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Genetic relationship of Salmonella isolates found in subcutaneous abscesses in leopard geckos (Eublepharis macularius)
2020
Zając, Magdalena | Maluta, Aleksandra | Wasyl, Dariusz | Skarżyńska, Magdalena | Lalak, Anna | Samcik, Ilona | Kwit, Renata | Szulowski, Krzysztof
The article describes the occurrence and phylogenetic relationship of Salmonella isolates found in subcutaneous abscesses of leopard geckos. The aim of the study was to determine the cause of the abscesses and to characterise isolated Salmonella strains. Samples of abscesses from five animals and internal organs (lungs, liver, and gut) of three of them were tested for Salmonella according to the PN-EN ISO 6579:2002/A1:2007 standard. The antimicrobial resistance was evaluated by minimal inhibitory concentrations and the genetic similarity of the isolates was assessed with pulsed field gel electrophoresis (PFGE). In total, seventeen Salmonella isolates belonging to five different serovars were found to be susceptible to all tested antimicrobials except streptomycin. The serovars were S. Hadar, S. Fluntern, S. Tennessee, S. enterica subsp. salamae 55:k:z₃₉, and S. Kentucky. Up to three serovars from different organs were isolated from the same individual. In two geckos, Salmonella were detected in the lungs. In three serovars, XbaI-PFGE typing revealed indistinguishable isolates from organs and abscesses. Multiple Salmonella serovars might be involved in abscess formation and infections. The occurrence of the same PFGE profiles of the isolates may testify to the role of opportunistic organisms in causing infection.
Show more [+] Less [-]Genetic characterisation of African swine fever virus in outbreaks in Ha Nam province, Red River Delta Region of Vietnam, and activity of antimicrobial products against virus infection in contaminated feed
2020
Tran, Ha Thi Thanh | Truong, Anh Duc | Ly, Duc Viet | Vũ, Thị Hảo | Hoàng, Văn Tuấn | Nguyễn, Thị Chính | Chu, Thi Nhu | Nguyen, Thi Huyen | Pham, Ngoc Thi | Nguyen, Tinh | Yersin, Andrew G. | Dang, Hoang Vu
African swine fever (ASF) was officially reported in Vietnam in February 2019 and spread across the whole country, affecting all 63 provinces and cities. In this study, ASF virus (ASFV) VN/Pig/HaNam/2019 (VN/Pig/HN/19) strain was isolated in primary porcine alveolar macrophage (PAM) cells from a sample originating from an outbreak farm in Vietnam’s Red River Delta region. The isolate was characterised using the haemadsorption (HAD) test, real-time PCR, and sequencing. The activity of antimicrobial feed products was evaluated via a contaminated ASFV feed assay. Phylogenetic analysis of the viral p72 and EP402R genes placed VN/Pig/HN/19 in genotype II and serogroup 8 and related it closely to Eastern European and Chinese strains. Infectious titres of the virus propagated in primary PAMs were 10⁶ HAD₅₀/ml. Our study reports the activity against ASFV VN/Pig/HN/19 strain of antimicrobial Sal CURB RM E Liquid, F2 Dry and K2 Liquid. Our feed assay findings suggest that the antimicrobial RM E Liquid has a strong effect against ASFV replication. These results suggest that among the Sal CURB products, the antimicrobial RM E Liquid may have the most potential as a mitigant feed additive for ASFV infection. Therefore, further studies on the use of antimicrobial Sal CURB RM E Liquid in vivo are required. Our study demonstrates the threat of ASFV and emphasises the need to control and eradicate it in Vietnam by multiple measures.
