Molecular Characterization of the env Gene of Bovine Leukemia Virus in Cattle from Pakistan with NGS-Based Evidence of Virus Heterogeneity

Characterization of the global genetic diversity of the bovine leukemia virus (BLV) is an ongoing international research effort. Up to now BLV sequences have been classified into eleven distinct genotypes. Although BLV genotyping and molecular analysis of field isolates were reported in many countries, there is no report describing BLV genotypes present in cattle from Pakistan. In this study we examined 27 env gene sequences from BLV-infected cattle coming from four farms located in Khyber Pakhtunkwa, Gilgit Baltisan and Punjab provinces. Phylogenetic analyses revealed the classification of Pakistani sequences into genotypes G1 and G6. The alignment with the FLK-BLV sequence revealed the presence of 45 mutations, namely, seven in genotype G1 and 33 in genotype G6. Five mutations were found in both, G1 and G6 genotypes. Twelve amino acid substitutions were found in the analyzed sequences, of which only one P264S was specific for sequences from Pakistan. Furthermore, a certain degree of nucleotide heterogeneity was identified by NGS. These results highlight the need for further study on the importance of genetic variability of BLV, especially in the context of its pathogenicity and potential effect on serological detection.

Morphological and pathogenic characterization of Fusarium oxysporum in lulo (Solanum spp.)

Fusarium oxysporum is one of the most limiting fungal pathogens of lulo crop. To determine its pathogenicity, this work morphologically and pathogenically characterized F. oxysporum isolates from different lulo-growing municipalities of the Department of Nariño. Twenty isolates were evaluated through a completely randomized design with two factors and three replicates per treatment, including a control. The first factor corresponded to 20 isolates of F. oxysporum and the second to 10 lulo genotypes. The morphological characterization involved determining growth rate (GR), color (CO), mycelial type (MT), medium coloration (Mc), shape (Sh), size (S), number (N) of macroconidial (Ma) and microconidial (Mi) septa, presence of chlamydospores (PC), and chlamydospore shape (CS). Moreover, the pathogenic characterization was based on the incubation period (IP), absolute growth rate (AGR), disease severity (DS), disease incidence (I), and vascular discoloration (VD). The morphological characterization demonstrated that all isolates corresponded to F. oxysporum. For IP, genotype G1 showed the lowest average at 18 days. For AGR, genotype G2 had the lowest height increase at 0.05 cm.day-1. For DS, genotype G1 reached the highest severity level (level 9) and a disease incidence of 100%. This study provides the first report of the special form of F. oxysporum f. sp. quitoense in Nariño. Solanum hirtum, Solanum sessiliflorum, and Solanum estramonifolium were resistant to the isolates evaluated, demonstrating that wild species should be considered as sources of resistance for breeding programs aiming to obtain resistant commercial genotypes.

Efficacy of novel albendazole salt formulations against secondary cystic echinococcosis in experimentally infected mice

In this study, we evaluated the efficacy, expressed as a mean weight decrease of the whole echinococcal cyst mass, of novel benzimidazole salt formulations in a murine Echinococcus granulosus infection model. BALB/c mice were intraperitoneally infected with protoscoleces of E. granulosus (genotype G1). At 9 months post-infection, treatment with albendazole (ABZ), ricobendazole (RBZ) salt formulations, and RBZ enantiomer salts (R)-(+)-RBZ-Na and (S)-(−)-RBZ-Na formulations were initiated. Drugs were orally applied by gavage at 10 mg kg−1 body weight per day during 30 days. Experimental treatments with benzimidazole sodium salts resulted in a significant reduction of the weight of cysts compared to conventional ABZ treatment, except for the (S)-(−)-RBZ-Na enantiomer formulation. Scanning electron microscopy and histological inspection revealed that treatments impacted not only the structural integrity of the parasite tissue in the germinal layer, but also induced alterations in the laminated layer. Overall, these results demonstrate the improved efficacy of benzimidazole salt formulations compared to conventional ABZ treatment in experimental murine cystic echinococcosis.

