Analysis Of Multilocus Genotypes Research Articles

Molecular Genotyping Analysis Suggested High Clonality and Simple Marker Loci in Puccinia striiformis f. sp. tritici Population in Uzbekistan, Central Asian Region.

Puccinia striiformis f. sp. tritici (Pst) is a destructive pathogen that causes wheat stripe rust worldwide. Understanding the population structure and dynamic of pathogen spread is critical to fight against this disease. Limited information is available for the population genetic structure of Pst in Uzbekistan, Central Asia. In this study, we carried out surveillance from nine different regions (Andijan, Fergana, Jizzakh, Kashkadarya, Namangan, Samarkand, Sirdaryo, Surkhandarya, and Tashkent) of Uzbekistan to fill this gap. A total of 255 isolates were collected, which were genotyped using 17 polymorphic simple sequence repeat markers. The DAPC analysis results showed no population subdivision in these sample-collected regions except Surkhandarya. Multilocus genotype (MLG) analysis, FST, and Nei's genetic distance results indicated a clonal population (rBarD ≤ 0.12) and merely three MLGs accounting for 70% of the overall population. MLG-34 was predominant in all regions of Uzbekistan, followed by MLG-36 and MLG-42. Low genotypic diversity was observed in Andijan, Fergana, Jizzakh, Kashkadarya, Namangan, Sirdaryo, and Tashkent (0.56 to 0.76), compared with Samarkand (0.82) and Surkhandarya (0.97). No virulence against Yr5, Yr15, YrSp, and Yr26 was found, while resistance was overcome against Yr1, Yr2, Yr6, Yr9, Yr17, and Yr44 genes (virulence frequency ≥75%). Comparative study results of Uzbekistan with the previous Himalayan population showed divergence from China and Pakistan populations. Further studies need to be conducted in a worldwide context to understand migration patterns; for that purpose, collaborative work is essential because of the Pst long-distance migration capability.

Genetic Diversity Analysis of Anisogramma anomala in the Pacific Northwest and New Jersey.

Anisogramma anomala, a biotrophic ascomycete, causes eastern filbert blight (EFB) of hazelnuts (Corylus spp.). EFB is endemic in eastern North America, preventing the commercial production of European hazelnut (C. avellana L.). In contrast, the historic absence of A. anomala in the Pacific Northwest (PNW) supported the development of a robust hazelnut industry. Circa 1960, A. anomala was inadvertently introduced into southwestern Washington, causing orchard devastation. Distribution of the pathogen in the PNW has been hypothesized to be the result of a single-point introduction. This study aimed to investigate the single-point introduction hypothesis of A. anomala by comparing the genetic diversity of A. anomala samples from the PNW and New Jersey (NJ). Specimens from the main PNW production region (n = 60) and an area within the pathogen's native range, NJ (n = 151), were genotyped using 15 simple sequence repeat (SSR) markers. The following were used to assess genetic diversity and population structure: allelic summary statistics, discriminant analysis of principal components, network median-joining tree, analysis of multilocus genotypes, and allelic population diversity analysis. Analyses separated the samples into one cluster containing all the PNW isolates, and five clusters of NJ isolates. The PNW samples were nearly genetically uniform, and the NJ isolates were diverse. These findings support the hypothesis that A. anomala in the PNW was derived from a single-point introduction and corroborate previous studies that have shown A. anomala is very diverse in NJ. This indicates that maintaining restrictions on the movement of Corylus into the PNW is important to prevent the introduction of new populations of A. anomala, thus protecting the PNW hazelnut industry.

First report of Giardia duodenalis in dairy cattle and beef cattle in Shanxi, China.