Show more [+] Less [-]Characterisation of fowl adenovirus (FAdV-8b) strain concerning the geographic analysis and pathological lesions associated with inclusion body hepatitis in broiler flocks in Turkey
2020
Cizmecigil, Utku Yusuf | Umar, Sajid | Yilmaz, Aysun | Bayraktar, Erhan | Turan, Nuri | Tali, Besim | Aydin, Ozge | Tali, Hasan Emre | Yaramanoglu, Mehmetcan | Yilmaz, Semaha Gul | Kolukisa, Ahmet | Sadeyen, Jean-Remy | Munir, Iqbal | Yılmaz, Hüseyin
Fowl adenovirus can cause important diseases in chickens such as inclusion body hepatitis, hepatitis hydropericardium syndrome, and gizzard erosion and ulceration. Inclusion body hepatitis has been regularly reported from many countries. This is the first case report from Turkey, describing an outbreak of inclusion body hepatitis in broiler farms due to fowl adenovirus-8b (FAdV-8b). Broiler flocks with mortality about 10% were visited in Turkey, and necropsy was performed on dead birds. Samples were subjected to PCR assay to detect FAdV and other viral pathogens. After sequencing, phylogenetic analysis was performed and the nucleotide sequences of hexon genes were compared with the FAdV sequences data available in GenBank. Clinical signs such as anorexia, depression, ruffled feathers, huddling, and greenish diarrhoea were observed. Mortality started at the 8ᵗʰ day of age and ranged from 10% to 14%. Necropsy showed severe hepatitis, jaundice, and pancreatitis. The main necropsy findings included a pale, enlarged, haemorrhagic, and friable liver along with swollen and haemorrhagic kidneys and spleen. PCR and sequence analysis revealed the presence of fowl adenovirus serotype 8b (FAdV-E). This is the first report on characterisation and the pathological lesions associated with FAdV in broilers in Turkey. Our findings suggest that FAdV strains could be an emerging pathogen in Turkish broilers and could actively contribute to hepatitis and immunosuppression.
Show more [+] Less [-]Meta-analysis of genetic diversity of the VP1 gene among the circulating O, A, and SAT2 serotypes and vaccine strains of FMD virus in Egypt
2020
El Nahas, Abeer F. | Salem, Sayed A.H.
Three strains of the FMD virus (A, O, and SAT 2) were recognised as causes of the FMD circulating in Egypt. The aims of this study were to trace the FMDV isolates from outbreaks in Egypt to understand their epidemiology and evolution and to understand the situation of the vaccine strains compared with the circulating serotypes. A meta-analysis was carried out by using the data available for FMD outbreaks in Egypt from GenBank and the World Reference Laboratory for Foot-and-Mouth Disease (WRLFMD); a comparison was done with both data sets for the three serotypes. MEGA-X was used for the evolution analysis, through constructions of phylogenetic trees for all sequences recorded in GenBank for each serotype in different Egyptian outbreaks in different years and also within the same year. Additionally, nucleotide substitution rate, molecular clock, and mean evolutionary rates were estimated for the three serotypes to understand and compare their evolution. Absence of some records of certain serotype outbreaks from the WRLFMD database was noted as were subsequent missing appropriate vaccine programmes. Genetic variation was recorded among the virus isolates within the same years and also the vaccine strain was associated with up to 26 amino acid substitutions. The evolution rate of the SAT2 strain was the highest of the circulating strains. SAT2 had high amino acid substitution per year at an important immunogenic site (130–170), serotype A had less, and serotype O the least. The need for different strategies for vaccine serotype selection is indicated.
Show more [+] Less [-]Characterisation of porcine enteropathogenic Escherichia coli isolated in northeastern India
2020
Kylla, Hosterson | Dutta, Tapan Kumar | Roychoudhury, Parimal | Subudhi, Prasant Kumar | Lalhruaipuii, | Lalsiamthara, Jonathan | Mandakini, Rajkumari
Enteropathogenic Escherichia coli (EPEC) is one of the main pathotypes causing gastroenteritis, particularly in young immunocompromised hosts. The study reports the prevalence, characterisation, and molecular epidemiology of EPEC from piglets in northeastern India. A total of 457 faecal samples were collected, from which 1,286 E. coli strains were isolated and screened by PCR. The resultant EPEC strains were serotyped and phenotypically characterised for resistance against 15 antimicrobials. Also, the phylogenetic sequence was analysed for 11 selected strains. A total of 42 strains (3.26%) belonged to atypical EPEC, of which, 15 (35.71%, and 2.29% of the 654 strains from this farm type) were isolated from organised and 27 (64.29%, and 4.27% of the 632 strains from this farm type) from unorganised farms; further, 5 (11.90% of the EPEC strains and 1.51% of the 330 strains from this breed) were isolated from the indigenous breeds and 37 (88.10%, and 3.87% of the 956 strains from this breed) from crossbred piglets. Serogroups O111 (11.9%) and O118 (7.14%) were the most prevalent of the 10 present. Sequence analysis of a length of the eaeA gene of 11 isolates of the region showed them to have 100% homology with each other and their identity ranged from 99.4% to 99.7% with GenBank reference sequences. All the EPEC isolates were multi-drug resistant, showing the highest resistance to amoxicillin (80.9%) and cephalexin (76.19%). The study highlighted the association of EPEC with piglet’s diarrhoea in northeastern India. EPEC isolates belonged to many serotypes and phenotypically all were multi-drug resistant with close genetic homology.
