Microsatellite Markers Research Articles

Reproductive strategies in loggerhead sea turtle Caretta caretta: polyandry and polygyny in a Southwest Atlantic rookery.

Sea turtles are highly migratory and predominantly inhabit oceanic environments, which poses significant challenges to the study of their life cycles. Research has traditionally focused on nesting females, utilizing nest counts and mark-recapture methods, while male behavior remains understudied. To address this gap, previous studies have analyzed the genotypes of females and hatchlings to indirectly infer male genotypes and evaluate the extent of multiple paternity within populations. Our research aimed to investigate the presence of multiple paternity in loggerhead turtle (Caretta caretta) nests for the first time in Brazil. We analyzed 534 hatchlings from 43 nests associated with 42 females during the 2017/18 to 2019/20 nesting seasons, using four highly polymorphic, species-specific microsatellite markers (nDNA). Parentage tests were conducted to reconstruct paternal genotypes and determine the rates of multiple paternity within clutches. Our results revealed that 72.09% of clutches were sired by multiple males, with contributions ranging from one to six males per clutch. Additionally, seven out of 88 males (7.95%) were found to have sired clutches from multiple females, with some males contributing to more than one clutch within and across breeding seasons. The breeding sex ratio (BSR) was calculated to be 2.09 males per female. While multiple paternity is a common phenomenon among sea turtles, this study is the first to document polyandry in loggerheads in Brazil and the first to provide evidence of polygyny in this species globally. This research establishes a crucial database for future studies in Brazil, with a focus on the BSR of the Southwest Atlantic subpopulation, offering essential insights for developing effective management strategies for this vulnerable population.

Microsatellite and mitochondrial DNA D-loop based analysis of genetic diversity in captive populations of Crocodylus siamensis in China

Reintroduction of threatened species is a common strategy used to mitigate population decline and reduce the risk of extinction. In order to assess the success of such reintroduction programs, it is critical to monitor the demographic and genetic features of reintroduced populations. In this work, we conducted a genetic assessment of the Critically Endangered crocodile species Crocodylus siamensis to determine the genetic diversity, utilising both microsatellite markers and mitochondrial DNA D-loop region. The genetic analysis revealed a moderate level of genetic diversity within the sampled populations, as indicated by the microsatellite markers (Ho = 0.505 ± 0.080; He = 0.297 ± 0.091) and mtDNA D-loop (hd = 0.622 ± 0.061; Pi = 0.01294 ± 0.00041). Notably, 22 distinct haplotypes were observed. Additionally, we identified evidence of hybridisation that might have occurred between C. siamensis and Crocodylus niloticus raising concerns about the potential compromise of the species' genetic integrity. This study represents the first genetic assessment of captive bred C. siamensis populations in China, encompassing both microsatellite and mtDNA D-loop analyses. To ensure preservation of C. siamensis, we recommend identifying and excluding hybrid individuals from conservation programmes. Our findings support the feasibility of implementing breeding and captive management plans for C. siamensis, providing a solid theoretical basis for their reintroduction and captive breeding to safeguard and augment the population of this Critically Endangered freshwater crocodile. This research marks the initial step towards establishing a scientifically sound source population for the conservation of C. siamensis in China. Keywords: protection, reintroduction genotype, haplotypic diversity, nucleotide diversity

Landscape and conservation genetics of western black crested gibbons (Nomascus concolor) in China.

Despite decades of field study, very little is known about the molecular ecology of gibbons, particularly as it relates to their ability to disperse across degraded and fragmentary landscapes. The critically endangered western black crested gibbon (Nomascus concolor) has been reduced to a small, fragmented population with about 1300 individuals. In the largest population genetic study of free-ranging gibbons to date, we sampled 47 of these gibbons from 13 sites in China and generated 15 polymorphic autosomal microsatellite markers. We identify three population clusters of N. concolor in Yunnan centered in 1) the Wuliang and Ailao Mountains, 2) the Yongde Daxueshan Mountains, and 3) an isolated remnant near the border with Vietnam. Within the Wuliang Mountains, we identified four subclusters, three of which are bounded by high-altitude rhododendron forest, and one that is isolated from the main population by ~2 km of degraded forest and pasture. Least-cost path analysis and isolation by resistance modeling demonstrates that the population genetic distances among gibbons in Wuliangshan National Nature Reserve are significantly correlated with geographic paths that avoid use of high-altitude rhododendron forest in favor of evergreen broadleaf forest. Although these gibbons have likely undergone reductions in heterozygosity from recent consanguineous mating, we suggest that their active avoidance of inbreeding on the population level maintains higher than expected levels of genetic diversity. This research provides new insights into how gibbons interact with heterogeneous environments and expands our understanding of their molecular ecology and conservation genetics.

