Recombinase-aid amplification combined with lateral flow detection assay for sex identification of the great white pelican (Pelecanus onocrotalus).

Sex identification in avian species is essential for biodiversity conservation and ecological studies. However, the sex of nearly half of the birds could not be identified based on their external appearance. It is difficult to visually identify sex to monitor the ecology and conservation of wild populations. In this study, we designed primer pairs for large white pelican using recombinase-based isothermal amplification combined with a lateral flow dipstick (RAA-LFD) assay for chromo-helicase-DNA binding protein (CHD) genes mapped to W chromosomes and an ultra-conserved element (UCE) located on chromosome 6, respectively. Our result showed that the raaW4-RAA-LFD can detect up to 0.1 ng of genomic DNA (gDNA) templates of female pelicans in 30 min at 39 ℃ and accurately distinguish female from male without any cross reactivity. RaaUCE2-RAA-LFD can amplify both male and female pelicans with a detection limit of 25 pg. To further evaluate the assay, 15 white pelicans of unknown sex were tested using the RAA-LFD assay and conventional polymerase chain reaction (PCR). The results of the raaW4-RAA-LFD assay were consistent with those of the conventional PCR. The developed RAA-LFD assay is equipped with field-deployable instruments and offers a field platform for rapid and reliable sex identification in pelicans.

Development of DNA Insertion-specific Markers Based on the Intron Region of Oplegnathus punctatus itih4b for Genetic Sex Identification.

Spotted knifejaw (Oplegnathus punctatus) is a significant marine fish species that exhibits pronounced sexual dimorphism, with males generally exhibiting greater weight and growth rates than females. Therefore, the farming of O. punctatus with a high proportion of males is beneficial for improving the quality and efficiency of the O. punctatus aquaculture industry. Furthermore, the development of a rapid technique in sexing O. punctatus fry will facilitate the selection and breeding of superior male varieties of O. punctatus. In this study, genome-wide scanning, comparative genomics, and structural variation analysis methods were employed to identify and extract the homologous region of the inter-alpha-trypsin inhibitor heavy chain 4 (itih4b) gene on the X and Y chromosomes from the complete genome sequence of O. punctatus. This analysis revealed the presence of a large segment of DNA insertion markers on the Y chromosome in the region. Itih4b plays an important role in the mechanisms that regulate inflammatory and immune responses in multicellular organisms. The method described here involved the design of a pair of primers to amplify two bands of 532 bp and 333 bp in males (individuals with DNA insertion variants in the intron of the itih4b gene). In females (individuals without DNA insertion), only one band of 333 bp could be distinguished by agarose gel electrophoresis. This method shortened the time required to accurately characterize intronic DNA insertion variants and genetic sexes in O. punctatus, thereby improving detection efficiency. This study has significant value for the large-scale breeding of O. punctatus all-male seedlings and provides a reference point for the study of intron variation regulation and RNA shearing in the itih4b gene.

Development of Reliable Male-Specific Molecular Markers for Genetic Sex Identification in Sea Cucumber Apostichopus japonicus.

Apostichopus japonicus is an important marine aquaculture species in China, with high nutritional and economic value. In A. japonicus, there is no obvious sexual dimorphism in external appearance, and sex differentiation primarily relies on the observation of mature gonads after dissection, which leads to difficulties in sex identification. The confusion in sex identification greatly reduces breeding efficiency in the sea cucumber industry. Therefore, developing a reliable sex-specific marker is crucial. In this study, we identified 586 male-specific sequences through whole-genome assembly and sequence alignment, but did not identify any female-specific sequences, inferring an XY-type sex determination system in sea cucumbers. We developed a set of male-specific molecular markers to establish an accurate, stable, and widely adaptable genetic sex identification technique for A. japonicus. The male-specific molecular markers were validated with 100% accuracy in sea cucumber populations from six different geographical regions in China. In conclusion, this study provides further evidence for the XY-type sex determination system in A. japonicus and establishes an effective genetic sex identification method for multi-geographic populations, which benefits future study on reproductive biology and has significant implications in sea cucumber aquaculture industry.

Sex-specific diet differences in harbor seals (Phoca vitulina) via spatial assortment.

