豚鼠血液标本、小鼠脾脏标本及细胞培养标本,检测的结果与立氏立克次体感染相关.结论研究建立的检测立氏立克次体实时荧光定量PCR方法具有高特异性和高敏感性,适用于快速检测各种样本中的立氏立克次体和立氏立克次体感染早期的实验室诊断.     

Postepizootic persistence of Venezuelan equine encephalitis virus, Venezuela

Five years after the apparent end of the major 1995 Venezuelan equine encephalitis (VEE) epizootic/epidemic, focal outbreaks of equine encephalitis occurred in Carabobo and Barinas States of western Venezuela. Virus isolates from horses in each location were nearly identical in sequence to 1995 isolates, which suggests natural persistence of subtype IC VEE virus (VEEV) strains in a genetically stable mode. Serologic evidence indicated that additional outbreaks occurred in Barinas State in 2003. Field studies identified known Culex (Melanoconion) spp. vectors and reservoir hosts of enzootic VEEV but a dearth of typical epidemic vectors. Cattle serosurveys indicated the recent circulation of enzootic VEEV strains, and possibly of epizootic strains. Persistence of VEEV subtype IC strains and infection of horses at the end of the rainy season suggest the possibility of an alternative, cryptic transmission cycle involving survival through the dry season of infected vectors or persistently infected vertebrates.     

Infection dynamics of Theileria equi in carrier horses is associated with management and tick exposure

The tick-borne equine hemoparasite, Theileria equi, is endemic in many parts of the world where prevalence may be high, and most infected horses are apparently healthy but serve as life-long carriers. To determine the factors that affect T. equi dynamics, we followed parasitic loads in apparently healthy horses at four time points during one year. A total of 1094 blood samples were collected from 395 horses, along with ticks and demographic and clinical data. Infection and load of T. equi were tested by PCR and qPCR, and for the spring dataset, infection was also tested serologically by IFAT (n = 268). Theileria equi was molecularly detected in 64.8 % of the horses. The agreement between molecular and serological results was 79.8 % (K > 0.674) and positively correlated with parasitic load. Infection was associated with pale mucus membranes, lower packed cell volume and higher total solids (all P < 0.001), although these changes had only minor clinical importance. While parasitic loads in qPCR-positive samples (n = 561) were generally low (mean = 7.9−10⁻⁴% parasitized erythrocytes), younger horses showed higher loads, possibly suggesting development of immunity. Infection and parasitic load were associated with housing management and tick exposure, illustrating different patterns of exposure. Endemic stability is suggested in pasture farms with constant exposure to ticks, where parasite prevalence was high (96 %) and associated with T. equi 18S rRNA genotype D, low parasitemia and high antibody titers. Endemic instability can be suggested in case were horses are kept in paddocks (prevalence = 49 %) with intermittent exposure to ticks, where infection was associated with high parasitemia when ticks were present. A steady state is suggested in stabled horses (prevalence = 46 %), with no exposure to ticks; where infection was associated with genotype A, low parasitemia and low antibody titers. The ability to identify different risk groups within endemic areas may improve the administration of suitable treatment and control practices in an effort to reduce the risk of clinical disease.     

Experimental Rhodococcus equi and equine infectious anemia virus DNA vaccination in adult and neonatal horses: effect of IL-12, dose, and route

Improving the ability of DNA-based vaccines to induce potent Type1/Th1 responses against intracellular pathogens in large outbred species is essential. Rhodoccocus equi and equine infectious anemia virus (EIAV) are two naturally occurring equine pathogens that also serve as important large animal models of neonatal immunity and lentiviral immune control. Neonates present a unique challenge for immunization due to their diminished immunologic capabilities and apparent Th2 bias. In an effort to augment R. equi- and EIAV-specific Th1 responses induced by DNA vaccination, we hypothesized that a dual promoter plasmid encoding recombinant equine IL-12 (rEqIL-12) would function as a molecular adjuvant. In adult horses, DNA vaccines induced R. equi- and EIAV-specific antibody and lymphoproliferative responses, and EIAV-specific CTL and tetramer-positive CD8+ T lymphocytes. These responses were not enhanced by the rEqIL-12 plasmid. In neonatal foals, DNA immunization induced EIAV-specific antibody and lymphoproliferative responses, but not CTL. The R. equi vapA vaccine was poorly immunogenic in foals even when co-administered with the IL-12 plasmid. It was concluded that DNA immunization was capable of inducing Th1 responses in horses; dose and route were significant variables, but rEqIL-12 was not an effective molecular adjuvant. Additional work is needed to optimize DNA vaccine-induced Th1 responses in horses, especially in neonates.     