Genetic structure and demographic analysis of a true single-population species, Camellia azalea

Single-population species (SPS) consist of only one natural population and often are at high risk of extinction. Although almost all species must go through this special stage in their evolutionary process, there is little understanding of how SPS survives. Camellia azalea C. F. Wei is a typical SPS, and has precious breeding values for its special flowering period. This study surveyed the age structure and spatial distribution of C. azalea, analyzed its genetic diversity and fine-scale spatial genetic structure (SGS) using microsatellite markers for 629 individuals, and estimated the effect of human disturbances on its population dynamics. Results showed that this species had a relatively moderate genetic diversity (I = 0.989, He = 0.509, and Ho = 0.497), high rate of sapling (~ 35%), and a narrow habitat (~ 6 km long, ~ 10 m wide). Although the construction of dams and roads did not lead to a significant loss of genetic diversity and genetic differentiation (FST = 0.0096 ~ 0.0128, Nm = 19 ~ 26), it limited C. azalea’s seed flow (adults, 95 m; juveniles, 60 m), which was a reason for juveniles having a stronger SGS than adults. These results indicate that as an SPS, C. azalea still possesses the potential capacity for self-evolution and regeneration, however, it is at risk of extinction due to its small range, narrow habitat, and human distances. Furthermore, the results are also of enlightening significance to the conservation of other SPS, especially those distributed along the riparian zone.

Impacts of habitat fragmentation on the genetic diversity of the endangered Guatemalan fir (Abies guatemalensis Rehder).

Abies guatemalensis Rehder, an endangered conifer endemic to Central American highlands, is ecologically vital in upper montane forests. It faces threats from habitat fragmentation, unsustainable logging, and illegal Christmas tree harvesting. While previous genetic studies on mature trees from eighteen populations showed high within-population diversity and limited among-population differentiation, the genetic impact of recent anthropogenic pressures on younger generations has yet to be discovered. Understanding these effects is crucial for developing effective conservation strategies for this vulnerable species. We sampled 170 young trees (< 15 years old) from seven populations across Guatemala. Seven microsatellite markers were used to analyse genetic diversity, population structure, and recent demographic history. Moderate levels of genetic diversity were observed within populations (mean Shannon diversity index = 4.97, mean Simpson's index = 0.51, mean allelic richness = 11.59, mean observed heterozygosity = 0.59). Although genetic structure broadly aligned with mountain corridors, substantial admixture patterns suggest historical connectivity across all populations. Most populations showed evidence of recent bottlenecks (p < 0.05) and inbreeding. The results suggest a potential decline in genetic diversity and increased population structuring (ΦST = 0.274, p < 0.01) over the past decades compared to the previous study on old trees. The observed genetic patterns indicate ongoing impacts of habitat fragmentation and anthropogenic pressures on A. guatemalensis. Conservation efforts should prioritise expanding effective population sizes and facilitating gene flow, particularly for isolated populations. While restoration efforts may be logistically easier within mountain ranges, genetic evidence suggests that increasing overall population connectivity could benefit this species. Management strategies should implement systematic seed collection protocols to maintain genetic diversity in future populations. These findings highlight the urgent need for conservation measures to preserve remaining genetic diversity and promote connectivity among A. guatemalensis populations.