The lack of recovery of Chinook salmon (Oncorhynchus tshawytscha) in the Pacific Northwest has been blamed in part on predation by pinnipeds, particularly the harbor seal (Phoca vitulina). Previous work at a limited number of locations has shown that male seal diet contains more salmon than that of female seals and that sex ratios at haul-out sites differ spatiotemporally. This intrapopulation variation in predation may result in greater effects on salmon than suggested by models assuming equal spatial distribution and diet proportion. To address the generality of these patterns, we examined the sex ratios and diet of male and female harbor seals from 13 haul-out sites in the inland waters of Washington State and the province of British Columbia during 2012-2018. DNA metabarcoding was conducted to determine prey species proportions of individual scat samples. The sex of harbor seals was then determined from each scat matrix sample with the use of quantitative polymerase chain reaction (qPCR). We analyzed 2405 harbor seal scat samples using generalized linear mixed models (GLMMs) to examine the factors influencing harbor seal sex ratio at haul-out sites and permutational multivariate analysis of variance (PERMANOVA) to examine the influence of sex and haul-out site on harbor seal diet composition. We found that the overall sex ratio was 1:1.02 (female:male) with notable spatiotemporal variation. Salmoniformes were about 2.6 times more abundant in the diet of males than in the diet of females, and Chinook salmon comprised ca. three times more of the average male harbor seal's diet than the average female's diet. Based on site-specific sex ratios and diet data, we identified three haul-out sites where Chinook salmon appear to be under high predation pressure by male harbor seals: Cowichan Bay, Cutts Area, and Fraser River. Our study indicates that combining sex-specific pinniped diet data with the sex ratio of haul-out sites can help identify priority sites of conservation concern.

Development of DNA Insertion-specific Markers Based on the Intergenic Region of Oplegnathus punctatus Cdkn1/srsf3 for Sex Identification.

Spotted knifejaw (Oplegnathus punctatus) is a marine economic fish with high food and ecological value, and its growth process has obvious male and female sexual dimorphism, with males growing significantly faster than females. However, the current sex identification technology is not yet mature, which will limit the growth rate of O. punctatus aquaculture and the efficiency of separate sex breeding, so the development of efficient sex molecular markers is imperative. This study identified a 926 bp DNA insertion fragment in the cdkn1/srsf3 intergenic region of O. punctatus males through whole-genome scanning, comparative genomics, and structural variant analysis. A pair of primers was designed based on the insertion information of the Y chromosome intergenic region in male individuals. Agarose gel electrophoresis revealed the amplification of two DNA fragments, 1118 bp and 192 bp, in male O. punctatus individuals. The 926 bp fragment was identified as the insertion in the intergenic region of cdkn1/srsf3 in males, while only a single 192 bp DNA fragment was amplified in females. The biological sex of the individuals identified in this manner was consistent with their known phenotypic sex. In this study, we developed a method to detect DNA insertion variants in the intergenic region of O. punctatus. Additionally, we introduced a new DNA marker for the rapid identification of the sex of O. punctatus, which enhances detection efficiency. The text has important reference significance and application value in sex identification, all-male breeding, and lineage selection. It provides new insights into the regulation of variation in the intergenic region of cdkn1/srsf3 genes and the study of RNA shearing.

Ginkgo biloba Sex Identification Methods Using Hyperspectral Imaging and Machine Learning.

Ginkgo biloba L. is a rare dioecious species that is valued for its diverse applications and is cultivated globally. This study aimed to develop a rapid and effective method for determining the sex of a Ginkgo biloba. Green and yellow leaves representing annual growth stages were scanned with a hyperspectral imager, and classification models for RGB images, spectral features, and a fusion of spectral and image features were established. Initially, a ResNet101 model classified the RGB dataset using the proportional scaling-background expansion preprocessing method, achieving an accuracy of 90.27%. Further, machine learning algorithms like support vector machine (SVM), linear discriminant analysis (LDA), and subspace discriminant analysis (SDA) were applied. Optimal results were achieved with SVM and SDA in the green leaf stage and LDA in the yellow leaf stage, with prediction accuracies of 87.35% and 98.85%, respectively. To fully utilize the optimal model, a two-stage Period-Predetermined (PP) method was proposed, and a fusion dataset was built using the spectral and image features. The overall accuracy for the prediction set was as high as 96.30%. This is the first study to establish a standard technique framework for Ginkgo sex classification using hyperspectral imaging, offering an efficient tool for industrial and ecological applications and the potential for classifying other dioecious plants.

Identification of Sex-Associated Genetic Markers in Pistacia lentiscus var. chia for Early Male Detection.

