Molecular Phylogenetics of Physa acuta (Pulmonata: Basommatophora): an Invasive species in Central Punjab Pakistan

Physids belong to Class Gastropoda; belong to Phylum Mollusca and being bioindicators, intermediate hosts of parasites and pests hold a key position in the ecosystem. There are three species of Genus Physa i.e. P. fontinalis, Physa acuta and P. gyrina water bodies of Central Punjab and were characterized on the basis of molecular markers High level of genetic diversity was revealed by polymorphic RAPD, however SSR markers were not amplified. The multivariate analysis revealed polymorphism ranging from 9.09 percent to 50 percent among the three Physid species. Total number of 79 loci were observed for the three species under study and 24 loci were observed to be polymorphic. These RAPD fragment(s) can be developed into co dominant markers (SCAR) by cloning and can be further sequenced for the development of the Physa species specific markers to identify the introduced and native species in Pakistan.

Diseases of Gastropoda

Gastropods (class Gastropoda) form the largest of the classes in the phylum Mollusca and inhabit terrestrial, fresh water and marine environments. A large number of these species are of major conservation importance and are an essential component of ecosystems. Gastropods may be deemed as pests, having a negative impact in horticulture and agriculture, whereas others may be used as a food source for human consumption and therefore are beneficial. Gastropods are susceptible to primary diseases and also act as intermediate hosts for diseases which affect other animals, including humans. The diseases described include two that are notifiable to the World Organisation for Animal Health (OIE): Xenohaliotis californiensis and Abalone viral ganglioneuritis caused by Haliotid herpesvirus-1 (HaHV-1). Research into the diseases of gastropods has often focused on those species that act as intermediate disease hosts, those that are used in research or those cultured for food. In this paper we review the viral, bacterial, fungal, parasitic and miscellaneous conditions that have been reported in gastropods and mention some of the factors that appear to predispose them to disease. The pathogenicity of a number of these conditions has not been fully ascertained and more research is needed into specifying both the etiological agent and significance in some of the diseases reported.

Infection of the Asian gray shrew Crocidura attenuata (Insectivora: Soricidae) with Sarcocystis attenuati n. sp. (Apicomplexa: Sarcocystidae) in China

Background Data on the genus Sarcocystis in insectivores are limited. The Asian gray shrew Crocidura attenuata is one of the most common species of the insectivore family Soricidae in South Asia and Southeast Asia. To our knowledge, species of Sarcocystis have never been recorded previously in this host. Methods Tissues were obtained from 42 Asian gray shrews caught in 2017 and 2018 in China. Sarcocysts were observed using light microscopy (LM) and transmission electron microscopy (TEM). To describe the parasite life cycle, muscle tissues of the host infected with sarcocysts were force-fed to two beauty rat snakes Elaphe taeniura. Individual sarcocysts from different Asian gray shrews, and oocysts/sporocysts isolated from the small intestines and feces of the experimental snakes, were selected for DNA extraction, and seven genetic markers, namely, two nuclear loci [18S ribosomal DNA (18S rDNA) and internal transcribed spacer region 1 (ITS1)], three mitochondrial genes [cytochrome oxidase subunit 1 (cox1), cox3 and cytochrome b], and two apicoplast genes (RNA polymerase beta subunit and caseinolytic protease C), were amplified, sequenced and analyzed. Results Sarcocysts were found in 17 of the 42 (40.5%) Asian gray shrews. Under LM, the microscopic sarcocysts showed saw- or tooth-like protrusions measuring 3.3–4.5 μm. Ultrastructurally, the sarcocyst wall contained numerous lancet- or leaf-like villous protrusions, similar to those described for type 9h of the common cyst wall classification. The experimental beauty rat snakes shed oocysts/sporocysts measuring 11.9–16.7 × 9.2–10.6 μm with a prepatent period of 10–11 days. Comparison of the newly obtained sequences with those previously deposited in GenBank revealed that those of 18S rDNA and cox1 were most similar to those of Sarcocystis scandentiborneensis recorded in the tree shrews Tupaia minor and Tupaiatana (i.e., 97.6–98.3% and 100% identity, respectively). Phylogenetic analysis based on 18S rDNA or ITS1 sequences placed this parasite close to Sarcocystis spp. that utilize small animals as intermediate hosts and snakes as the known or presumed definitive host. On the basis of morphological and molecular characteristics and host specificity, the parasite was proposed as a new species, named Sarcocystis attenuati. Conclusions Sarcocysts were recorded in Asian gray shrews, to our knowledge for the first time. Based on morphological and molecular characterization, a new species of parasite is proposed: Sarcocystisattenuati. According to the LM and TEM results, S. attenuati sarcocysts are distinct from those of Sarcocystis spp. in other insectivores and those of S. scandentiborneensis in tree shrews. The 18S rDNA or cox1 sequences of Sarcocystis attenuati shared high similarity with those of Sarcocystisscandentiborneensis, Sarcocystis zuoi, Sarcocystis cf. zuoi in the Malayan field rat, and Sarcocystis sp. in the greater white-toothed shrew. Therefore, we suggest that more research on the relationships of these closely related taxa should be undertaken in the future. Graphical abstract