首次实验室证实北京平谷地区恙虫病东方体暴发流行

目的了解北京市平谷地区是否存在恙虫病东方体感染。方法采用巢式聚合酶链反应（nPCR）,对平谷地区采集的30份临床标本进行恙虫病东方体热休克蛋白（groEL）基因和56×103蛋白扩增并测序分析。采用间接免疫荧光试验（IFA）对血清标本恙虫病东方体特异抗体检测。结果采用PCR共检测血液标本30份,均阳性（100%）;用IFA检测28份患者血清抗体,25份阳性（89.3%）;长片段PCR扩增,3份标本扩增阳性,阳性结果测序比对所有核苷酸序列相同,与Kawasaki的同源性为96%。结论首次从分子流行病学和血清流行病学的角度证实北京平谷地区存在恙虫病东方体。     

First identification and phylogenetic analysis of equine hepacivirus in Korea

Non-primate hepacivirus (NPHV) corresponds a group of isolates recently characterized in horses and dogs that present similar genomic organization and are closely related to hepatitis C virus. Since canine hapacivirus, NPHV identified in dogs, was first discovered in dogs in the United States, equine hepacivirus (EqHV, NPHV identified in horses) has been identified in horses in several countries. However, no epidemiological studies have investigated EqHV in horses in Korea. In this study, a total of 74 (n = 74) serum samples collected from horses in four regions of Korea were tested for EqHV RNA using nested RT-PCR. Overall, 14 samples were identified as positive (18.9%) and further analyzed according to gender, age, breed, and region. There were high positive rates in males, young horses, and Thoroughbreds; however, these rates differed regionally. Sequencing of the partial NS3 region of 12 samples and the polyprotein encoding regions of two samples positive for EqHV RNA revealed that the Korean EqHV isolates shared approximately 85.3–99.6% and 97.7–100% homology at the nucleotide and deduced amino acid level, respectively. Phylogenetic analysis revealed that the partial NS3 genes clustered with sequences previously reported as NPHV. Notably, sequences of EqHV detected in horses in the same region showed sequence divergence. The sequences of the polyprotein encoding region of two representative EqHVs shared 83.9% and 95.7% homology with each other at the nucleotide and deduced amino acid level, respectively. Comparison of the sequences of polyprotein encoding regions of Korean EqHV isolates and hepaciviruses from different hosts revealed that the NS3 and NS5B regions were most conserved among hepaciviruses. The results of the present study demonstrate that there is a high positive rate of EqHV in Korea and provide significant information regarding the geographical distribution and genetic variability of Korean EqHV isolates that will help improve global epidemiology of EqHV.     

海南岛斑点热疫源地的调查研究

本文报告在海南岛发现了斑点热疫源地，其人群北亚斑点热抗体的阳性率３８．３％，鼠类抗体阳性率５３．０％。从７种鼠检出抗体，其中优势种黄毛鼠（Ｒａｔｔｕｓｌｏｓｅａ）、黄胸鼠（Ｒ．ｆｌａｖｉｐｅｃｔｕｓ）和海南屋顶鼠（Ｒ．ｒａｔｔｕｓｈａｉｎａｎｉｃｕｓ）阳性率＞５０．０％，应用ＰＣＲ技术从该３种鼠检出斑点热群立克次体特异ＤＮＡ。小兽类寄生蜱３种、革螨５种，优势种分别为粒形硬蜱（Ｉｘｏｄｅｓｇｒａｎｕｌａｔｕｓ）和毒棘厉螨（Ｌａｅｌａｐｓｅｃｈｉｄｎｉｎｕｓ）；牛羊寄生蜱为微小牛蜱（Ｂｏｏｐｈｉｌｕｓｍｉｃｒｏｐｌｕｓ）。从粒形硬蜱和微小牛蜱检出斑点热群立克次体特异ＤＮＡ。     