Pistacia lentiscus var. chia is a valuable crop for its high-added-value mastic, a resin with proven pharmaceutical and cosmeceutical properties harvested from the male tree trunk. To achieve the maximum economic benefits from the cultivation of male mastic trees, it is important to develop early sex diagnosis molecular tools for distinguishing the sex type. Thus far, the work on sex identification has focused on Pistacia vera with promising results; however, the low transferability rates of these markers in P. lentiscus necessitates the development of species-specific sex-linked markers for P. lentiscus var. chia. To our knowledge, this is the first report regarding: (i) the development of species-specific novel transcriptome-based markers for P. lentiscus var. chia and their assessment on male, female and monoecious individuals using PCR-HRM analysis, thus, introducing a cost-effective method for sex identification with high accuracy that can be applied with minimum infrastructure, (ii) the effective sex identification in mastic tree using a combination of different sex-linked ISSR and SCAR markers with 100% accuracy, and (iii) the impact evaluation of sex type on the genetic diversity of different P. lentiscus var. chia cultivars. The results of this study are expected to provide species-specific markers for accurate sex identification that could contribute to the selection process of male mastic trees at an early stage for mass propagation systems and to facilitate future breeding efforts related to sex-linked productivity and quality of mastic resin.

New primers for sex identification in the Chukar partridges (Alectoris chukar).

1. Male and female Chukar partridges are difficult to differentiate based on their morphology or by the Chromobox-Helicase-DNA binding (CHD) during early growth.2. The current study developed a novel, simple, low-cost and rapid sexing protocol for Chukar partridges based on the newly defined sexing gene ubiquitin-associated protein 2 (UBAP2).3. The length of polymorphism between UBAP2-W and UBAP2-Z homologous genes allows for easy sex discrimination in this species. Molecular sexing analysis was based on the simultaneous amplification of both genes, resulting in two distinct amplicons (947 bp and 535 bp) in heterogametic females and only a single band (535 bp) in homogametic males, which is easy to detect with agarose gel electrophoresis.4. This technique is simple and convenient for genetic sex determination in Chukar partridges.

Sex identification of ducklings based on acoustic signals.

Sex identification of ducklings is a critical step in the poultry farming industry, and accurate sex identification is beneficial for precise breeding and cost savings. In this study, a method for identifying the sex of ducklings based on acoustic signals was proposed. In the first step, duckling vocalizations were collected and an improved spectral subtraction method and high-pass filtering were applied to reduce the influence of noise. Then, duckling vocalizations were automatically detected by using a double-threshold endpoint detection method with 3 parameters: short-time energy (STE), short-time zero-crossing rate (ZCR), and duration (D). Following the extraction of Mel-Spectrogram features from duckling vocalizations, an improved Res2Net deep learning algorithm was used for sex classification. This algorithm was introduced with the Squeeze-and-Excitation (SE) attention mechanism and Ghost module to improve the bottleneck of Res2Net, thereby improving the model accuracy and reducing the number of parameters. The ablative experimental results showed that the introduction of the SE attention mechanism improved the model accuracy by 2.01%, while the Ghost module reduced the number of model parameters by 7.26M and the FLOPs by 0.85G. Moreover, this algorithm was compared with 5 state-of-the-art (SOTA) algorithms, and the results showed that the proposed algorithm has the best cost-effectiveness, with accuracy, recall, specificity, number of parameters, and FLOPs of 94.80, 94.92, 94.69, 18.91M, and 3.46G, respectively. After that, the vocalization detection score and the average confidence strategy were used to predict the sex of individual ducklings, and the accuracy of the proposed model reached 96.67%. In conclusion, the method proposed in this study can effectively detect the sex of ducklings and serve as a reference for automated sex identification of ducklings.

Temperature regulates sex determination and growth in the paralichthid flatfish California halibut.

California halibut (Paralichthys californicus) is a candidate species for aquaculture and stock enhancement. These applications rely on sex control, either to maximize the production of faster growing females or to match sex ratios in the wild. Other paralichthids exhibit temperature-dependent sex determination (TSD), but the presence and pattern of TSD is not well defined in California halibut. Juvenile California halibut were cultured at three distinct temperatures (15°C, 19°C, and 23°C) through the developmental period presumed to be thermosensitive based on findings from congeners. Sex ratios were quantified in each treatment using phenotypic sex identification techniques applied early (molecular biomarkers; 51-100 mm total length [TL]) and late (visual examination of the gonads; ≥100 mm TL) in the juvenile phase. Both techniques indicated similar sex determination trends at each temperature, with overall sex ratios assessed as 49.9% male at 15°C, 74.5% male at 19°C, and 98.2% male at 23°C. Growth rates were highest at 23°C and lowest at 15°C, with intrinsically fast- and slow-growing individuals at all temperatures. At 15°C and 19°C, females comprised a higher proportion among the fast growers than they did among the slow growers. These data show that California halibut exhibit TSD, with temperatures of 19°C and 23°C masculinizing fish while 15°C appears to produce a 1:1 sex ratio. This study will help optimize sex ratios and growth in hatcheries through thermal manipulation. Furthermore, the developed biomolecular tools and identified temperature thresholds will be important in future work to understand the influence of global warming on wild population demographics.