Middle East respiratory syndrome coronavirus infection in camelids

Middle East respiratory syndrome coronavirus (MERS-CoV) is the cause of a severe respiratory disease with a high case fatality rate in humans. Since its emergence in mid-2012, 2578 laboratory-confirmed cases in 27 countries have been reported by the World Health Organization, leading to 888 known deaths due to the disease and related complications. Dromedary camels are considered the major reservoir host for this virus leading to zoonotic infection in humans. Dromedary camels, llamas, and alpacas are susceptible to MERS-CoV, developing a mild-to-moderate upper respiratory tract infection characterized by epithelial hyperplasia as well as infiltration of neutrophils, lymphocytes, and some macrophages within epithelium, lamina propria, in association with abundant viral antigen. The very mild lesions in the lower respiratory tract of these camelids correlate with absence of overt illness following MERS-CoV infection. Unfortunately, there is no approved antiviral treatment or vaccine for MERS-CoV infection in humans. Thus, there is an urgent need to develop intervention strategies in camelids, such as vaccination, to minimize virus spillover to humans. Therefore, the development of camelid models of MERS-CoV infection is key not only to assess vaccine prototypes but also to understand the biologic mechanisms by which the infection can be naturally controlled in these reservoir species. This review summarizes information on virus-induced pathological changes, pathogenesis, viral epidemiology, and control strategies in camelids, as the intermediate hosts and primary source of MERS-CoV infection in humans.

Another plea for ‘best practice’ in molecular approaches to trematode systematics: Diplostomum sp. clade Q identified as Diplostomum baeri Dubois, 1937 in Europe

DNA sequence data became an integral part of species characterization and identification. Still, specimens associated with a particular DNA sequence must be identified by the use of traditional morphology-based analysis and correct linking of sequence and identification must be ensured. Only a small part of DNA sequences of the genus Diplostomum (Diplostomidae) is based on adult isolates which are essential for accurate identification. In this study, we provide species identification with an aid of morphological and molecular (cox1, ITS-5.8S-ITS2 and 28S) characterization of adults of Diplostomum baeri Dubois, 1937 from naturally infected Larus canus Linnaeus in Karelia, Russia. Furthermore, we reveal that the DNA sequences of our isolates of D. baeri are identical with those of the lineage Diplostomum sp. clade Q , while other sequences labelled as the ‘D. baeri’ complex do not represent lineages of D. baeri. Our new material of cercariae from Radix balthica (Linnaeus) in Ireland is also linked to Diplostomum sp. clade Q. We reveal that D. baeri is widely distributed in Europe; as first intermediate hosts lymnaeid snails (Radix auricularia (Linnaeus), R. balthica) are used; metacercariae occur in eye lens of cyprinid fishes. In light of the convoluted taxonomy of D. baeri and other Diplostomum spp., we extend the recommendations of Blasco-Costa et al. (2016, Systematic Parasitology 93, 295–306) for the ‘best practice’ in molecular approaches to trematode systematics. The current study is another step in elucidating the species spectrum of Diplostomum based on integrative taxonomy with well-described morphology of adults linked to sequences.

Occurrence and genetic characterization of Echinococcus granulosus sensu lato from domestic animals in Central Iran

Background The species complex of Echinococcus granulosus sensu lato (s.l.) causes cystic echinococcosis distributed worldwide. There is no genotype information from hydatid cysts in the intermediate hosts in Central Iran. Therefore, in this study, we analyzed the hydatid cysts in livestock slaughtered in an abattoir in this region. Six hundred fifty-seven hydatid cysts were isolated from 97 animals, including sheep, cattle, camels, and goats slaughtered in Yazd abattoir from September 2018 to January 2020. The demographic data was collected as well as cyst location, fertility, and viability. Out of 657 samples, 164 samples were genotyped. Then, phylogenetic analysis was performed using MEGAX. Statistical analyses were done using SPSS version 16.0 by chi-square with a significant difference of less than 0.05. Results Out of 164 samples, the G1-G3 complex genotype had the most frequency in samples, with 135 cases recognized. The G6/G7 was observed in 19 isolates and G5 was reported in nine samples. One sample was detected as Taenia hydatigena. Conclusions This study showed that G1-G3 and G6/G7 genotypes were presented in all animals, but G5 was reported only in cattle, goats, and camels. It is the first molecular identification of cystic echinococcosis in Central Iran. Hence, reporting G5 in livestock in this area should be considered due to transmission to humans.

