Characterization of pigeon paramyxoviruses (Newcastle disease virus) isolated in South Africa from 2001 to 2006

Pigeon paramyxovirus type 1 (PPMV-1), a variant of Newcastle disease virus that primarily affects doves and pigeons has been isolated in South Africa since the mid-1980s. Phylogenetic evidence indicates that pigeon paramyxovirus type 1 viruses were introduced into South Africa on multiple occasions, based on the presence of two separate lineages, 4bi and 4bii, that have been circulating in Europe and the Far East since the early 1990s. During 2006, a PPMV-1 virus was isolated from an African ground hornbill (Bucorvus leadbeateri) which became acutely infected with PPMV-1 and died, probably after scavenging off infected dove carcasses in the region, since a closely-related PPMV-1 strain was also isolated from doves collected nearby. The hornbill isolate had ICPI and MDT values characteristic of PPMV-1 strains. The threat of PPMV-1 to poultry production and biodiversity in southern Africa highlights the importance of monitoring the spread of this strain.     

Sequence and phylogenetic analysis of the non-structural 3A and 3B protein-coding regions of foot-and-mouth disease virus subtype A Iran 05

The A Iran 05 foot-and-mouth disease virus (FMDV) subtype was detected in Iran during 2005 and has proven to be highly virulent. This study was undertaken to focus on molecular and phylogenetic analysis of 3A and 3B coding-regions in the A Iran 05 field isolate. To assess the genetic relatedness of A Iran 05 isolate the nucleotide and predicted amino acid sequences of the 3AB region of type A FMDV isolates were compared with twenty previously described type A FMDV isolates. The phylogenetic tree based on the 672 bp 3AB gene sequences of type A FMDV from thirteen different locations clustered them into five distinct lineages. The A Iran 05 isolate clustered in lineage A along with four type A variants and was closely matched with viruses isolated in Turkey and Pakistan during 2005~2006. The number of protein sequence differences exhibited by each of the isolates revealed that A Iran 05 isolate contains three amino acid substitutions at positions 47 and 119 of 3A and 27 of the 3B coding region. The nucleotide identity between A Iran 05 and the other four isolates of lineage A was estimated to be 98%.     

The fowl adenovirus (Fadv-11) outbreak in Iranian broiler chicken farms: The first full genome characterization and phylogenetic analysis

Fowl adenoviruses D and E (FAdV-D and E) can cause inclusion body hepatitis (IBH) in commercial chicken flocks. Recently, IBH outbreaks have been increasingly reported in different regions of Iran, particularly in broiler farms. The present study was conducted to perform, for the first time, a complete genome characterization of a FAdV isolate from an IBH outbreak in Iran. Briefly, liver samples were collected from affected broiler flocks and following viral DNA extraction and confirming by PCR technique; one positive sample was selected from an affected flock to conduct a complete genome sequencing. The current FAdV, named "Fowl_Adenovirus_D_isolate_iran/UT-Kiaee_2018", was placed into FAdV-11 serotype (D species). According to the complete genome sequence analysis, UT-Kiaee had high homology with Chinese and Canadian FAdV. The partial sequence of the hexon gene revealed that UT-Kiaee shared 100% identity with previous Iranian FAdVs. The present study was the first to report full genome FAdV in Iran and complete the puzzle of molecular epidemiology of FAdV in Iran through determining the possible origin of Iranian FAdvs, which are the causative agents of recent IBH outbreaks in Iran.     