Gibberellins regulate masculinization through the SpGAI-SpSTM module in dioecious spinach.

The sex of dioecious plants is mainly determined by genetic factors, but it can also be converted by environmental cues such as exogenous phytohormones. Gibberellic acids (GAs) are well-known inducers of flowering and sexual development, yet the pathway of gibberellin-induced sex conversion in dioecious spinach (Spinacia oleracea L.) remains elusive. Based on sex detection before and after GA3 application using T11A and SSR19 molecular markers, we confirmed and elevated the masculinization effect of GA on a single female plant through exogenous applications of GA3, showing complete conversion and functional stamens. Silencing of GIBBERELLIC ACID INSENSITIVE (SpGAI), a single DELLA family protein that is a central GA signaling repressor, results in similar masculinization. We also show that SpGAI can physically interact with the spinach KNOX transcription factor SHOOT MERISTEMLESS (SpSTM), which is a homolog of the flower meristem identity regulator STM in Arabidopsis. The silencing of SpSTM also masculinized female flowers in spinach. Furthermore, SpSTM could directly bind the intron of SpPI to repress SpPI expression in developing female flowers. Overall, our results suggest that GA induces a female masculinization process through the SpGAI-SpSTM-SpPI regulatory module in spinach. These insights may help to clarify the molecular mechanism underlying the sex conversion system in dioecious plants while also elucidating the physiological basis for the generation of unisexual flowers so as to establish dioecy in plants.

Molecular sexing of birds using quantitative PCR (qPCR) of sex-linked genes and logistic regression models.

The ability to sex individuals is an important component of many behavioural and ecological investigations and provides information for demographic models used in conservation and species management. However, many birds are difficult to sex using morphological characters or traditional molecular sexing methods. In this study, we developed probabilistic models for sexing birds using quantitative PCR (qPCR) data. First, we quantified distributions of gene copy numbers at a set of six sex-linked genes, including the sex-determining gene DMRT1, for individuals across 17 species and seven orders of birds (n = 150). Using these data, we built predictive logistic models for sex identification and tested their performance with independent samples from 51 species and 13 orders (n = 209). Models using the two loci most highly correlated with sex had greater accuracy than models using the full set of sex-linked loci, across all taxonomic levels of analysis. Sex identification was highly accurate when individuals to be assigned were of species used in model building. Our analytical approach was widely applicable across diverse neognath bird lineages spanning millions of years of evolutionary divergence. Unlike previous methods, our probabilistic framework incorporates uncertainty around qPCR measurements as well as biological variation within species into decision-making rules. We anticipate that this method will be useful for sexing birds, including those of high conservation concern and/or subsistence value, that have proven difficult to sex using traditional approaches. Additionally, the general analytical framework presented in this paper may also be applicable to other organisms with sex chromosomes.

The development rule of feathers and application of hair root tissue in sex identification of Yangzhou geese.

Accurate gender identification is crucial for the study of bird reproduction and evolution. The current study aimed to explore and evaluate the effectiveness of a noninvasive method for gender identification in Yangzhou geese. In this experiment, 600 goose eggs were collected. Hair root tissues were used for PCR amplification, molecular sequencing, and anal inversion for early sex recognition in goslings. According to the DNA amplification results for the feather pulp tissue of 2-wk-old geese, bands appeared at 436 bp (CHD1-Z) and 330 bp (CHD1-W) upon gel electrophoresis. This method considered the base of goose feathers to accelerate the process of gender recognition. By examining the sex of anatomized poultry for verification, the accuracy rate of PCR gel electrophoresis and molecular sequencing sex identification was 100%, whereas the average accuracy rate of anal inversion was 97.41%. In the comparison of feather growth trends at 0 to 18 wk of age, the feather root weight (FRW), feather root length (FRL), feather branch length (FBL), and feather shaft diameter (FSD) of Yangzhou goose of the same age were not significantly different between males and females (P > 0.05). At 6 wk of age, the FRW, FRL, and FSD in males and FRL in females increased rapidly; their growth increased by 84.43, 67.58, 45.10, and 69.42%, respectively. At 10 wk of age, the male FRL, male FBL, and female FBL increased by 37.31, 34.81, and 21.72, respectively. The Boltzmann model was found to be the best-fitting model for the feathers of male Yangzhou geese. Early sex identification based on feather growth trends between the sexes is not feasible. This study provides a convenient and reliable technical means for early sex identification of waterfowl and serves as an ecological strategy for protecting the reproduction of poultry populations.