Genetic Diversity and Structure of Higher-Resin Trees of Pinus oocarpa Schiede in Mexico: Implications for Genetic Improvement

Pinus oocarpa Schiede is the most widely distributed conifer in the Americas. In Mexico, it inhabits diverse environments and is the primary pine species utilized for resin production, prompting the establishment of a genetic improvement program (GIP). Preserving a broad genetic diversity is fundamental to the success of the GIP. This study aimed to assess the genetic diversity and structure of trees selected for their high resin yield. A total of 146 trees from 15 provenances within three populations (MX-MIC, MX-MEX, and MX-OAX) constituting the selection population of the GIP were evaluated. Five SSR microsatellite markers (PtTX3013, NZPR1078, PtTX2146, PtTX3107, and PtTX3034) were used to determine key indicators of genetic diversity and structure. All three populations exhibited high genetic diversity; however, the heterozygosity observed was lower than the expected heterozygosity. Genetic structure analysis revealed the presence of two distinct genetic groups: the Transverse Volcanic Axis (MX-MIC and MX-MEX) and the Sierra Madre del Sur (MX-OAX). Most of the genetic diversity (87.42%) was found within provenances. Gene flow was high among provenances within the same genetic group but limited between provenances from different groups. The findings suggest that GIPs should be tailored to each genetic region, with a focus on within-provenance selection to maintain genetic diversity.

The influence of post-glacial migration and hybridization on the gene pool of marginal Quercus pubescens populations in Central Europe.

In Central Europe, the drought-tolerant downy oak (Quercus pubescens) is at the northern edge of its natural distribution range, often growing in small and spatially isolated populations. Here, we elucidate how the population genetic structure of Central European Q. pubescens was shaped by geographic barriers, genetic drift and introgression with the closely related sessile oak (Q. petraea). 27 Q. pubescens populations from the northern margin of Q. pubescens' natural distribution range were sampled. Based on 16 nuclear microsatellite markers (nSSRs), Bayesian clustering and distance-based analyses were performed to determine the intraspecific genetic structure and to identify genetic barriers. To identify drivers of introgression with Q. petraea, generalised linear models were applied to link levels of introgression with environmental conditions. To track post-glacial migration routes, the spatial distribution of haplotypes based on 8 chloroplast microsatellite markers (cpSSRs) was investigated. Based on nSSRs, the study populations of Q. pubescens were divided into a western and an eastern genetic cluster. Within these clusters, more pronounced genetic substructure was observed in the west, probably due to a rugged topography and limited gene flow. Introgression from Q. petraea was more prevalent at wetter and north-exposed sites and in the west. The identified cpSSR haplotypes followed known migration pathways. Our results suggest two late-glacial refugia in or near the southwestern Alps and the southeastern Alps as potential sources for post-glacial migration. Although some genetic exchange is evident in Northern Italy, south of the Alps, the two clusters remain distinct at a large scale. Landscape features and introgression with Q. petraea shaped the genetic substructure at a smaller scale. Our study provides a comprehensive overview of the genetic structure of Q. pubescens in Central Europe, relevant for conservation.

Genome-Wide In Silico Analysis of Microsatellite Loci in Rabbits.

This study aimed to characterize microsatellites in the rabbit genome using an in silico approach and to develop and validate microsatellite markers. Blood samples were collected from 15 Baladi rabbits and 18 New Zealand White (NZW rabbits). The GMATA software was used to define SSRs in the extracted sequences. Twelve primer pairs were used to validate the loci identified and the primers developed. The total number of the detected microsatellite loci overall chromosomes was 1,136,253. The di-nucleotide microsatellite repeats dominated and exceeded 88% of the detected microsatellites in all chromosomes. There were no microsatellites detected in mitochondrial DNA. The highest relative microsatellite abundance was obtained for chromosome 19, followed by 13 and 6. The highest estimated SSR density was obtained for chromosome 14, and the lowest was for mitochondrial DNA, followed by chromosome 13. The polymorphism was 81.63% and 75.51% for Baladi and NZW rabbits, respectively. The number of detected alleles ranged between two and seven alleles/loci, and polymorphic information content was from 35% to 71%. The AMOVA analysis showed that the total variance of all levels of population structure was 15.734. The results definitely confirmed higher genetic diversity in Baladi compared with NZW rabbits.

Unearthing Genetic Treasures: Exploring Lost Autochthonous Vitis vinifera Varieties in Lebanon.