Sequence and phylogenetic analysis of neuraminidase genes of H9N2 avian influenza viruses isolated from commercial broiler chicken in Iran (2008 and 2009)

A total of 512 tissue samples collected from 30 farms located in various states of Iran during 2008–2009 as part of a program to monitor avian influenza viruses (AIVs) infection in Iran’s poultry population. To determine the genetic relationship of Iranian viruses, neuraminidase (NA) genes from ten isolates of H9N2 viruses isolated from commercial chickens in Iran during 2008–2009 were amplified and sequenced. The viruses’ neuraminidase gene was >90% similar to those of A/Quail/Hong Kong/G1/97 (H9N2) sublineage. The neuraminidase stalk regions in these Viruses had no deletion as compared to that of chicken/Beijing/1/94 sublineage (Beijing-like viruses) and the two human isolates A/HK/1073/99, A/HK/1074/99. Phylogenetic analysis of neuraminidase (NA) gene showed that it shares a common ancestor A/Quail/Hong Kong/G1/97 isolate which had contributed the internal genes of the H5N1 virus. The results of this study indicated that No (Beijing-like) virus and (Korean-like) virus were found in chickens in Iran, and the NA genes of H9N2 influenza viruses circulating in Iran during the past years were well conserved and the earlier Iranian isolates may be considered to represent such a progenitor.     

猪流感病毒H1N1分离株HA基因的克隆与序列分析

无锡某猪场发生不同程度的猪呼吸系统疾病.对发病猪采集鼻拭子,经双抗处理后,接种鸡胚,收集24 h后死亡的鸡胚尿囊液.对一株具有血凝性的病毒分离株进行RT-PCR鉴别,结果为H1亚型流感病毒.利用RT-PCR扩增尿囊液中的流感病毒HA基因.经pGEM-T easy载体克隆、序列测定和进化树分析,结果表明,在该猪群中分离获得的该株SIV为古典型H1N1.     

Genotyping of avian infectious bronchitis virus in Iran: Detection of D274 and changing in the genotypes rate

The coronavirus avian Infectious bronchitis virus (IBV) poses economic threats to poultry farms worldwide, affecting the performance of both meat-type and egg-laying birds. To define the evolution of recent IBVs in Iran, a genetic analysis based on hypervariable nucleotide sequences of S1 gene was carried out. Tracheal swab samples were collected from 170 Broiler flocks during 2017. Ten tracheal swabs from each flock pooled. From a total number of 170 flocks tested, 84.71% found to be positive. Phylogenetic tree analysis revealed the presence of D274 as a first time in Iran. IS/1494/06 was showed to be dominant IBV type circulating in broiler farms with a significantly higher prevalence than other four genotypes. Considering fluctuations in QX-type prevalence in recent years, continuous monitoring is necessary to reduce economic consequences in layer and broiler farms. The findings highlight the importance of using modified vaccination strategies that are adapted to the changing disease scenario.     

Molecular characterization of a 13-amino acid deletion in VP1 (1D) protein and novel amino acid substitutions in 3D polymerase protein of foot and mouth disease virus subtype A/Iran87

The nucleotide sequence of the VP1 (1D) and partial 3D polymerase (3D^pol) coding regions of the foot and mouth disease virus (FMDV) vaccine strain A/Iran87, a highly passaged isolate (~150 passages), was determined and aligned with previously published FMDV serotype A sequences. Overall analysis of the amino acid substitutions revealed that the partial 3D^pol coding region contained four amino acid alterations. Amino acid sequence comparison of the VP1 coding region of the field isolates revealed deletions in the highly passaged Iranian isolate (A/Iran87). The prominent G-H loop of the FMDV VP1 protein contains the conserved arginine-glycine-aspartic acid (RGD) tripeptide, which is a well-known ligand for a specific cell surface integrin. Despite losing the RGD sequence of the VP1 protein and an Asp₂₆→Glu substitution in a beta sheet located within a small groove of the 3D^pol protein, the virus grew in BHK 21 suspension cell cultures. Since this strain has been used as a vaccine strain, it may be inferred that the RGD deletion has no critical role in virus attachment to the cell during the initiation of infection. It is probable that this FMDV subtype can utilize other pathways for cell attachment.     