Serological and molecular detection of Anaplasma phagocytophilum in Thoroughbred horses from Chilean racecourses

Anaplasma phagocytophilum is the causative agent of equine granulocytic anaplasmosis (EGA). This study aimed to perform serological and molecular surveys of A. phagocytophilum in thoroughbred horses from racecourses in Chile. Additionally, hematological findings related to A. phagocytophilum molecular positivity were addressed, and phylogenetic analysis of selected positive samples was performed. Complete blood count and msp2 gene real-time PCR were performed in 457 thoroughbred horses from three racecourses located in three different cities of Chile (Santiago, Viña del Mar and Concepción). Sera from horses in two racecourses (Santiago and Vina del Mar) were tested by Indirect fluorescent antibody test (IFAT) to detect IgG antibodies against A. phagocytophilum. The occurrence of A. phagocytophilum by real-time PCR was 13.6 % (62/457, 95 % CI: 10.8–16.3 %), with the highest occurrence observed in Santiago (26.5 %), followed by Concepción (9%), and the lowest in Viña del Mar (5%). The overall frequency of IgG antibodies to A. phagocytophilum was 7.9 % (23/290, 95 % CI: 4.8–12.7 %), with 9.9 % in Santiago and 6.5 % in Viña del Mar. Only three animals from Santiago Racecourse were positive in both real-time PCR and serology. PCR-positive horses from Santiago racecourse presented significantly lower hemoglobin, mean corpuscular value (MCV), and mean corpuscular hemoglobin concentration (CHCM), and higher eosinophil counts. Phylogenetic analysis based on the msp2 gene showed that A. phagocytophilum sequences found in the present study were closely related with A. phagocytophilum sequences from the USA and Europe. Anaplasma phagocytophilum DNA is detected for the first time in Chile.     

2011年厦门市某医院十例无菌性脑炎暴发病例病原学分析

目的 对2011年5月11-17日厦门某医院收治的10例无菌性脑炎患者进行病原学鉴定.方法 采集10例患者的咽拭子、肛拭子及部分患者的脑脊液,进行总肠道病毒核酸检测.总肠道病毒核酸阳性的样本,分别用肠道病毒A、B、C基因型各自的VP1通用引物扩增,VP1区扩增阳性的标本测序后进行序列分析.同时,用Vero细胞进行病毒分离培养,对培养成功的病毒与原始临床标本进行VP1序列同源性分析,从而对引发本次脑炎的病原进行鉴定.结果 10例脑炎患者中,有7例患者的临床标本总肠道病毒核酸检测阳性.这7例患者的咽拭子和(或)肛拭子标本肠道病毒B基因型VP1通用引物扩增阳性,测序后分析表明均为肠道病毒B基因型埃可病毒30(Echo 30),且与浙江2004年流行毒株的同源性达95.3%～97.1%.其中编号为XM2、XM3、XM4、XM8的咽拭子和XM3、XM6的肛拭子相互间序列同源性为99.4%～100.0%,但与XM1的咽拭子序列存在差异,同源性为92.8%～93.4%.此外,XM1、XM2、XM3、XM4、XM8咽拭子标本Vero细胞分离培养病毒成功,其VP1区部分序列与相应的原始标本同源性大于99.9%.结论 导致这次无菌性脑炎的病原体为肠道病毒B基因型Echo 30,并提示可能存在多种不同遗传背景的Echo 30在流行.