Giardia duodenalis is an important intestinal parasitic protozoan that infects several vertebrates, including humans. Cattle are considered the major source of giardiasis outbreak in humans. This study aimed to investigate the prevalence and multilocus genotype (MLG) of G. duodenalis in Shanxi, and lay the foundation for the prevention and control of Giardiosis. DNA extraction, nested polymerase chain reaction, sequence analysis, MLG analysis, and statistical analysis were performed using 858 bovine fecal samples from Shanxi based on three gene loci: β-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi). The overall prevalence of G. duodenalis was 28.3%, while its prevalence in Yingxian and Lingqiu was 28.1% and 28.5%, respectively. The overall prevalence of G. duodenalis in dairy cattle and beef cattle was 28.0% and 28.5%, respectively. G. duodenalis infection was detected in all age groups evaluated in this study. The overall prevalence of G. duodenalis in diarrhea and nondiarrhea samples was 32.4% and 27.5%, respectively, whereas that in intensively farmed and free-range cattle was 35.0% and 19.9%, respectively. We obtained 83, 53, and 59 sequences of bg, gdh, and tpi in G. duodenalis, respectively. Moreover, assemblage A (n = 2) and assemblage E (n = 81) by bg, assemblage A (n = 1) and assemblage E (n = 52) by gdh, and assemblage A (n = 2) and assemblage E (n = 57) by tpi were identified. Multilocus genotyping yielded 29 assemblage E MLGs, which formed 10 subgroups. To the best of our knowledge, this is the first study to report cattle infected with G. duodenalis in Shanxi, China. Livestock-specific G. duodenalis assemblage E was the dominant assemblage genotype, and zoonotic sub-assemblage AI was also detected in this region.

Population Genetic Structure of the Rubber Tree Powdery Mildew Pathogen (Erysiphe quercicola) from China.

In order to manage agricultural pathogens, it is crucial to understand the population structure underlying epidemics. Rubber tree powdery mildew, caused by Erysiphe quercicola, is a serious threat to rubber plantations worldwide, especially in subtropical environments including all rubber tree-growing regions in China. However, the population structure of the pathogen is uncertain. In this study, 16 polymorphic microsatellite markers were used to genotype powdery mildew samples from the main rubber tree-growing regions including Yunnan (YN), Hainan (HN), western Guangdong (WG), and eastern Guangdong (EG). YN had higher genotypic diversity (Simpson's indices), genotypic evenness, Nei's gene diversity, allelic richness, and private allelic richness than the other regions. Cluster analysis, discriminant analysis of principal components, pairwise divergence, and shared multilocus genotype analyses all showed that YN differed significantly from the other regions. The genetic differentiation was small among the other three regions (HN, WG, and EG). Analysis of molecular variance indicated that the variability among regions accounted for 22.37% of the total variability. Genetic differentiation was significantly positively correlated (Rxy = 0.772, P = 0.001) with geographic distance. Linkage equilibrium analysis suggested possible occurrence of sexual recombination although asexual reproduction predominates in E. quercicola. The results suggested that although significant genetic differentiation of E. quercicola occurred between YN and the other regions, pathogen populations from the other three regions lacked genetic differentiation.

Clonal Outbreak of Trichophyton tonsurans Causing Tinea Capitis Among a Wrestling Team in Beijing, China.

Trichophyton tonsurans mostly causes tinea capitis and tinea corporis and often associates with outbreaks among combat sports athletes. Here, we report an outbreak of tinea capitis caused by T. tonsurans among five juvenile athletes in a wrestling team in Beijing, China. Scrapings from the lesions of the five patients were performed by direct microscopic examination and fungal culture. The fungal pathogens were all identified as T. tonsurans by morphology and sequencing of the internal transcribed spacer (ITS) regions. Multilocus genotyping analysis was performed by sequencing of 13 gene loci. The sequences of these markers were identical among the five isolates, revealing a single genotype. Antifungal susceptibilities of terbinafine, itraconazole, fluconazole, ketoconazole, and amphotericin B against T. tonsurans was determined by broth microdilution method according to the Clinical and Laboratory Standards Institute M38-A3 document and these isolates were all susceptible to the common antifungal drugs treating tinea capitis.

P309 Clonal outbreak of Trichophyton tonsurans causing tinea capitis among wrestlers in Beijing, China