斑点叉尾鮰源致病性维氏气单胞菌的分离与生物学特性鉴定

为鉴定一起致斑点叉尾鮰突然发病死亡的病原，本研究从发病斑点叉尾鮰中分离到1株致病菌GZTL2017，通过临床解剖观察、细菌分离培养、革兰氏染色镜检、动物回归试验、生化试验、16S rDNA序列分析、部分毒力基因检测和药敏试验进行鉴定。临床解剖观察结果显示，患病斑点叉尾鮰呈现体表溃疡、鳍根部出血、烂腮、肠管充血等症状；分离菌在培养基中呈现表面湿润凸起、边缘光滑半透明、形态均一的乳白色菌落；革兰氏染色镜检显示，分离菌为单个或成双存在、两端钝圆的短直阴性杆菌；动物回归试验显示，该分离菌有较强的致病性；生化试验结果显示，分离菌具有运动性，且氧化酶、V-P、赖氨酸脱氢酶等反应阳性，精氨酸双水解酶、硫化氢等反应阴性；16S rDNA基因序列系统进化树显示，该菌与维氏气单胞菌聚为一支，同源性均>99%；毒力基因检测结果显示，该菌能检出气溶素基因（Aer）、黏附素基因（Aha）、外膜蛋白基因（OmpA）3种毒力基因；药敏试验结果显示，该菌对氟苯尼考、强力霉素、氧氟沙星等14种药物敏感；对诺氟沙星、新霉素、万古霉素等7种药物中度敏感；对麦迪霉素、苯唑西林、头孢拉定等8种药物耐药，且对氟苯尼考、强力霉素的药物最小抑菌浓度分别为1和2 μg/mL。本试验成功分离到1株维氏气单胞菌，为斑点叉尾鮰维氏气单胞菌病防治提供参考依据。     

Survey for Haemoproteus spp., Trichomonas gallinae, Chlamydophila psittaci, and Salmonella spp. in Galapagos Islands columbiformes.

Endemic free-ranging Galapagos doves (Zenaida galapagoensis) and introduced rock doves (Columba livia) were surveyed in several islands of the Galapagos archipelago to establish sample prevalence of hemoparasites, Trichomonas gallinae, Chlamydophila psittaci, and Salmonella species. A Haemoproteus sp., the only hemoparasite identified, was found in 89% of the Galapagos doves sampled but not in the rock doves. Trichomonas gallinae was detected by polymerase chain reaction in 44% of rock doves from San Cristobal but in none of the Galapagos doves. Chlamydophila psittaci was detected from cloacal swabs in 6% of the Galapagos doves but in none of the rock doves sampled. All positive cases of C. psittaci occurred on Espa?ola, where the crude sample prevalence was 24%. A polymerase chain reaction-based Salmonella test failed to show evidence of this organism from any birds sampled.     

广西地区猪肠病毒G型的流行病学调查及VP1基因序列分析

为分析广西地区猪肠病毒G型（EV-G）的流行状况及分子遗传演化特征,本研究收集2017－2018年广西地区222份临床腹泻样品,进行EV-G的检测、流行病学调查及VP1基因的扩增、克隆和序列分析。流行病学调查结果显示,广西地区2017－2018年EV-G的样品阳性率为6.76%（15/222）,而猪场阳性率则高达16.98%（9/53）。值得注意的是,广西地区2017－2018年EV-G的样品阳性率从2017年的4.55%上升至2018年的10.00%,上升幅度较大。同时发现该病毒在冬春季节的检出率较高。序列同源性分析结果显示,本研究获得的2株EV-G VP1基因之间的核苷酸同源性为99.2%,氨基酸同源性为98.8%,与国内外毒株VP1基因的核苷酸同源性为62.4%~80.1%,氨基酸同源性为45.7%~71.7%。遗传进化分析显示,本研究获得的2株EV-G VP1序列在同一分支,均属于G1亚型,与美国分离株13-03212遗传关系密切。氨基酸序列比较结果显示,广西地区该2株EV-G VP1氨基酸在多个位点发生了变异。抗原指数分析显示,广西地区EV-G型流行毒株的抗原性变化较大。表明2017－2018年广西地区存在一定程度的EV-G感染,本研究结果为明确EV-G型的流行概况及EV-G型的分子特性提供了参考依据。     