Show more [+] Less [-]Highly pathogenic avian influenza H5N8 in Poland in 2019–2020
2020
Śmietanka, Krzysztof | Świętoń, Edyta | Kozak, Edyta | Wyrostek, Krzysztof | Tarasiuk, Karolina | Tomczyk, Grzegorz | Konopka, Bogdan | Welz, Mirosław | Domańska-Blicharz, Katarzyna | Niemczuk, Krzysztof
Repeated incursions of highly pathogenic avian influenza virus (HPAIV) H5 subtype of Gs/GD lineage pose a serious threat to poultry worldwide. We provide a detailed analysis of the spatio-temporal spread and genetic characteristics of HPAIV Gs/GD H5N8 from the 2019/20 epidemic in Poland. Samples from poultry and free-living birds were tested by real-time RT-PCR. Whole genome sequences from 24 (out of 35) outbreaks were generated and genetic relatedness was established. The clinical status of birds and possible pathways of spread were analysed based on the information provided by veterinary inspections combined with the results of phylogenetic studies. Between 31 December 2019 and 31 March 2020, 35 outbreaks in commercial and backyard poultry holdings and 1 case in a wild bird were confirmed in nine provinces of Poland. Most of the outbreaks were detected in meat turkeys and ducks. All characterised viruses were closely related and belonged to a previously unrecognised genotype of HPAIV H5N8 clade 2.3.4.4b. Wild birds and human activity were identified as the major modes of HPAIV spread. The unprecedentedly late introduction of the HPAI virus urges for re-evaluation of current risk assessments. Continuous vigilance, strengthening biosecurity and intensifying surveillance in wild birds are needed to better manage the risk of HPAI occurrence in the future.
Show more [+] Less [-]Analysis of bovine tuberculosis transmission in Jalisco, Mexico through whole-genome sequencing
2020
Escárcega, Dulce Anahy Verdugo | Razo, Claudia Angélica Perea | Ruíz, Sara González | Gallegos, Susana Lucia Sosa | Suazo, Feliciano Milián | Alarcón, Germinal Jorge Cantó
Bovine tuberculosis, caused by M. bovis, is endemic in Mexico and has had a big impact on public health. Jalisco is considered to be an important dairy region in the country, accounting for approximately 19% of the total milk production. Within Jalisco, the region of Altos Sur holds the largest proportion of the cattle inventory of the state. To determine the frequency of bovine tuberculosis in Altos Sur, Jalisco, as well as M. bovis genetic diversity, sampling of tissue (lymph nodes, lungs, and liver) from Holstein cattle was performed in four abattoirs belonging to three municipalities of this region (Tepatitlán de Morelos, San Miguel el Alto, and Arandas). Spoligotyping and whole-genome sequencing were carried out to assess the genetic relationships of M. bovis strains circulating in this area, as well as a comparison to isolates from other places in Mexico. Prevalence was 15.06%, and distribution similar among the three municipalities. The most frequent spoligotypes were SB0673, SB121, and SB0145. Whole-genome sequencing revealed three main clades (I, II, III), but isolates did not show clustering by region. Phylogenetic analysis suggested ongoing transmission between herds of the different regions, and no unique source of infection was determined. This hinders efforts under the national program for the control and eradication of the disease, so serious attention must be paid to rural regions such as Altos Sur in order to improve its success.