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMObjectives Trichophyton tonsurans, an anthropophilic dermatophyte, mostly causes tinea capitis and tinea corporis and is often associated with outbreaks among athletes involved in combat sports, such as wrestlers and judo athletes. Here, we report an outbreak of tinea capitis caused by Trichophyton tonsurans among five juvenile athletes aged ten to fourteen in a wrestling team in Beijing, China.MethodsA total of 5 wrestlers aged 10-14 years presented with alopecia, erythema, scales, and pustule on the scalp (Fig. 1a). Scrapings from the lesions were performed by direct microscopic examination using 10% potassium hydroxide (KOH) with calcofluor white and fungal culture. The fungal pathogens were identified by morphology and sequencing of the internal transcribed spacer (ITS) regions. Multilocus genotyping analysis was performed by sequencing of the five gene loci including the ITS and non-transcribed spacer (NTS) of the ribosomal RNA (rRNA) locus, alkalineprotease-1 (ALP1), metalloprotease-5 (MEP5), carboxypeptidases Y (CarbY), which show intraspecies diversity and can be applied to epidemiological investigations and determining the route of infection transmission. Antifungal susceptibility of terbinafine (TBF), itraconazole (ITC), fluconazole (FLC), ketoconazole (KTC), and amphotericin B (AMB) against the causative fungal isolates was determined by broth microdilution method according to the Clinical and Laboratory Standards Institute (CLSI) M38-A3 document.ResultsThe five patients were diagnosed with tinea capitis because of the scalp lesions, spores, and hyphae observed by direct microscopic examination (Fig. 1b), and positive fungal culture. The causative isolates were all identified as T. tonsurans by macroscopic and microscopic characteristics (Fig. 2) and sequencing of ITS regions. The sequences of ITS, NTS, ALP1, MEP5, and CarbY were identical among the five isolates, revealing a single genotype. The abovementioned sequences have been deposited to Genbank. The minimum inhibitory concentrations (MICs) of TBF, ITC, FLC, KTC, and AMB against five T. tonsurans isolates were 0.015 μg/ml, 0.015 μg/ml, 8 μg/ml, 0.06 μg/ml, and 0.5 μg/ml, respectively.ConclusionWe report an outbreak of tinea capitis caused by T. tonsurans among wrestlers in Beijing, China. Multilocus genotyping analysis revealed that all isolates consisted of a single genotype, suggesting the outbreak may be caused by a single strain of T. tonsurans. The isolates were all susceptible to the common antifungal drugs treating tinea capitis.

Structured Populations of Critically Endangered Yellow Water Lily (Nuphar shimadai Hayata, Nymphaeaceae).

Yellow water lily (Nuphar shimadai Hayata) is a critically endangered species in Taiwan. Here, we examined genetic structures of four extant populations, WP, GPa, GPb and GPn, using 39 simple sequence repeat (SSR) markers. Positive genetic correlation was observed within 50 m, beyond which no correlation was detected due to isolation by distance according to Mantel correlogram. This suggests a significant genetic structuring of the species. Besides, multilocus genotype (MLG) analysis revealed that GPa was a panmictic population and the species’ putative center of origin. Genetic exchange was observed between GPa and GPb populations, which likely resulted from their geographic proximity. Nevertheless, there was a strong asymmetric migration detected from GPa to WP, but a recent genetic barrier prevented dispersal further northward (WP). Geneland estimated the best number of clusters as K = 2, where WP distinctly separated from the rest of the populations. In STRUCTURE output of K = 3, a third cluster was abundant only in WP. We suggest to consider GPn and WP as separate conservation units, being far from GPa. There is indeed a need to investigate these populations; as predicted, Ne = 1.6 to 3.0 is considered low and that may put the species at risk of extinction.

The Role of Genetic Factors in the Differential Invasion Success of Two Spartina Species in China.

Biological invasions have become one of the greatest threats to global biodiversity and ecosystem conservation. Most previous studies have revealed how successful invasive species adapt to new environments and climate change through phenotypic and genetic evolution. Some researchers suggested that understanding unsuccessful or less successful biological invasions might be important for understanding the relationships between invasion adaptability and climate factors. We compared the sexual reproduction ability, genetic diversity, and gene × environment interaction in two intentionally introduced alien species in China (Spartina anglica and Spartina alterniflora) based on restriction site-associated DNA (RAD) sequencing. After more than 50 years, the distribution of S. alterniflora has rapidly expanded, while S. anglica has experienced extreme dieback. A total of 212,939 single nucleotide polymorphisms (SNPs) for the two Spartina species were used for analysis. The multilocus genotype (MLG) analysis revealed that clonal reproduction was the prevalent mode of reproduction in both species, indicating that a change in the mode of reproduction was not the key factor enabling successful invasion by Spartina. All genetic diversity indicators (He, Ho, π) in S. alterniflora populations were at least two times higher than those in S. anglica populations, respectively (p < 0.001). Furthermore, the population genetic structure and stronger patterns of climate-associated loci provided support for rapid adaptive evolution in the populations of S. alterniflora in China. Altogether, our results highlight the importance of genetic diversity and local adaptation, which were driven by multiple source populations, in increasing the invasiveness of S. alterniflora.

Multilocus Genotyping of Giardia duodenalis in Alpine Musk Deer (Moschus chrysogaster) in China.