Abstract：

Objective To identify the etiology of an aseptic encephalitis outbreak (ten cases) in a hospital of Xiamen city from 11 to 17 May, 2011.Methods A total of ten patients′ throat swabs, anal swabs and cerebrospinal fluid were collected and detected by RT-PCR for pan-enterovirus. The samples containing detectable pan-enterovirus were tested by PCR with genotype-specific general primers located in VP1 region of enterovirus genotype A, B and C (HEV-A, B and C). The PCR products of VP1 segment were purified and sequenced, and phylogenetic analysis was performed. Meanwhile, the pathogens in those samples were isolated in Vero cell culture. Homologous analysis of VP1 sequences were carried out for the cultured virus samples and the original clinical samples to identify the outbreak etiology.Results Among the ten cases, seven cases were positive for pan-enterovirus nucleic acid. When tested by genotype-specific PCR, the throat and anal swab samples from those 7 patients were positive with HEV-B VP1 primers. Meanwhile, the HEV-B VP1 segments were sequenced and phylogenetic analyzed, which indicated the seven cases were all infected by enterovirus Echo 30. The sequences from those samples had homology of 95.3%-97.1% with the epidemic strains in Zhejiang, 2004. Out of the seven cases, the sequences of XM2,XM3,XM4,XM8 throat swab samples and XM3,XM6 throat samples showed 99.4%-100.0% homology which were different from the sequence of XM1, and the homology was 92.8%-93.4%. Furthermore, the viruses were isolated using Vero cells from XM1,XM2,XM3,XM4 and XM8 throat swab samples,and the VP1 sequence showed more than 99.9% homology with the original specimens.Conclusion The local outbreak of aseptic encephalitis was caused by Echo 30 of enterovirus genotype B, and the epidemic strains may have different genetic background.     

Rickettsia felis in Ctenocephalides spp. fleas, Brazil

In June 2000, suspected cases of Brazilian spotted fever (BSF) occurred in Coronel Fabriciano Municipality, Minas Gerais State, Brazil. Pooled fleas collected near two fatal cases contained rickettsial DNA. The nucleotide sequence alignment of the 391-bp segment of the 17-kDa protein gene showed that the products were identical to each other and to the R. felis 17-kDa gene, confirming circulation of R. felis in Brazil.     

Immunization with recombinant modified vaccinia Ankara (rMVA) constructs encoding the HA or NP gene protects ponies from equine influenza virus challenge

Two novel recombinant strains of modified vaccinia Ankara (rMVA) for the vaccination of horses against equine influenza virus were developed, and preliminary evidence of their immunogenicity in ponies was demonstrated [Breathnach CC, Rudersdorf R, Lunn DP. Use of recombinant modified vaccinia Ankara vectors for equine influenza vaccination. Vet Immunol Immunopathol 2004:98;127-36]. The present study assessed the protective efficacy of these rMVA strains in ponies, examined the advantage of combining rMVA vaccination with a DNA priming dose, and investigated the protection resulting from equine influenza nucleoprotein (NP) versus haemagglutinin (HA) vaccination. Twenty yearling ponies, seronegative for equine influenza virus, were divided into four groups of five. Group 1 and Group 2 ponies were vaccinated using a DNA prime-rMVA boost vaccination regimen, with HA- or NP-expressing vectors, respectively. Group 3 ponies were vaccinated with rMVA-HA only. Group 4 ponies served as unvaccinated controls. Vaccines were administered on days 0, 42 and 70, and all ponies were challenge infected with influenza virus on day 100. Antigen-specific antibody and cellular immune responses to each vaccination regimen were monitored throughout the experiment. Both groups of HA-vaccinated ponies were significantly protected from clinical disease following challenge infection, demonstrating the efficacy of rMVA vaccination with or without a DNA prime. NP-vaccination provided more limited protection from clinical disease. The protective post-vaccinal immune responses were characterized by antigen-specific IgGa, IgGb and IgA antibodies which were induced both in serum and in nasal secretions. Virus-specific lymphoproliferative and IFN-gamma mRNA responses were also elicited by each vaccination regimen. These data demonstrate that vaccination of horses with rMVA alone, or as part of a prime-boost regimen, is an effective means of inducing protective immunity to influenza virus infection, and also indicate that NP-specific immune responses can contribute to protection of horses.     