Full genome characterization of Iranian H5N8 highly pathogenic avian influenza virus from Hooded Crow (Corvus cornix), 2017: The first report

During 2014–2017 Clade 2.3.4.4 H5N8 highly pathogenic avian influenza viruses (HPAIVs) have spread worldwide. In 2016, an epidemic of HPAIV H5N8 in Iran caused mass deaths among wild birds, and several commercial poultry farms and captive bird holdings were affected and continue to experience problems. Several outbreaks were reported in 2017. One of them is related to Hooded crow (Corvus cornix) in a national park in Esfahan province in 2017. Whole genome sequencing and characterization have been done on the detected H5N8 sample. Based on HA sequencing results, it belongs to 2.3.4.4 clade, and the cleavage site is (PLREKRRKR/G). Phylogenetic analysis of the HA gene showed that the Iran 2017 H5N8 virus clustered within subgroup Russia 2016 2.3.4.4 b of group B in H5 clade 2.3.4.4 HPAIV.On the other hand, the NA gene of the virus is placed in group C of Eurasian lineage. Complete genome characterization of this virus revealed probable reassortment of the virus with East-Asian low-pathogenic influenza viruses. Furthermore, the virus possessed some phenotypic markers related to the increased potential for transmission and pathogenicity to mammals at internal segments. This study is the first full genome characterization H5N8 HPAIV in Iran. The data complete the puzzle of molecular epidemiology of H5N8 HPAIV in Iran and the region. Our study provides evidence for fast and continuing reassortment of H5 clade 2.3.4.4 viruses, that might lead to changes in virus structural and functional characteristics such as the route and method of transmission of the virus and virus infective, pathogenic and zoonotic potential.     

Trichomonosis in free-ranging Eurasian collared doves (Streptopelia decaocto) and African collared dove hybrids (Streptopelia risoria) in the Caribbean and description of ITS-1 region genotypes

We report the first documented occurrence of an outbreak of trichomonosis in a free-ranging small flock of Eurasian collared doves (Streptopelia decaocto) and African collared dove hybrids (Streptopelia risoria) in the Caribbean. In total, 18 birds were examined, including six African collared dove x Eurasian collared dove hybrids and 12 Eurasian collared doves. The affected age class consisted of adults. Sex distribution was equal. With a flock population size of 200 birds, mortality rate for the outbreak was estimated at 15-20%. Living birds were weak, showing evidence of mucus-stained beaks and open-mouth breathing. Caseous ulcerative yellow lesions were restricted to the upper gastrointestinal tract, with the exception of one bird, which had lesions in the upper gastrointestinal tract and in the liver. Ninety-four percent (17/18) of the affected birds had multiple extensive lesions. Lesions located on the roof of the oral cavity extended in 33% (6/18) into the orbit and in 11% (2/18) into the braincase. Using wet-mount microscopy, we were able to confirm Trichomonas gallinae in 22% (4/18) of the sampled animals. Fifteen samples submitted for PCR analysis tested positive. Sequence analysis of the internal transcribed spacer 1 (ITS-1) region of the ribosomal RNA (rRNA) revealed two distinct genotypes of Trichomonas. One sequence had 100% identity to the prototype T. gallinae isolate, whereas the other sequences had 98-100% identity to recently described Trichomonas-like parabasalid. On the basis of gross and histologic findings, along with the sequence results from the columbids in this report, it is likely that this Trichomonas-like parabasalid is pathogenic.     