Giardia duodenalis is the underlying cause of a significant number of outbreaks of gastrointestinal illness in humans and animals worldwide. The purpose of this study was to elucidate the prevalence and genetic diversity of G. duodenalis in captive alpine musk deer (Moschus chrysogaster) in China. A total of 202 fecal samples were collected from three farms in Gansu Province, China. Identification of G. duodenalis was conducted by nested PCR targeting the genes coding for SSU rRNA, β-giardin (bg), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi). The overall prevalence of G. duodenalis in captive alpine musk deer in surveyed area was 19.3% (39/202). Two G. duodenalis genetic assemblages were identified, namely assemblage A and E. Mixed genotype infections (A+E) were found in 15.4% (6/39) of positive samples. Multilocus genotyping (MLG) analysis of G. duodenalis isolates revealed six novel assemblage A MLGs formed by two newly-described MLG-subtypes which belonged to sub-assemblage AI. To the best of our knowledge, this is the first report on MLG of G. duodenalis isolates in captive alpine musk deer in China. The presence of zoonotic assemblages and sub-assemblages of G. duodenalis in deer species suggests that these animals may potentially act as a reservoir of this protozoan for humans.

Extent of wild-to-crop interspecific introgression in grapevine (Vitis vinifera) as a consequence of resistance breeding and implications for the crop species definition.

Over the past two centuries, introgression through repeated backcrossing has introduced disease resistance from wild grape species into the domesticated lineage Vitis vinifera subsp. sativa. Introgression lines are being cultivated over increasing vineyard surface areas, as their wines now rival in quality those obtained from preexisting varieties. There is, however, a lot of debate about whether and how wine laws defining commercial product categories, which are based on the classification of V. vinifera and interspecific hybrid grapes, should be revised to accommodate novel varieties that do not fit either category. Here, we developed a method of multilocus genotype analysis using short-read resequencing to identify haplotypic blocks of wild ancestry in introgression lines and quantify the physical length of chromosome segments free-of-introgression or with monoallelic and biallelic introgression. We used this genomic data to characterize species, hybrids and introgression lines and show that newly released resistant varieties contain 76.5-94.8% of V. vinifera DNA. We found that varietal wine ratings are not always commensurate with the percentage of V. vinifera ancestry and linkage drag of wild alleles around known resistance genes persists over at least 7.1-11.5Mb, slowing down the recovery of the recurrent parental genome. This method also allowed us to identify the donor species of known resistance haplotypes, define the ancestry of wild genetic background in introgression lines with complex pedigrees, validate the ancestry of the historic varieties Concord and Norton, and unravel sample curation errors in public databases.

Multilocus genotyping analysis of 114 Giardia duodenalis isolates from different populations of domestic dogs in Japan

To determine the genotypes and zoonotic potentials of G. duodenalis isolates from different populations of domestic dogs in Japan, a total of 114 Giardia positive samples were examined using multilocus genotyping analysis at the 3 loci of glutamate dehydrogenase (gdh), beta-giardin (bg), and triose phosphate isomerase (tpi). Although the dog-adapted assemblages C and D were dominant, the zoonotic assemblage A was also demonstrated at a percentage of 23.7% (27/114). The results suggest that canine G. duodenalis isolates in Japan have the potential for zoonotic transmission

Evidence of Sexual Reproduction Events in the Dagger Nematode Xiphinema index in Grapevine Resistance Experiments Under Controlled Conditions.

The dagger nematode Xiphinema index has a major economic impact because of its transmission of Grapevine fanleaf virus to grapevines. This vector nematode, which was introduced into Western countries from the Middle East together with the domesticated grapevine, mostly reproduces by meiotic parthenogenesis, but microsatellite multilocus genotype (MLG) analysis has revealed the occurrence of rare sexual reproduction events in field conditions. In a previous 6-year study under controlled conditions, we evaluated the durability of resistance to X. index in accessions derived from a muscadine resistance source and reference accessions. In this previous study, we used an equal-proportion mixture of four lines (from Spain, Italy, Greece, and Iran) representative of X. index diversity as the inoculum, and we collected random samples in 3-, 4-, 5-, and 6-year-old vines. Here, we genotyped the individuals from these samples using the MLG technique, and we analyzed the changes in line frequency and the occurrence of sexual reproduction events between lines over time. The nematode lines differed in aggressiveness and hybrids between lines were detected at a low, but apparently increasing rate. Hybridization events were recovered in all accessions, regardless of resistance status and propagation type. Finally, our data provide the first evidence of sexual reproduction in the nematode X. index under controlled conditions.