Equine interferon gamma synthesis in lymphocytes after in vivo infection and in vitro stimulation with EHV-1

Equine cytotoxic T lymphocyte (CTL) responses to equine herpesvirus-1 (EHV-1) are well characterised but little is known about the cytokine response after infection or vaccination. EHV-1 is common in horses and infects lymphocytes in vivo. This virus was used as a model to measure the synthesis of interferon gamma (IFN-gamma) by equine peripheral blood mononuclear cells (PBMC) after in vivo infection and/or in vitro stimulation with EHV-1. Both flow cytometry and ELISPOT assays were used to quantify equine IFN-gamma using a mouse anti-bovine IFN-gamma monoclonal antibody (clone CC302; shown to cross-react with recombinant equine IFN-gamma) and a rabbit anti-canine IFN-gamma polyclonal antibody. The percentage of PBMC synthesising IFN-gamma after in vitro stimulation with EHV-1 increased with age. In yearlings infected experimentally with EHV-1, PBMC showed two peaks of IFN-gamma synthesis, 11 and 56 days after infection. The IFN-gamma synthesis was principally associated with CD8(+) cells. The patterns of IFN-gamma synthesis detected by intracellular IFN-gamma staining or ELISPOT were compared with CTL data and shown to be similar. These methods were also applied successfully to frozen samples of PBMC. Measurement of equine IFN-gamma using these simple techniques can now be applied to future studies on protective cellular immune responses following virus infection and/or vaccination of horses.     

云南省保山市一起地方性斑疹伤寒暴发的调查

目的 了解云南省保山市一起地方性斑疹伤寒暴发的流行病学特征.方法 收集患者的流行病学资料.应用外斐反应和间接免疫荧光方法同时检测患者血清中莫氏立克次体、恙虫病东方体IgG抗体.用PCR检测鼠类脾脏标本中莫氏立克次体、普氏立克次体gltA基因,恙虫病东方体56kDa蛋白基因,斑点热群立克次体ompA基因,埃立克体16S rRNA基因和无形体16SrRNA基因.结果 2009年7-8月保山市隆阳区该起地方性斑疹伤寒累计发病58例,其中48例为临床诊断病例,10例为实验室诊断病例.3例地方性斑疹伤寒实验室诊断病例存在Karp型恙虫病东方体感染.PCR检测黄胸鼠脾脏85份,其中莫氏立克次体gltA片段阳性3份(阳性率为3.5%),其序列与莫氏立克次体Wilmington株(GenBank U59714.1)同源性为100%;普氏立克次体、恙虫病东方体、斑点热群立克次体、无形体和埃立克体均为阴性.结论 经流行病学、临床资料和实验室检测证实,保山市隆阳区该起疫情为地方性斑疹伤寒.从当地优势鼠种黄胸鼠中检测到莫氏立克次体核酸及序列,表明当地存在地方性斑疹伤寒疫源地.

Abstract：

Objective To understand the epidemiologic characteristics of endemic typhus in Baoshan city. Methods Epidemiological data were collected and characteristics were analyzed. IgG antibody(Ab) of Rickettsia mooseri and Orientia tsutsuganushi in serum of patients were tested using both Weil-Felix and IFA method. The Rickettsia mooseri gltA gene, Rickettsia prowazekii gltA gene,Orientia tsutsugamushi 56 kDa protein gene, SFGR ompA gene, Ehrlichia sp. 16S rRNA gene and Anaplasma sp. 16S rRNA gene in spleen of mice were examined by PCR. Results Fifty- eight endemic typhus cases were found in Longyang district of Baoshan city, during July to August, 2009.Among them, 48 cases were confirmed by clinical diagnosis and 10 cases by laboratory tests. The Ab of Orientia tsutsugamushi Karp serotype was detected in 3 cases from laboratory diagnosis. The spleen samples from 85 Rattns flavipectus were tested using PCR. Of them, 3 samples for Rickettsia mooseri gltA gene showed positive (positive rate was 3.5% ), and the homology of 3 Rickettsia mooseri and Rickettsia mooseri Wilmington strain (GenBank U59714.1) was 100% through comparing gene sequence. The results of PCR for detecting Rickettsia prowazekii, Orientia tsutsugamushi, SFGR,Anaplasma sp. and Ehrlichia. sp were all negative. Conclusion The outbreak of endemic typhus was confirmed in Longyang district of Baoshan city through epidemiological data, clinical diagnosis and laboratory tests. Rickettsia mooseri DNA was detected in the dominant Raw flavipectus, suggesting that endemic typhus did exist in the local areas.     