猪流行性腹泻病毒JS2017株的分离鉴定及S基因序列分析

为了解江苏省某规模化猪场猪流行性腹泻病毒(PEDV)遗传变异情况,本试验采集腹泻仔猪小肠组织及肠道内容物,通过RT-PCR进行PEDV鉴定。将检测为阳性的病料组织经20μg/mL胰酶处理后,接种到Vero细胞中进行培养,将细胞培养物进行反复冻融后,收获病毒液,进行RT-PCR鉴定。采用分段重叠策略对其S基因进行全基因序列扩增,并在此基础上分析该毒株S基因的核苷酸同源性及遗传进化关系。结果显示,经RT-PCR鉴定,病料组织呈PEDV阳性,阳性病料接种Vero细胞盲传4代后,出现明显的细胞病变,表现为细胞合胞体病变,空泡化,最终裂解脱落。收获的病毒液经RT-PCR鉴定后,确认分离到1株PEDV,命名为JS2017株。核苷酸序列分析表明,JS2017株S基因序列与美国变异株OH1414、加拿大变异株ON-007、中国变异株PEDV-LYG和CH/PDS/2015位于同一进化分支,其核苷酸同源性高达99.0%以上;其中与中国变异株YC2014亲缘关系最近,核苷酸同源性为100.0%;与经典株CV777、DR13株同源性较低,亲缘关系较远。结果表明,本研究分离获得的JS2017毒株是一株PEDV地方流行变异毒株,与2014年中国江苏分离株(YC2014株)亲缘关系最密切,与当前猪流行性腹泻疫苗株亲缘关系较远。     

猪流感病毒广东株分离鉴定及遗传进化分析

本研究从广东省某猪场采集37份疑似猪流感症状的猪鼻拭子样品,接种于9日龄SPF鸡胚并收集尿囊液,通过血凝试验、血凝抑制试验和RT-PCR鉴定,分离得到一株猪流感病毒,经RT-PCR分别扩增8个基因片段,进行基因测序及序列分析,与GenBank收录的参考毒株比对并构建进化树。结果显示,分离毒株为H1N1亚型猪流感病毒,将其命名为A/swine/Guangdong/2/2018（H1N1）。遗传进化分析显示,分离株8个片段的核酸序列与A/swine/Guangdong/L3/2009（H1N1）对应序列的同源性均达99%以上,与经典型H1N1亚型猪流感病毒处于同一分支。分离毒株HA的裂解位点为PSIQSR↓GL,符合低致病性流感病毒分子特征。HA基因受体位点为190D、225G和226Q,表明本毒株既可以结合SAα-2,6-Gal型人类流感病毒SA受体,也有结合SAα-2,3-Gal型禽类流感病毒SA受体的可能,在28、40、104、304、498、557位氨基酸处有6个潜在糖基化位点;NA蛋白在50、58、63、68、98、146、235位氨基酸处有6个潜在糖基化位点,NA蛋白氨基酸序列活性中心位点为119E、199D、223I、275H、293R、295N,氨基酸分析位点未出现突变,表明本分离株对神经氨酸酶抑制剂类药物的敏感性较高,但在M2蛋白中,31位氨基酸由敏感型的（S）突变为抗药的（N）,提示可能对金刚烷胺类药物产生耐药性。开展猪流感病毒分离鉴定与遗传进化分析将为广东地区的猪流感流行和变异情况提供重要信息。     

Venous blood gas and lactate values of mourning doves (Zenaida macroura), boat-tailed grackles (Quiscalus major), and house sparrows (Passer domesticus) after capture by mist net, banding, and venipuncture

Blood gas partial pressures, pH, and bicarbonate and lactate concentrations were measured from the basilic vein of mourning doves (Zenaida macroura) and the jugular vein of boat-tailed grackles (Quiscalus major) and house sparrows (Passer domesticus) to assess immediate impacts of mist net capture and handling for banding and venipuncture. Mourning doves and house sparrows exhibited mild acidemia (median [minimum-maximum] venous blood pH(41 degrees C) = 7.394 [7.230-7.496] and 7.395 [7.248-7.458], respectively), relative to boat-tailed grackles (Quiscalus major; 7.452 [7.364-7.512]), but for different reasons. Mourning doves exhibited relative metabolic acidosis (lower venous blood pH, higher lactate concentrations, lower bicarbonate, and no significant differences in partial pressure of CO2 (pCO2) or partial pressure of O2 (pO2) compared with boat-tailed grackles). House sparrows exhibited relative respiratory acidosis (lower venous blood pH, higher pCO2, lower pO2, and no significant differences in bicarbonate and lactate concentrations compared with boat-tailed grackles). All birds captured by mist net and handled for banding and venipuncture experienced some degree of lactic acidemia; and values were greater in mourning doves (lactate, 7.72 [3.94-14.14] mmol/L) than in boat-tailed grackles (5.74 [3.09-8.75] mmol/L) and house sparrows (4.77 [2.66-12.03] mmol/L), despite mourning doves resisting least and being easiest to disentangle from the mist net. House sparrows were more susceptible to respiratory acidosis, warranting particular care in handling birds <30 g to minimize interference with ventilation. The different sample collection site for mourning doves may have affected results in comparison with the other two species, due to activity of the wing muscles. However, despite the higher lactate concentrations, pCO2 was relatively low in doves. The metabolic, respiratory, and acid-base alterations observed in this study were minor in most cases, indicative of the general safety of these important field ornithology techniques. The effect of other adverse conditions, however, could be additive.     