Lebanon, one of the oldest centers of grapevine (Vitis vinifera L.) cultivation, is home to a rich diversity of local grape varieties. This biodiversity is linked to the country's unique topography and millennia of cultural history. However, the wine industry primarily utilizes international varieties, putting many local varieties at risk of extinction. In this study, we analyzed 202 samples from old vineyards, home gardens, and private collections using 21 microsatellite markers to assess their identity and genetic diversity. A total of 67 different genotypes were identified, with 34 not matching any existing profiles in the consulted databases, based on comparisons with the European Vitis Database, the Vitis International Variety Catalogue (VIVC), and the databases established in two previous studies conducted in Armenia and Lebanon. Cluster analyses revealed Lebanon's rich diversity of local grape varieties, highlighting cases of synonymy, homonymy, and misnaming. All loci were polymorphic, with 228 alleles and an average of 11.4 alleles being detected. The highest number of alleles was observed at the VVIV67 locus (19 alleles), while the lowest was found at the VVIQ52 and VVIN73 loci (5 alleles). The observed heterozygosity was 0.732, slightly below the expected value of 0.757, with gene diversity varying among the markers. Of the 67 genetic profiles identified, 34 are absent from national and international databases, underscoring Lebanon as a hotspot for grapevine genetic diversity. This unique genetic variation, which includes several synonyms due to geographic isolation, could provide valuable opportunities for producing distinctive wines and emphasizes the need for further research and documentation.

Increasing Beef Production in the Northern Region of the Republic of Kazakhstan Using the Genetic Resources of Aberdeen Angus Cattle of Different Genotypes.

This article presents the findings of a scientific study investigating the efficacy of various assessment techniques used to evaluate the adaptability and productive qualities of Aberdeen Angus cattle on three prominent farms in the northern region of the Republic of Kazakhstan. A comprehensive analysis of the haematological and biochemical parameters of experimental groups of cattle with different genotypes (American, Canadian, and Estonian selection) was conducted. The studies revealed notable variability in haematological and biochemical indicators, contingent on the origin. Concurrently, the dynamics of the aforementioned indicators did not exceed the physiological norms. The modern allelofund was characterised with the help of microsatellite markers, and the level of genetic diversity of Aberdeen Angus cattle of different genotypes was estimated. The research uncovered the genealogical structure of the populations, the purity of the populations, the provenance, the polymorphism level, the heterozygosity indices, and the Wright fixation index (Fis). The genotyping of cattle of the Aberdeen Angus breed on 15 microsatellite markers yielded the establishment of 80 alleles in the Kolos-firm LLP, 77 alleles in the Vishnevskoe LLP, and 92 alleles in the Sever-Agro N LLP. The total expected heterozygosity was He = 0.673, while the observed heterozygosity was Ho = 0.710.

Microsatellite markers development and molecular fingerprinting of cashew cultivars.

Cashew (Anacardium occidentale L.) is a widely cultivated tree with great economic significance. In cashew, several elite cultivars have been developed for commercial cultivation, which form the underpinning for the cashew-based industries and the several billion-dollar world trade. However, frequently the genetic purity of the planting material is not maintained, resulting in great economic losses. Therefore, there is a need to develop a reliable method for the identification of cultivars to avoid economic losses to the cultivators and the protection of cultivars by the breeders. In this study, 35 new microsatellite/simple sequence repeat (SSR) markers were developed, and a set of 20 highly polymorphic and reproducible markers were used for DNA fingerprinting and genetic diversity analysis in 32 cashew cultivars. The polymorphic information content (PIC) of newly developed markers varied from 0.19 to 0.67, with an average of 0.44. The probability of identifying any two genotypes with identical fingerprints using the 20 SSR markers used for fingerprinting here in cashew cultivars was less than 2.8 × 10-11. Of the set of 20 markers, eight were sufficient for uniquely fingerprinting all the cultivars. Genetic diversity analysis by the neighbor-joining (NJ) method grouped 32 cultivars into three main clusters, and the grouping had no relationship to the geographic regions or the pedigree. The findings of this study are useful for the conservation and protection of cultivars under the PVP Act for ensuring the trading of quality planting material and are also useful for cashew breeding programs.

Novel Efficient Genome-Wide STR Panels for the Conservation of the Endemic Catfishes of Western Ghats: Critically Endangered Hemibagrus punctatus Jerdon (1849) and Vulnerable Horabagrus brachysoma (Günther, 1864).