Genotyping of Fresh and Parafinized Human Hydatid Cysts Using nad1 and cox1 Genes in Hamadan Province, West of Iran

Background: Hydatidosis is a cosmopolitan zoonotic infection and Hamadan Province in the west of Iran is one of the most important foci of human hydatidosis in Iran. The aim of the current study was the genetic characterization of hydatid cysts operated from humans in Hamadan Province. atid cysts operated from humans in Hamadan Province. Methods: Seventy-two hydatid cysts samples including 50 paraffinized and 22 fresh human hydatid cysts collected from different hospitals in Hamadan Province, western Iran. The cysts' DNA genome was extracted by kit and PCR was performed for amplifying the fragments of 400 and 450bp for nad1 and cox1 mitochondrial genes, respectively. Genotype diversity and sequence variations of the cysts' isolates were studied by related software. Results: DNA from all (100%) paraffinized and fresh hydatid cysts samples extracted successfully. All paraffinized and fresh hydatid cysts samples were amplified by PCR assay using nad1gene, however, only 18 and 8 samples from paraffinized and fresh hydatid cyst samples was amplified using cox1 gene, respectively. The sequences analysis indicated that, 98.61% the Echinococcus granulosus samples were belong to the genotype G1 and 1.39% were G3 genotype. Conclusion: Genotypes of E. granulosus in human samples in Hamadan Province are G1 and G3 and these findings are proved by phylogenic analysis.

Parasitological Contamination of Vegetables Sold in Tunisian Retail Markets with Helminth Eggs and Protozoan Cysts

ABSTRACT Foodborne diseases continue to represent an important threat to public health in many parts of the world and are particularly widespread in developing countries. They are essentially acquired through an oro-fecal route via the consumption of uncooked fruits and vegetables. This study evaluated the parasitological contamination of vegetables for sale to humans in Tunisian retail markets. A total of 240 samples of fresh vegetables were examined for helminth eggs and protozoan cysts and oocysts (collectively, (oo)cysts) contamination. The parasitic elements (helminth eggs and protozoan (oo)cysts) were concentrated by sucrose flotation and identified by microscopic examination. The molecular identification of Echinococcus granulosus eggs was carried out using PCR. Helminth eggs and protozoan (oo)cysts eggs were found in 12.5% of the unwashed vegetables, and the most common parasites observed in vegetables were coccidian oocysts (4.1%), Toxocara spp. (2.5%), hookworm (2.1%), and Taenia spp. (1.25%) eggs, followed by Pseudolimax butschlii (1.6%) and Entamoeba coli (1.6%) protozoan cysts. Furthermore, parasite contamination differed significantly from one city to another. Taeniid eggs were identified by PCR as E. granulosus sensu stricto (s.s.) (genotype G1). To our knowledge, this study highlights for the first time in Tunisia that fresh vegetables for sale in markets are contaminated with helminths and protozoan cysts, which are potentially pathogenic for humans. The control of these pathogens is in part a question of sanitary education, especially for retail vendors, and in part of improvement in hygiene measures throughout the food production chain, from the field to the consumer. HIGHLIGHTS

Draft Genome Sequence of Avian Chlamydia abortus Genotype G1 Strain 15-70d24, Isolated from Eurasian Teal in Poland

Here, we report the draft genome sequence of avian Chlamydia abortus genotype G1 strain 15-70d24, isolated from Eurasian teal in Poland. The total genome assembly length is 1,149,382 bp.

A systematic review and meta-analysis of the genetic characterization of human echinococcosis in Iran, an endemic country

Human echinococcosis is an infectious disease caused by tapeworms belonging to the species Echinococcus. This parasite has a worldwide distribution and is considered a neglected tropical disease by the World Health Organization. Due to the diversity of Echinococcus spp. hosts, as well as variation in geographical, climatic, and socio-ethnic conditions, the question of the strains or genotypes of Echinococcus spp. that are involved in human infections is important. The aim of this study was to provide a summary of the available data on genotypes of Echinococcus obtained from the Iranian population. Four international databases (PubMed, Scopus, Science Direct, and Web of Science) and 4 Persian databases (Magiran, Scientific Information Database, Iran Medex, and IranDoc) were searched for cross-sectional studies that reported the genotypes of Echinococcus spp. in human echinococcosis cases using molecular methods in Iran through July 2018. The Newcastle-Ottawa Scale was used to assess the quality of the selected studies. A total of 559 cases of human cystic echinococcosis were reported in the 21 included articles. The majority of cases belonged to genotype G1 (89.2%; 95% confidence interval [CI], 80.1 to 95.8), genotype G6 (8.2%; 95% CI, 2.8 to 15.9), and genotype G3 (2.3%; 95% CI, 1.1 to 3.9). Since genotype G1 of Echinococcus appears to be the most prevalent genotype affecting humans in Iran, disease control initiatives aimed at sheep intermediate hosts may be the most beneficial. In addition, educational programs and serological screening in individuals may help reduce the national impact of the disease.