1株鸭源非O1/O139群霍乱弧菌的分离鉴定与致病性分析

为探明引起贵州省某鸭场雏鸭发病的病原及其致病性和耐药情况，本研究对该鸭场送的疑似细菌感染病鸭进行剖检，取鼻黏膜、心脏和肝脏等组织器官接种于培养基中进行细菌分离鉴定，通过对分离菌进行药敏试验、动物回归试验和毒力基因检测研究其耐药情况和致病性。结果显示，分离菌在血琼脂培养基上生长16 h后呈现为边缘整齐、有光泽的乳白色菌落，伴有β-溶血现象，经革兰氏染色后在生物显微镜下呈两端钝圆、弧状、排列无规则的革兰氏阴性短小杆菌，与霍乱弧菌相符；16S rDNA基因序列同源性及系统进化树显示，该分离菌与霍乱弧菌同源性高达99.6%~99.7%聚为一支；药敏试验结果显示，分离菌对大部分药物都表现为耐药，其中对氨苄西林、克林霉素、复方新诺明、苯唑西林和克林霉素等抗菌药耐药性较强，对头孢哌酮和头孢曲松敏感；动物回归试验显示，分离菌可导致试验组雏鸭5 d内全部发病死亡，表明该分离菌对雏鸭具有较强的致病性；毒力基因PCR检测结果显示，检测的霍乱弧菌相关毒力基因hlyA、ompW和chxA为阳性，而检测的O1群rfb、O139群rfb、tcpA及ctxA基因为阴性，表明本次分离的霍乱弧菌携带有致病基因，但不属于O1和O139血清群。结果表明，该鸭场雏鸭发病的疫情病原为非O1/O139血清群霍乱弧菌，该菌致病性强且对多种抗菌药物耐药。本试验结果为贵州省鸭霍乱弧菌病的防控提供了参考依据。     

一株野鸟源重组禽流感(H6N1)病毒基因组分子特征分析

2017年在江苏省野生豆雁粪便中分离得到1株H6N1亚型禽流感病毒A/Anser fabalis/Jiangsu/J746/2017(H6N1)(J746)。本研究对J746进行了全基因组测序,并对其进行了遗传进化分析。遗传进化分析结果表明:与HA和NA基因同源性最高的毒株为A/wild waterfowl/Korea/F14-5/2016(H6N1),同源性为99.4%。HA基因与流行于韩国、日本和孟加拉的N1、N2、N8亚型毒株处于同一分支,NA基因与韩国野生水禽的H6、H7亚型毒株处于同一分支,PB2基因与中亚及东亚地区低致性病毒株处于同一分支,PB1基因和NP基因与流行在东南亚的低致病性毒株处于同一分支,PA基因和M基因均处于欧亚分支,但PA形成了独立的小分支,NS基因与分离于中国中南部和日本的毒株聚集在一起。氨基酸位点分析表明,神经氨酸(NA)蛋白存在H274Y突变,该突变可增强病毒对神经氨酸酶抑制剂药物的耐药性;同时在PB2蛋白中发现与增强对小鼠致病性有关的L89V突变,在NS1蛋白中发现与增强对小鼠致病性、提高复制能力和改变宿主嗜性有关的P42S、L103F、I106M、N205S突变。综上所述,J746毒株基因组构成来源复杂,是由多个国家和地区形成的一株多元重组病毒。     