Genetic and phenotypic effects of hybridization in independently introduced populations of the invasive maize pest &lt;em&gt;Diabrotica virgifera virgifera&lt;/em&gt; in Europe

The North American western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) was introduced into Europe several times during the end of the 20th century. Outbreaks in north-western Italy (NW Italian) and central and south-eastern Europe (CSE European) have merged in 2008 and insects interbreed since then. This study compared the genetic diversity (multi-locus genotype analyses at 13 microsatellites markers) and ten phenotypic traits among the CSE European and NW Italian populations as well as their hybrid offspring. All insects were reared under standardised laboratory conditions. Neutral genetic polymorphism appeared moderate in parental and hybrid populations, compared to North American populations. Some increase in neutral genetic variability was detected in the hybrids` expected heterozygosity and allelic richness compared to parental populations when family structures were considered. In 70% of the assessed phenotypic traits, the population type (CSE European, NW Italian, hybrids) influenced a trait, but averages in hybrids never exceeded those in their parents. Population type did not influence fecundity or adult lifespan (reflecting fitness) and not the proportion of adults flying (reflecting dispersal capabilities). There was no evidence yet that hybridization influences variability of phenotypic traits. In conclusion, there are only few indications that hybrids between the two overlapping invading European populations may in the longer term, take advantage through higher neutral genetic diversity and subsequent phenotypic adaptability.

Inferring population structure and genetic diversity of the invasive alien Nootka lupin in Iceland

Polar and subpolar regions are known for their particular vulnerability and sensitivity to the detrimental effects of non-indigenous species, which is well exemplified by the Nootka lupin (Lupinus nootkatensis) spread in Iceland. Since understanding the population and ecological genetics of invasive alien species offers hope for counteracting harmful biological invasions, the objective of the present study was to investigate interspecific variation in&nbsp;L. nootkatensis&nbsp;in Iceland in relation to a native population in Alaska. Moreover, we aimed to assess whether internal transcribed spacer 2 (ITS2) has sufficient phylogenetic applicability for a large-scale screening of the genetic diversity of a non-indigenous population of this species. This study, which is the first attempt to investigate the genetic diversity of the Nootka lupin in Iceland, included plant samples from eight locations in Iceland and one in Alaska. The analyses included genotyping by sequencing of the 417-nucleotide fragment of the 5.8S ribosomal RNA, ITS2 and part of the large subunit ribosomal RNA (GenBank MT026578-MT026580, MT077004). The main findings showed the presence of five previously unexplained single-nucleotide polymorphisms (SNPs); however, their discriminatory power for Icelandic populations was relatively low, since polymorphism information content (PIC) values ranged from 0.0182 to 0.0526, with average heterozygosity 0.0296. Concomitantly, analysis of multilocus genotypes (MLG) revealed sufficient differences in MLGs variants and their frequency to form genotypic patterns unique for Alaskan and Icelandic populations, revealing an internal genetic structure of the studied group. The proposed SNP panel needs to be supplemented with other nuclear and organellar markers.

Prevalence and multi-locus genotyping of Giardia duodenalis in rabbits from Shaanxi province in northwestern China.

Giardia duodenalis is an important parasite with veterinary and public health significance worldwide. The presence and zoonotic assemblages of G. duodenalis have previously been reported in rabbits. In this study, to understand the infection status of G. duodenalis in rabbits from Shaanxi province, a total of 537 fecal samples were collected from two breeds of rabbits in four age groups (<30 days, 31–90 days, 91–200 days and >200 days) from four geographical origins (Fengxiang, Yangling, Tongchuan, and Shanyang). The presence of G. duodenalis in these samples was assessed using molecular assays based on beta-giardin (bg). The glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) loci were then amplified in the bg-positive samples for multi-locus genotype (MLG) analysis. The total prevalence of G. duodenalis in these rabbits was 3.54% (19/537). Giardia duodenalis infection was found in both breeds of rabbits, and in all farms and age groups, but with no statistically significant differences related to these factors (p > 0.05). Two assemblages, including B and E, were identified, with the former the predominant assemblage detected in both breeds, and in all age groups and farms. Sequence analysis revealed 2 (named as rbg1-2), 1 (named as rtpi1), and 2 (named as rgdh1-2) haplotypes at the gene loci of bg, tpi, and gdh, respectively, forming a multilocus genotype (MLG) of assemblage B (rbg1, rtpi1, and rgdh1). These findings reveal the significant zoonotic potential and genetic diversity of G. duodenalis in rabbits in Shaanxi Province, PR China.