The IUCN classifies two-endemic fishes of the Western Ghats in Peninsular India: Nilgiri mystus Hemibagrus punctatus is critically endangered, and Yellow catfish Horabagrus brachysoma is vulnerable. These are threatened by habitat degradation, excess fishing, and dam construction. ICAR-NBFGR, the premier research organisation in India for fish conservation, aims to conserve these two species through captive propagation, stock identification, and stock-specific ranching programs. The information on this species' genetic diversity and population structure is crucial for the successful rehabilitation of the species. Microsatellite markers are considered suitable markers for genetic stock structure analyses. An attempt was made to identify microsatellite markers in H. brachysoma (n = 27) and H. punctatus (n = 31) using Illumina Sequencing technology. The preliminary studies show that markers are efficient enough to differentiate the genetic stocks of these conservation-important species across their range of natural distribution. Three genetic stocks are identified in H. brachysoma and two in H.punctatus using the developed markers.

Population Structure, Morphology, and Health Assessment of Philippine Swamp Buffalo (Bubalus kerabau, Fitzinger, 1860) in Calayan Island, Cagayan

Understanding the state of the swamp buffalo population in Calayan Island is important to strengthen the conservation and management program in the country. This study aimed to provide insights into the morphology, population structure, and health profile of the swamp buffaloes on the island. In total, 35 fresh blood samples were analyzed using 27 polymorphic microsatellite markers to determine the population structure analysis. Data were gathered for the morphological features of Calayan swamp buffaloes and served as the baseline information for the descriptive traits. Furthermore, samples were tested for surra and brucellosis using PCR and serological tests, respectively. The results showed that Calayan swamp buffaloes were morphologically bigger, except for body length, compared with other populations. A genotype analysis using microsatellite markers showed remarkable discriminatory power to distinguish distinct populations within the tested population, and could discriminate subspecies of swamp and river types plus crossbreds. The study also reports the first incidence of surra and brucellosis on the island. Overall, the new insights provided on the newly detected Philippine carabao lineage in Calayan Island would be highly recommended for the ex situ conservation and animal health control strategy. The conservation strategy would encompass collecting, cryopreserving, and storing viable germplasms from local swamp buffalo on Calayan Island.

Preserving tororaro’s legacy: conservation genetics of in-situ and ex-situ populations of Muehlenbeckia astonii (Polygonaceae)

ABSTRACT Tororaro, a Threatened Nationally Endangered species in eastern South Island and southern North Island, New Zealand, is primarily found in coastal or lowland droughty habitats with small, isolated populations, notably the largest on Kaitōrete Spit, Canterbury. Due to recent issues with the conservation management of the Kaitōrete Spit population, the importance of protecting this species has been highlighted. While widely cultivated in gardens for its horticultural appeal and resilience, recruitment failure in wild populations and the genetic structure and variation within wild and cultivated populations is unknown, which raises questions about in-situ and ex-situ conservation strategies. Microsatellite markers were developed to assess the level of natural variation present in wild populations and that of cultivated material present throughout New Zealand. The results reveal natural populations can be grouped into three clusters with a low to medium level of diversity for each cluster (Na = 1.647–3.647, Ho = 0.194–0.457, He = 0.181–0.478), and the diversity of each group is captured in cultivation (Na = 2.471–3.588, Ho = 0.207–0.511, He = 0.173–0.519). Cultivated material largely follows ecosourcing guidelines whereby cultivated plants in Wellington, Marlborough and Canterbury are often sourced from the regional meta-population, however, there are some exceptions that have implications for management of this species. In Otago, where M. astonii does not naturally occur, cultivated populations comprise representatives of all regional populations. An interesting outcome is that we recovered from among the cultivated plants several individuals that are likely to be putative intraspecific hybrids between plants from Wellington populations and those from either Marlborough or Canterbury; these hybrid genotypes are unknown in wild populations. Clonal plants were also represented among the wild and cultivated populations by individual plants having identical genotypes. We also demonstrate the utility of these markers across other Muehlenbeckia species.