洱源、怒江猫源弓形虫虫株的分离与鉴定

为了分离猫源弓形虫,本试验从云南洱源、怒江两地区捕捉18只野猫,取其心脏、肝脏、肺脏、脑组织用盐酸—胃蛋白酶溶液消化处理后,腹腔接种小白鼠,将分离到的弓形虫虫株至少传3代,用特异PCR方法对所分离的虫株进行鉴定。结果表明,从18只野猫的样品中分离出3株弓形虫虫株,用特异性引物对3株虫株进行PCR鉴定,均得到弓形虫的特异性目的条带,测序结果表明所扩增出的DNA片段确为弓形虫核糖体B1基因部分序列。同源性比对分析结果显示分离株与T.gondii B1的同源性为100.0%。将动物组织用盐酸—胃蛋白酶溶液消化处理后腹腔接种小白鼠是一种分离弓形虫虫株较理想的方法,对弓形虫B1基因进行特异性扩增,可以快速地鉴定弓形虫虫株。     

Isolation and Identification of Toxoplasma gondii Isolates from Cats in Eryuan and Nujiang

The assay was aimed to isolate Toxoplasma gondii (T.gondii) strains from stray cat in Eryuan and Nujiang of Yunnan province.The cat tissues (heart,liver,lung and brain) were digested by acid pepsin solution,intraperitoneally inoculated in Kunming mice,passaged at least 3 generations,and followed by specific PCR amplification of partial B1 gene using species-specific primers.Three T.gondii isolates were isolated from 18 stray cats,PCR result showed that we got the specific target band,and the sequence result of the specific PCR product showed that it was ribosome B1 gene sequence of T.gondii. Homology comparison analysis showed that the isolates was 100.0% homology with T.gondii B1.The method of inoculation into the mice with the tissues that was digested by acid pepsin solution was an effective way to isolate T.gondii strain from animals,and the specific PCR assay was an accurate method for the rapid identification of T.gondii.     

Molecular characterization of Malaysian fowl adenovirus (FAdV) serotype 8b species E and pathogenicity of the virus in specific-pathogen-free chicken

BackgroundInclusion body hepatitis (IBH) is an economically important viral disease primarily affecting broiler and breeder chickens. All 12 serotypes of fowl adenovirus (FAdV) can cause IBH.ObjectivesTo characterize FAdV isolates based on phylogenetic analysis, and to study the pathogenicity of FAdV-8b in specific-pathogen-free (SPF) chickens following virus inoculation via oral and intramuscular (IM) routes.MethodsSuspected organ samples were subjected to virus isolation and polymerase chain reaction (PCR) for FAdV detection. Hexon gene sequencing and phylogenetic analysis were performed on FAdV-positive samples for serotype identification. One FAdV-8b isolate, UPM/FAdV/420/2017, was selected for fiber gene characterization and pathogenicity study and was inoculated in SPF chickens via oral and IM routes.ResultsThe hexon gene phylogenetic analysis revealed that all isolates belonged to FAdV-8b. The fiber gene-based phylogenetic analysis of isolate UPM/FAdV/420/2017 supported the grouping of that isolate into FAdV species E. Pathogenicity study revealed that, chickens infected with UPM/FAdV/420/2017 via the IM route had higher clinical score values, higher percent mortality, higher degree of the liver lesions, higher antibody response (p < 0.05), and higher virus shedding amounts (p < 0.05) than those infected via the oral route. The highest virus copy numbers were detected in liver and gizzard.ConclusionsFAdV-8b is the dominant FAdV serotype in Malaysia, and pathogenicity study of the FAdV-8b isolate UPM/FAdV/420/2017 indicated its ability to induce IBH in young SPF chickens when infected via oral or IM routes.