Molecular characterization of three intestinal protozoans in hospitalized children with different disease backgrounds in Zhengzhou, central China

BackgroundCryptosporidium spp. and Giardia duodenalis are major intestinal pathogens that can cause diarrheal diseases in humans, especially children. Enterocytozoon bieneusi is another parasite which can cause gastrointestinal tract disorders, with diarrhea being the main clinical symptom. However, few genetic studies of these parasites in pediatric inpatients in China have been published.MethodsTo assess the genetic characteristics and epidemiological status of these parasites, a total of 2284 fecal samples were collected from children in the pediatric departments of three hospitals in Zhengzhou, central China, and screened for these protozoans with PCR, based on the small subunit ribosomal RNA (SSU rRNA) genes of Cryptosporidium spp. and G. duodenalis and the internal transcribed spacer (ITS) of E. bieneusi.ResultsSix (0.26%), 14 (0.61%), and 27 (1.18%) of the samples were positive for Cryptosporidium spp., G. duodenalis and E. bieneusi, respectively. Of the 12 successfully sequenced G. duodenalis isolates, four were identified as assemblage A and eight as assemblage B. In subtype and multilocus genotype (MLG) analyses, C. parvum IIdA19G1 (n = 4) and two novel G. duodenalis MLGs belonging to subassemblage AII (n = 3) and BIV (n = 5) were successfully identified. The E. bieneusi isolates included genotypes D (n = 17), J (n = 2), PigEBITS7 (n = 1), BEB6 (n = 1), and CM8 (n = 1). This is the first report of C. parvum subtype IIdA19G1 in HIV-negative children and E. bieneusi genotype CM8 in humans.ConclusionsThe dominance of zoonotic C. parvum subtype IIdA19G1 indicates that this parasite is turning into zoonotic origin from human-to-human transmission. The phylogenetic analysis also revealed the zoonotic origins and anthroponotic transmission potential of G. duodenalis and E. bieneusi, suggesting more efforts must be made to minimize the threat these pathogens pose to public health.

Population Differentiation Within Anisogramma anomala in North America.

Anisogramma anomala, a biotrophic ascomycete in the order Diaporthales, causes eastern filbert blight (EFB) of hazelnuts (Corylus spp.). Until recently, little has been documented on its genetic diversity and population structure. In this study, 18 simple sequence repeat markers were used to fingerprint 182 accessions of the fungus originating from across North America. Our results, based on summary statistics of the allelic data, a discriminant analysis of principal components (DAPC) scatterplot, an unweighted pair group method with arithmetic mean (UPGMA) dendrogram, and analysis of multilocus genotypes, show that A. anomala exhibits considerable genetic diversity across multiple populations. Eleven clusters were resolved from the DAPC scatterplot, five of which were validated by statistically supported clusters in the UPGMA dendrogram. The 11 DAPC clusters were statistically significant via an analysis of molecular variance. Dendrogram topology and DAPC scatterplot groups showed some correlation with collection origin; samples collected in proximity tended to cluster together and be genetically similar. However, some locations held populations that were diverse and some populations with a high degree of similarity had disparate origins, suggesting movement by humans. Overall, the results demonstrate the presence of multiple, genetically distinct populations of A. anomala in North America and serve as a reference to assist in understanding and managing EFB.

Mating barriers between genetically divergent strains of the parasitic nematode Haemonchus contortus suggest incipient speciation