Show more [+] Less [-]Malignant catarrhal fever in cattle in the Irkutsk Region
2020
Zakharova, Olga | Toropova, Nadezhda | Burova, Olga | Titov, Ilya | Meltsov, Ivan | Blokhin, Andrey
Malignant catarrhal fever (MCF) is a rare, under-explored lethal viral infection of cattle with gammaherpesvirus aetiological agents. Most often, the disease occurs on farms where cattle and sheep are kept together. However, other trigger mechanisms and environmental factors contribute. This study investigates the causation of MCF. An outbreak of MCF occurred in June - August 2017 in Kharchev village in Irkutsk Oblast, Russia. In this paper, we provide epidemiological (sanitary status of pastures, watering places, and premises) and weather data during the outbreak, and descriptions of the clinical signs and post-mortem changes in cattle. The virus was detected and isolated from pathological material samples and identified by molecular methods. Extreme weather conditions, mixed-herd cattle and sheep farming, and unsatisfactory feed quality contributed to the outbreak. A virus related to herpesvirus OvHV2 was isolated and typed (MCF/Irkutsk/2017). Phylogenetic analysis showed its close genetic relationship to isolates from cattle and sheep in Germany, USA, and the Netherlands. Sporadic outbreaks of MCF caused by biotic and abiotic factors together are typical for the Russian Federation, and the Irkutsk outbreak epitomised this. Temperature anomalies caused pasture depletion, resulting in feed and water deficiency for grazing animals and dehydration and acidosis. Heat stress in animals ultimately led to the occurrence of MCF in the herd.
Show more [+] Less [-]Molecular characterisation and genetic diversity of canine parvovirus type 2 prevalent in Central China
2020
Hu, Wen | Xu, Xin | Liu, Qiang | Ji, Jun | Kan, Yunchao | Yao, Lunguang | Bi, Yingzuo | Xie, Qingmei
Canine parvovirus (CPV) disease is one of the most threatening to domestic and wild dogs. A total of 132 clinical samples were isolated from domestic dogs with diarrhoea from Henan, Hubei, Jiangsu, and Anhui provinces from 2016 to 2017, and 56 were positive for CPV-2 by PCR. A phylogenetic tree was constructed for the isolate sequences incorporating 53 non-Chinese reference strains. VP2 sequences showed the strains mainly to be new CPV-2a/2b and CPV-2c genotypes. The Ala5Gly, Phe267Tyr, Ser297Ala, Tyr324Ile, Gln370Arg, Asn426Asp or Asn426Glu, and Thr440Ala sites in the VP2 protein antigenic region were found to have high mutation rates. The VP2 tertiary structural model shows that the change at these mutation points is a factor for the changes in the protein structure. Significant differences between the Central Chinese strains and others were found, indicating that evolution is geographically related and extended in major regions. The homology between the identified strains confirmed their relationship. Phylogenetic analysis indicated that the common genotypes in the same clusters differ slightly in homology and evolutionary history. This epidemiological study enriches the available data and serves as an important reference for studies on the evolution of CPV and selection of vaccines in China.
Show more [+] Less [-]Molecular study of Cysticercus tenuicollis from slaughtered sheep in Sulaymaniyah province, Iraq
2020
Mohammed, Aram Ahmad
Cysticercosis caused by the larval stage of Taenia hydatigena is economically the most important endemic parasitic disease in Iraq. Few data are available relating to the genetic divergence of this helminth. This study aimed to molecularly characterise Cysticercus tenuicollis isolates from sheep in Sulaymaniyah province, Iraq. DNA extraction and amplification of specimens of C. tenuicollis from 46 sheep were conducted by PCR for the mitochondrial 12S rRNA gene. The 19 amplicons were subjected to purification and partial sequencing. Five 12S rRNA nucleotide sequence haplotypes were found. The pairwise nucleotide difference between haplotypes of 12S rRNA gene ranged from 0.2% to 0.7%. Four out of the five haplotypes of C. tenuicollis contained one to two base mutations and were discovered in Iraq for the first time, and this may be a unique mutation globally which has not been recorded previously. Three newly recorded haplotypes contained only one single mutation, and the other one contained two mutations. Phylogenetic analysis showed that all isolated strains were closely related to Iranian sheep isolates. Four new strains of T. hydatigena were discovered for the first time in the study area.
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