Temporal effects and changes in the parasitic community of Prochilodus lineatus (Valenciennes, 1837) (Characiformes: Prochilodontidae) in a floodplain

The construction of dams causes several impacts on aquatic environments, altering the flow of rivers, environmental variables, and all biota present, including parasites. Little is known about how the parasitic community can be influenced in the long term by environmental changes. In this study, it was expected that the impacts caused by environmental disturbances will be directly reflected by the composition of the parasite populations. We evaluated the change in the structure of the Prochilodus lineatus endoparasite community between two periods sampled 15 years apart in the upper Paraná River floodplain. There was a significant difference in the weight–length relationship of P. lineatus between these periods and a total of 15 species of parasites were found: 11 species in Period 1 and nine species in Period 2 and five species occurred in both periods. The species richness and diversity were higher in Period 1, and we observed that the correlation of descriptors (richness, diversity and evenness) increased with fish length in this period. In both periods, digeneans numerically dominated the parasitic community, and we verified changes in the composition of parasites between periods. Both the host and the parasites were possibly affected by the environmental impacts resulting from the construction of dams over time, and it is noteworthy that complex life cycle parasites such as Digenea and Acanthocephala require intermediate hosts to complete their life cycle, and the population responds to fluctuations in the face of modified environments.

An Emerging, Neglected and Underestimated Zoonotic Parasitic Ocular Infestation: A Comprehensive Review on Thelaziasis

Human thelaziasis is an emerging insect-borne zoonotic ocular parasitic infestation, occur more commonly in rural communities with poor living and low socioeconomic living, and mainly affects the children and old age people, where humans live in close proximity with animals. Human thelaziasis is caused by both Thelazia callipaeda and Thelazia californiensis. T. callipaeda lives under the eye lids, nictitating membranes, orbit, conjunctival sac, lachrymal glands, and lacrimal ducts of cats, dogs, rabbits, horses, cattle, deer, badgers, monkeys, wolves, foxes (definitive hosts) and man being an accidental host. The vectors (intermediate hosts) are non-biting, tear-seeking, diptera flies of family Drosophilidae (fruit flies) Phortica variegata, which feeds on tears of their definitive hosts, including humans. Clinical manifestations include conjunctivitis, lacrimation, itching or pain with foreign body sensation, epiphora, follicular hypertrophy, and less often with severe signs and symptoms such as keratitis, photophobia, ectropion, corneal opacities (due to the migration of worm across the cornea), floaters within the eye chamber leading to visual impairment/blindness. The knowledge and scientific information on human thelaziasis is still unknown or relatively limited to many ophthalmologists and clinicians, and received little attention; hence this comprehensive and systematic review of human thelaziasis, is undertaken to highlight its importance and further research.

Risk of Human Exposure to the Intestinal Schistosome, Schistosoma mansoni, across Seasons along the Senegal River

Background: Schistosomiasis is an emerging disease associated with changes to the environment that have increased human contact rates with disease-causing parasites, flatworms that are released from freshwater snails. For example, schistosomiasis remains a major public health problem in Northern Senegal, where prevalence in schoolchildren often reaches 90%. Aim: This study focuses on the impact of seasonality on the risk of human exposure (RHE) to Schistosoma mansoni, defined as the total number of cercariae (the free-living life stage that infects humans) shed from all Biomphalaria pfeifferi snails collected at a site using standardized methods. We focus on RHE because it is rarely quantified and a recent study demonstrated that snails stop shedding cercariae when snail densities increase and thus per capita snail resources become limited [2], suggesting that densities of snails might not be directly proportional to RHE to schistosomes. Method: We sampled four water access points in three villages every other week during the early (Dry1) and later dry seasons (Dry2) and the rainy season, quantifying the abundance of infected and non-infected snail intermediate hosts, cercariae released per infected snail, and water chemistry. We used simple and multiple linear regressions to assess how seasonality and environmental parameters affect non-infected and infected snail abundance and RHE. Results: Although RHE was found across all seasons, the abundance of infected and non-infected snail intermediate hosts and cercariae, as well as prevalence (23.71%), were all highest in the rainy season. In the rainy season, RHE was positively associated with the density of snail hosts and their periphyton food resource. Conclusion: Although previous studies have examined the influence of seasonality on snail densities, few studies have explored the effects of seasonality on cercarial densities, which is the primary source of infection to humans. Our study demonstrates that cercarial densities are greater in the rainy season than in the early or late dry seasons. Given that cercarial densities directly pose risk of infection to humans, unlike non-infected or infected snails, these finding should help to inform decision making and schistosomiasis control efforts in West Africa.

Pilot Malacology Surveys for the Intermediate Hosts of Schistosomiasis in Rural and Semi-Urban Areas of the Moyen-Ogooué Province, Gabon

The objective of this pilot malacological survey was to identify the snail intermediate hosts for Schistosoma haematobium in endemic rural and semi-urban areas of Gabon. Snails were collected, morphologically identified, and tested for infection by cercarial shedding. Released cercariae were morphologically identified using low-power light microscopy. A total of six species of snails were collected throughout the study area, with Bulinus truncatus, B. forskalii, and Potadoma spp. being the most predominant species collected. Only the Bulinus species were tested for infection by cercarial shedding, of which only B. truncatus shed cercariae. Some B. truncatus shed mammalian schistosome cercariae, while others shed Gymnocephalus cercariae. Our results indicate that B. truncatus appears to be a potential intermediate host of schistosomiasis in Gabon, where cases of S. haematobium, S. guineensis, and S. intercalatum infection are reported. However, it will be important to further understand the species diversity and transmission dynamics of schistosomes.