Helminth infection and human mobility in sambaquis: Paleoparasitological, paleogenetic, and microremains investigations in Jabuticabeira II, Brazil (2890 ± 55 to 1805 ± 65 BP)

Sambaquis or shellmounds are archeological sites constructed by hunter-fisher-gatherers that inhabited the Brazilian coast about 10,000–2000 yrs BP. Jabuticabeira II (JABII: 2890 ± 55 to 1805 ± 65 BP) is one of dozens of contemporaneous sambaquis of the Santa Catarina state, South Brazil, and contains hundreds of neatly organized burials, indicating great population density. In order to gather information about the health, diet and way of life of people in JABII, a paleoparasitological, paleogenetic, and micro-human remains investigation was carried out. Pelvic region and environmental control samples from six individuals exhumed from JABII were submitted to microscopic and ancient DNA (aDNA) investigation. Paleoparasitological analyses based on light microscopy were negative. However, a variety of informative microremains were found. Diatoms, fish scales, and algae characterize the marine and estuarine environment. Ipomoea batatas and Zea mays starch grains suggested cultivated items as part of their diet in agreement with the literature. The finding of Podocarpus sp. pollen grain, characteristic of highlander vegetation, suggests human mobility of JABII individuals which were settlement in the coast. Paleogenetic analyses showed Ascaris sp. helminth infection based on nad1 gene fragment detected from an individual excavated at L3 FS7 burial (1826 ± 40 BP). This aDNA result places the antiquity of Ascaris sp. infection, and haplotypes that are circulating in humans and other animals nowadays, in Pre-Columbian South American times.

Prevalence and Molecular Characterization of Toxocara cati Infection in Feral Cats in Alexandria City, Northern Egypt

Background: This study was performed to determine the prevalence and to identify precisely Toxocara spp., which infects feral cats in Alexandria, Egypt based on morphological and molecular approaches. Methods: This cross-sectional study was carried out on 100 feral cats trapped from different areas of Alexandria during 2018. Adult male and female worms were recovered from small intestinal contents after euthanasia and dissection of cats. Distinct morphological features were initially determined using available keys, and then after amplification and sequencing of the mitochondrial NADH dehydrogenase subunit 1 (nad1) gene was carried out and phylogenetic trees were constructed. Results: Forty out of 100 cats were infected with Toxocara spp. Intensity of infection ranged from 1 to 9 worms/cat, with a mean of 2.27±1.6. All isolates were confirmed as T. cati based on morphological features and the sequence of nad1 gene. Results of the current study clearly show that Egyptian T. cati isolate examined herein is genetically similar to those recorded in other countries. Conclusion: The current work revealed high prevalence of T. cati in feral cats in the study area. This is the first genetic study that confirms T. cati from feral cats in Egypt. In addition, it demonstrated the suitability and need of genetic markers such as nad1 for identification of Toxocara spp. Furthermore highlights the public health importance of T. cati in Egypt.

Employing NADH Dehydrogenase Subunit 1 in the Determination of Echinococcus granulosus Strain in Sheep, Cattle and Human in Thi-Qar Province, Iraq

Echinococcosis is a zoonotic disease caused by the larval stage of the tapeworm Echinococcus granulosus. This disease is an important public health and a significant economic issue in Iraq, where the lungs and livers are the popular places of infection. The aim of the current study focused on using the molecular techniques in the detection of an E. granulosus strain that causes cystic echinococcosis to human, sheep and cattle in Thi-Qar province, Iraq. In the current study, thirty isolates of E. granulosus were collected from 10 human hydatid cysts through surgery done at Al-Hussein Imam Teaching Hospital in Thi-Qar province and 10 sheep with 10 cattle hydatid cysts were obtained from the slaughterhouse in Thi-Qar province, Southern of Iraq to identify strains of E. granulosus which infect human and other intermediate hosts (sheep and cattle). The molecular study was carried out on the isolates and a specific primer set for the mitochondrial dehydrogenase NADH subunit 1 (NAD1) gene was used. This primer set was amplified 400 bp of the NAD1 gene in all selected isolates. The PCR products for the twelve selected isolates of E. granulosus (4 isolates per intermediate host) were sequenced and the results for these twelve isolates showed that all sequenced isolates, except one isolate Eg_5, belonged to the sheep strain G1 and a slight genetic diversity was observed with the reference sequences of the strain G1. The exception was in the isolate Eg_5 isolated from a cattle liver, which was similar to the buffalo strain G3. This study concludes that the common E. granulosus strain in Thi-Qar province is G1.