Importance of equine piroplasmosis antibody presence in Spanish horses prior to export

Serological analysis of equine piroplasmosis (EP), caused by Theileria equi and Babesia caballi, is included in the export testing requirements for most of the countries worldwide, thus involving a high economic impact on equine industry of EP-endemic countries, such as Spain. A total of 3368 serum samples from healthy horses collected prior to export between 2015 and 2018 in Spain were tested for antibodies against T. equi and B. caballi by using a competitive inhibition enzyme-linked immunosorbent assay (cELISA). The overall seroprevalence results in Spain revealed that almost a quarter of the horses analysed (24.1 %; 95% CI 22.6–25.5) could not be exported to countries free from EP. The implementation of prevention measures such as the use of acaricides and daily checks for ticks in horses, as well as regular serological screening of horses in Spain would aid to increase the number of horses exported to other countries.     

Giardia duodenalis sub-Assemblage of animal and human origin in horses

In order to evaluate infection occurrence and the potential zoonotic role of horse isolates of Giardia duodenalis, 431 individual fecal samples were genetically characterized by PCR tests –coupled sequencing and phylogenetic analysis. Thirty-seven (8.6%) animals resulted infected by different Assemblage. The presence of sub-Assemblage was assessed by characterizing the β-giardin gene for 16 of the 37 positive horses. Ten isolates showed 99.6% to 100% homology with the sub-Assemblage described as B1–2 and B1–6, three Assemblage A showed 99.8% homology with sub-Assemblage A1, while one Assemblage E displayed 98.8% homology with sub-Assemblage E3. Furthermore, one isolate characterized as Assemblage A showed 99.6% homology with the sub-Assemblage B1–2 and one characterized as E was 100% identical with sub-Assemblage B1–6. These results demonstrate the presence of both animal and human sub-Assemblage of G. duodenalis in horses from Italy. Epidemiological and sanitary implications are discussed.     

Non-primate hepacivirus transmission and prevalence: Novel findings of virus circulation in horses and dogs in Morocco

Non-primate hepacivirus (NPHV) is a homolog of hepatitis C virus and has been isolated from dogs and horses. Data on NPHV prevalence and distribution are not complete, and there is a particular lack of reports from the African continent. The present study represents the first investigation of NPHV prevalence in horses and dogs in North Africa.Blood was collected from 172 horses and 36 dogs at different locations in Morocco, and screened for NPHV RNA using nested PCR targeting 5’UTR and NS3 regions and analyzed for anti-NPHV NS3 antibody using a Gaussia luciferase immunoprecipitation system—to determine seroprevalence. Eight sequences of the NS3 region isolated from positive serum samples were targeted for phylogenetic analysis.Horses and dogs showed respective NPHV RNA positivity rates of 10.5% and 5.5%, and seroprevalences of 65.7% and 8.33%. Juvenile horses appeared more susceptible to infection, with a 23.5% NHPV RNA positivity rate. Seropositivity was more extensive in mares than stallions (77.14% vs. 46.27%, p < 0.0001). Phylogenetically, that NPHV NS3 genes isolated from horses and dog are clustered together. The NPHV strains we detected showed no correlation with geographic location within Morocco.In conclusion, Moroccan horses showed much evidence of previous and/or current NPHV infection, with young age and female sex as noted potential risk factors. Interestingly, NPHV is circulating in dogs as well as horses, suggesting that it has crossed species barriers and that horses and dogs are potential vectors by which an ancestor to hepatitis C virus was transmitted into human populations.