Association of Microsatellite Markers on Chromosome Seven with Milk Production and Fertility Traits in Egyptian Buffalo

Association of Microsatellite Markers on Chromosome Seven with Milk Production and Fertility Traits in Egyptian Buffalo

Evidence of sterility of the male sporophytes of the brownalga Saccharina japonica (Phaeophyceae) in cultureirrespective of their ploidy levels.

Monoclonal female gametophytes of Saccharina japonica, when cultured independently, can develop into female sporophytes. Previous research has shown that the chromosomes in female sporophytes of S. japonica may naturally duplicate, forming diploids, and these diploid female sporophytes are capable of forming sori and releasing zoospores. In contrast, male sporophytes derived from culturing monoclonal male gametophytes failed to form sori and produce meiospores; thus, it was hypothesized that these male sporophytes are haploid and unable to perform meiotic division. In order to explore whether the chromosomes in male sporophytes can naturally duplicate and whether such diploid male sporophytes can effectively form sori and produce meiospores, we obtained 62 male sporophytes and cultivated them into adults. Only male-specific DNA markers were detected in all of these male sporophytes, indicating their male nature. Ten microsatellite markers were used to estimate the relatedness of the parental gametophytes and the corresponding sporophytes. Results revealed that the genotypes of the male sporophytes matched exactly with the corresponding male gametophytes. Both diploid and haploid sporophytes were detected in these 62 male sporophytes when analyzed by flow cytometry. After 16 months of cultivation, none of the male sporophytes formed sori regardless of their ploidy. As controls, both female and hybrid sporophytes developed sori and released viable zoospores. These findings suggest that the sterility of male sporophytes in S. japonica is not related to their ploidy; rather, it is attributed to the absence of essential elements for sori formation that may be present only in the female.

The spread of molecular markers of artemisinin partial resistance and diagnostic evasion in Eritrea: a retrospective molecular epidemiology study

Eritrea was the first African country to discontinue the use of histidine rich protein 2 (HRP2)-detecting rapid diagnostic tests (RDTs) for malaria diagnosis following reports of a high prevalence of pfhrp2/3-deleted Plasmodium falciparum parasites causing false-negative results in the country. Eritrea was also the first African country to report partial artemisinin resistance due to the Pfalciparum kelch13 (pfk13) Arg622Ile mutation. We aimed tocharacterise the spatial distribution of pfk13 mutants and their interactions with pfhrp2/3 deletions in Eritrea andto assess the role of the use of HRP2-detecting RDTs and antimalarial (artesunate-amodiaquine) therapy in the spread of the two variants. We conducted a retrospective molecular epidemiological analysis of pfk13 mutations and pfhrp2/3 deletions in existing Pfalciparum-infected blood samples collected as part of previous pfhrp2/3 deletion and severe malaria studies. Samples were collected in March, 2016and between September, 2018, and January, 2020, from symptomatic patients seeking care at 15health centres in four administration zones (Semenawi Keyih Bahri, Gash Barka, Anseba, and Debub) in Eritrea. Afragment spanning the propeller region of pfk13 was amplified from samples and sequenced using Sanger sequencing or targeted amplicon sequencing to identify genetic mutations. Deletions of pfhrp2/3 genes in samples were determined using multiplex quantitative PCR. Parasite haplotypes and genetic relatedness of parasite haplotypes were determined previously using microsatellite marker typing. The primary objective was to determine the prevalence of pfk13 mutations at health centres and administrative zones. The secondary objective was to investigate whether pfk13 mutants and pfhrp2/3 deleted parasites converge. We sequenced 50samples collected in March, 2016from the Semenawi Keyih Bahri zone and identified no pfk13 mutations. By contrast, in 587samples included in this study that were collected from health centres in GashBarka, Anseba, and Debub in 2018-20, we detected five different single non-synonymous mutations: Glu605Lys, Arg622Ile, Asn657Lys, Lys658Glu, and Ser679Leu. The most prevalent mutation was pfk13 Arg622Ile, which was detected in samples collected from all nine health centres where more than five samples were available across all three administration zones, with an overall prevalence of 11·9% (70 of 587samples; range 5·9-28·0%). Weidentified 22unique pfk13 Arg622Ile mutant haplotypes among 26samples tested, of which 13 (59·1%) were genetically related, whereas the remaining nine (40·9%) were not. The prevalence of pfk13 Arg622Ile was significantly higher in parasites with a single pfhrp3 deletion (46 [18·0%] of 255samples) than in parasites withoutpfhrp2/3 deletions (ten [6·2%] of 161samples; odds ratio 3·89 [95% CI 1·59-7·61]; p=0·0006) and with dual pfhrp2/3-deleted parasites (13 [9·0%] of 145; 2·23 [1·13-4·68]; p=0·018). The geographical spread of the pfk13 Arg622Ile mutation might have initially resulted from the clonal expansion and spread of pfhrp2/3 deletions under the test-and-treat policy using HRP2-detecting RDTs. Subsequently, selective pressure from artemisinin combination therapy could have further facilitated the spread of both pfk13 Arg622Ile and pfhrp2/3 deletions. Continuous monitoring of trends in pfk13 and pfhrp2/3 variants is needed to inform effective malaria control and elimination strategies in Eritrea and other African countries. US Department of Defense Armed Forces Health Surveillance Division, Global Emerging Infections Surveillance Branch (AFHSD/GEIS), and Wellcome Trust.