Haemonchus contortus, in common with many nematode species, has extremely high levels of genetic variation within and between field populations derived from distant geographical locations. MHco10(CAVR), MHco3(ISE) and MHco4(WRS) are genetically divergent H. contortus strains, originally derived from Australia, Kenya and South Africa, respectively, that have been maintained by numerous rounds of in vivo experimental infection of sheep. In order to explore potential pre-zygotic competition or post-zygotic incompatibility between the strains, we have investigated the ability of MHco10(CAVR) to interbreed with either MHco3(ISE) or MHco4(WRS) during dual strain co-infections. Sheep were experimentally co-infected with 4000 infective larvae (L3) per os of the MHco10(CAVR) strain and an equal number of either the MHco3(ISE) or the MHco4(WRS) strain L3. The adult worm establishement rates and the proportions of F1 progeny resulting from intra- and inter-strain mating events were determined by admixture analysis of microsatellite multi-locus genotypes. Although there was no difference in adult worm establishment rates, the proportions of F1 progeny of both the MHco10(CAVR) × MHco3(ISE) and MHco10(CAVR) × MHco4(WRS) dual strain co-infections departed from Mendelian expectations. The proportions of inter-strain hybrid F1 progeny were lower than the expected 50%, suggesting either pre-zygotic competition or post-zygotic incompatibility between the co-infecting strains. To investigate this further, both eggs and hatched L1 of broods from single adult female worms recovered from each dual co-infection were genotyped. Unhatched eggs from the broods revealed no inter-strain hybrid genotype deficit, suggesting there is no pre-zygotic competition between the strains. In contrast, there was a deficit in L1 inter-strain hybrid genotypes in the broods derived from MHco3(ISE) or MHco4(WRS) maternal parents, but not from MHco10(CAVR) maternal parents. This suggests that hybrid progeny of MHco10(CAVR) paternal parents have reduced post-zygotic development and/or viability consistent with incipient speciation of the MHco10(CAVR) strain. The presence of mating barriers between allopatric H. contortus strains has important implications for parasite ecology, including the ability of newly introduced anthelmintic-resistant parasite populations to compete and interbreed with populations already established in a region.

First report of Giardia duodenalis infection in bamboo rats

BackgroundThe zoonotic parasite, Giardia duodenalis (syns. G. lamblia and G. intestinalis), has been widely reported in humans and animals, including rodents. The bamboo rat, a rodent species belonged to the subfamily Rhizomyinae, is farmed in China because of its medicinal and edible values. However, no information of G. duodenalis infection was available in bamboo rats prior to the present study. Here, the prevalence and genetic diversity of G. duodenalis in bamboo rats from Hunan Province of China were investigated.ResultsOf 480 faecal samples collected from six farms located in four cities (Wugang, Chenzhou, Huaihua and Jishou) of Hunan Province, 52 (10.8%) were positive for G. duodenalis infection by using a nested PCR approach targeting the beta giardin (bg) gene. Significant differences (P < 0.01) in prevalence were found among different age groups and geographical localities, and among different farms in Wugang city. Sequence analysis revealed existence of the zoonotic assemblage B and genetic diversity of G. duodenalis in these animals. Multilocus genotyping analysis also indicated broad genetic diversity of assemblage B isolates in these bamboo rats.ConclusionsThis is the first report of the infection and genetic variations of G. duodenalis in bamboo rats. These findings will provide basic data for implementing effective strategies to control giardiasis in bamboo rats.

Multilocus genotype analysis outlines distinct histories for Trichinella britovi in the neighboring Mediterranean islands of Corsica and Sardinia

BackgroundThe zoonotic nematode Trichinella britovi was discovered in two neighboring Mediterranean islands of Corsica and Sardinia, almost simultaneously at the beginning of the 21st century. An epidemiological link between the two parasite populations was generally assumed. In 2015, an outbreak of trichinellosis in Nice, the South of France, was reportedly caused by the consumption of raw pork delicatessen imported from Corsica. The aims of the present study were to investigate, by multilocus genotype (MLG) analyses, the hypothesis of the common origin of the Corsican and Sardinian T. britovi foci and to trace “from fork to farm” the origin of the pork product, which caused a trichinellosis outbreak in mainland France in 2015.MethodsSixty-three T. britovi isolates were collected from animals and pork products of Sardinia and Corsica islands and from mainland of Italy, France and Spain. We analyzed genetic variability at four polymorphic microsatellite loci by two independent algorithms, the Bayesian and multivariate analyses, to evaluate the genetic relationships of 1367 single larvae.ResultsTrichinella britovi isolates of the two islands showed different genetic structures and the Bayesian analysis revealed a different membership of the two insular populations. Furthermore, two geographically separate genetic groups were identified among Corsican isolates. Lastly, the origin of the pork delicatessen marketed in Nice was linked to a breeder-butcher in Corsica.ConclusionsThe low level of genetic admixture of the insular T. britovi isolates suggests that this pathogen colonized the two islands by separate events. On the other hand in Corsica, although the isolates share the same genetic structure, geographically separate isolates showed different membership. We suggest the MLG analysis as a suitable method in supporting epidemiological investigations to trace “from fork to farm” insular populations of T. britovi.