Emerging risk of Dirofilaria spp. infection in Northeastern Europe: high prevalence of Dirofilaria repens in sled dog kennels from the Baltic countries

Dirofilariasis is a fast-spreading disease of dogs and humans in Europe. We investigated whether Dirofilaria spp. have spread northwards in Europe, invading the Baltic countries. Altogether, 424 blood samples were collected from eight countries in the period 2017–2019, including 227 samples from sled dogs and 197 samples from other dogs. PCR amplification and sequencing were conducted employing three genetic markers (mitochondrial [mt] 12S rDNA, mt cytochrome c oxidase subunit I [COI] gene and mt dehydrogenase subunit I [NAD1] gene). The SNAP test (IDEXX) for detection of D. immitis infections was also implemented. The DNA of D. repens was detected in 59 of 424 dogs (prevalence 13.9%). D. repens was found in sled dogs from Lithuania, Latvia, Poland and Belarus. Only one dog from Estonia was infected, apparently an imported case. The highest prevalence was recorded in Lithuania (38%). Among pet dogs from the Ukraine, six dogs tested positive (3.8%). Our study has revealed a high prevalence of D. repens infections in Lithuania and Latvia, but no evidence for spread of the heartworm D. immitis. We conclude that sled dog kennels constitute hot spots for D. repens transmission.

Sequencing and phylogenetic analysis of mitochondrial cox1 and nad1 genes in Toxocara canis and Toxascaris leonina isolates from Iran

Toxocara canis and Toxascaris leonina are the most important ascaridoid nematodes of the family Toxocaridae. The present study was aimed to characterisation and analysis of genetic variation within and among T. canis and T. leonina isolates obtained from Iran by sequencing partial mitochondrial cytochrome c oxidase subunit 1 (pcox1) and partial NADH dehydrogenase subunit 1 (pnad1) genes. A total number of 134 adult nematodes belonging to Toxocaridae family were collected from stray dogs in Alborz province, Iran during 2015 and 2016. Polymerase chain reaction (PCR) was performed and products were sequenced. Sequences of two mitochondrial cox1 and nad1 genes were compared with other sequences in the GenBank, while multiple sequences alignment analysis was performed using the Bioedit and MEGA6 software and phylogenetic tree was plotted. For all isolates, amplicons of about 450 and 350 base pairs (bp) were successfully produced by PCR for cox1 and nad1, respectively. All sequences of T. canis isolates from present study were 100% homologous across the nad1 gene but not in the cox1 gene. The results indicate that the PCR method based on sequence of cox1 and nad1 genes is a suitable technique for the differentiation of T. canis and T. leonina species and that mtDNA regions could be used as genetic markers for the identification and differentiation of Toxocara species.

Two Human Cases of Echinococcus ortleppi Infection in the Lung and Heart in Vietnam

This is a report of 2 cases of human hydatidosis caused by Echinococcus ortleppi in Vietnam. The patients were a 12-year-old male (case 1) having a cyst of 10.0×9.0 cm size in the lung and a 50-year-old female with a 3.0×3.3 cm-sized cyst in the heart. Eosinophilia was 33.7% in the male and 45.8% in the female patient. C-reactive protein was increased to 16.5 mg/L in the male and 18.2 mg/L in the female. Both patients were positive for ELISA at OD=2.5 and 3.1, respectively. Echinococcus protoscolices were collected from the cysts by amniocentesis and surgery. The protoscolices were identified as E. ortleppi by morphology and analysis of mitochondrial NADH dehydrogenase 1 (nad1) gene sequence. Both patients were cured by surgical resection of the hydatid cyst combined with albendazole medication. The E. ortleppi infection in lung is the second report, and the other in the heart is the first in Vietnam.

Fasciola hepatica in Brazil: genetic diversity provides insights into its origin and geographic dispersion

Fasciola hepatica is a trematode parasite that affects mammals, including humans. In Brazil, fascioliasis, a disease caused by the parasite, is of great importance. The disorder affects the welfare of the Brazilian population through impairing the agricultural production of cattle, where the disease causes weight loss as a result of liver damage. This study aimed to evaluate the genetic diversity of F. hepatica throughout Southern Brazil to determine its geographic origin and estimate the colonization route of the parasite. To accomplish these aims, flukes were collected from slaughterhouses in three endemic areas of Rio Grande do Sul and Paraná states. DNA was isolated using the phenol–chloroform protocol from single flukes and two mitochondrial genes, cytochrome oxidase subunit I (COI) and nicotinamide dehydrogenase subunit 1 (Nad1), were amplified and sequenced. Ten haplotypes of COI were found from 75 isolated parasites and the total haplotype and nucleotide diversity observed were 0.475 and 0.002, respectively. Using the Nad1 gene, we found 24 haplotypes from 79 samples, resulting in haplotype and nucleotide diversity values of 0.756 and 0.004, respectively. An analysis of molecular variance showed that 57.4% and 77.5% of variation was within populations (FST), while 9.0 and 36.8% of variation was among groups (FCT) when considering COI and Nad1 genes, respectively. For COI, the fixation index values of 0.425 and 0.368 were obtained for FST and FCT, respectively, while analysis of Nad1 0.225 and 0.089 index values were obtained for FST and FCT, respectively. We have determined that F. hepatica found in the two distinct areas originated from several geographical regions, since we found haplotypes that were shared with at least three different continents. These data are in accordance with the recent colonization of Brazil, and the recent import of cattle from South American, European and, possibly, some African countries. The observed FST and FCT values for COI and Nad1 genes of F. hepatica may be a result of limited movement of animals within states and support the lack of geographical structure of the parasite in Brazil, which are in agreement with the observed cattle production systems in this region.