ECHINOCOCCUS MULTILOCULARIS IN NEW YORK WILDLIFE: DISTRIBUTION AND GENETIC DIVERSITY OF AN EMERGING PATHOGEN.

Echinococcus is a genus of cestode parasites of paramount veterinary and medical importance globally. Two species, Echinococcus granulosus sensu lato and Echinococcus multilocularis, are endemic to North America and are the etiologic agents of cystic echinococcosis and alveolar echinococcosis, respectively. North America is currently experiencing an epidemiological shift in the state of transmission, distribution, and prevalence of E. multilocularis, following the introduction of the European variant of the parasite. The rapid expansion of the parasite into the northeastern United States has resulted in novel detections of spillover to humans, indicating the need for active surveillance to direct public health education and disease control measures. Our surveillance of the parasite in definitive hosts in New York State from 2019 to 2023 yielded 8 detections of the parasite, for a sample prevalence of 8.3% (8/96). Phylogenetic analysis of mitochondrial markers confirmed that all New York isolates belong to the European lineage and group in a well-supported clade with E. multilocularis isolates described from Slovakia and Poland. Investigation of the highly polymorphic EmsB microsatellite marker revealed several E. multilocularis genotypes circulating in New York, including 1 that is distinct from European isolates detected in western North America. These data are consistent with 1 or more unique introductions from Europe rather than a spread from western regions of Canada and the United States, where other European variants are found. Echinococcus multilocularis appears to have a complex epidemiology in North America that warrants further detailed surveillance to evaluate the risks for zoonotic transmission.

The use of microsatellite markers for assessing genetic identity of fruit tree crop plant cultivars

The use of microsatellite markers for assessing genetic identity of fruit tree crop plant cultivars

Systemic inflammatory response in colorectal cancer is associated with tumour mismatch repair and impaired survival

The systemic inflammatory response (SIR), defined as elevated levels of circulating C-reactive protein (CRP), is an important predictor of impaired survival in colorectal cancer. The aim of this study was to explore the prognostic role of SIR and its association with tumour mismatch repair status and the immune response. Immune activity profiles of mononuclear cells isolated from CRC tissues and blood in the U-CAN exploration cohort (n = 69), were analysed by flow cytometry. In the U-CAN validation cohort (n = 257), T-helper cells (T-bet+), cytotoxic T cells (CD8+), regulatory T cells (FoxP3+), B cells (CD20+), and macrophages (CD68+) were analysed by multispectral imaging. Microsatellite instability was determined using five mononucleotide-repeat microsatellite markers. Patients with high CRP levels (> 10 mg/l) were significantly more often diagnosed with high-grade tumours and tumours exhibiting microsatellite instability. However, some patients with high CRP levels were found to have microsatellite-stable tumours. Furthermore, high CRP levels were associated with specific tumour immune traits including an augmented macrophage response and were significantly linked to poorer cancer-specific survival, particularly in patients with microsatellite-stable tumours. In conclusion, our findings suggest an interplay between SIR and mismatch repair status in CRC prognosis which needs to be further explored.