Morphometric variations and nonmetric anatomical traits or anomalies of the primary molar teeth, plus the molars' size thresholds for sex identification.

INTRODUCTION: Morphological and morphometric features of the teeth are of interest to various clinical and academic dental and medical fields including prosthodontics, orthodontics, anatomy and anthropology, pathology, archeology, and forensic dentistry. These have been more or less researched in the case of the permanent dentition. However when it comes to the primary dentition, the literature is scarce and controversial. No study worldwide exists on the cutoff points (thresholds) for sex identification; no study exists on metric or nonmetric traits of deciduous teeth in Iranians. Hence, the aim of the study was to assess both the metric and nonmetric traits of primary molars, as well as their cut-off points for sex identification. METHODS: In this epidemiological cross-sectional study, pretreatment casts of 110 children (51 boys and 59 girls) aged 6 to 12 years were collected. Maxillary and mandibular first and second primary molars were evaluated regarding their metric traits (mesiodistal and buccolingual widths) and 9 nonmetric traits (Accessory cusp on the upper D, Accessory cusp on the lower D, Fifth cusp on the upper E, Carabelli's cusp on the upper E, Protostylid on the lower E, Fifth cusp on the lower E, Sixth cusp on the lower E, Tuberculum intermedium [metaconulid] on the lower E, and Deflecting wrinkle on the lower E). ROC curves were used to identify cut-off points for sex determination as well as the usefulness of metric measurements for this purpose. Data were analyzed using independent-samples and paired-samples t-tests, McNemar, Fisher, and chi-square tests, plus Pearson and Spearman correlation coefficients (α = 0.05). RESULTS: All the primary molars' coronal dimensions (both mesiodistal and buccolingual) were extremely useful for sex identification (ROC curves, all P values ≤ 0.0000099). Especially, the mandibular primary molars (areas under ROC curves [AUCs] between 85.6 and 90.4%, P values ≤ 0.0000006) were more useful than the maxillary ones (AUCs between 80.4 and 83.1%, P values ≤ 0. 0000099). In the mandible, the first primary molar (maximum AUC = 90.4%) was better than the second molar (maximum AUC = 86.0%). The optimum thresholds for sex determination were reported. Sex dimorphism was significant in buccolingual and mesiodistal crown widths of all the primary molars (all P values ≤ 0.000132), but it was seen only in the case of 2 nonmetric traits: Deflecting wrinkle (P = 0.001) and Tuberculum intermedium (metaconulid, P = 0.029) on the lower Es, taking into account the unilateral and bilateral cases. The occurrence of nonmetric traits was symmetrical between the right and left sides (all P values ≥ 0.250). All mesiodistal and two buccolingual molar measurements were as well symmetrical (P > 0.1); however, two buccolingual measurements were asymmetrical: in the case of the maxillary E (P = 0.0002) and mandibular D (P = 0.019). There were three weak-to-moderate correlations between the nonmetric traits of the mandibular second molars (Spearman correlations between 22.7 and 37.5%, P values ≤ 0.045). Up to 6 concurrent nonmetric traits were observed in the sample, with 53.6% of the sample showing at least 2 concurrent nonmetric traits at the same time, without any sex dimorphism (P = 0.658). CONCLUSION: Sex dimorphism exists considerably in primary molars' sizes, but it is not as prevalent in their nonmetric traits or abnormalities. Primary molars' crown sizes are useful for sex identification; we calculated optimum cut-off points for this purpose, for the first time.

An evaluation of DNA sample source and molecular markers to determine gender in the short-beaked echidna (Tachyglossus aculeatus).

The short-beaked echidna is sexually monomorphic such that gender identification without veterinary intervention is challenging. The aim of this study was to evaluate and compare the most optimal noninvasive genetic source by extracting echidna genomic DNA (gDNA) from fecal scats, plucked hair, and quills to perform genetic sex testing using a range of molecular markers. Sex determination of 14 captive short-beaked echidnas was determined by amplifying isolated DNA from noninvasive samples, targeting two Y chromosome (male-specific) genes (mediator complex subunit 26 Y-gametologue [CRSPY] and anti-Müllerian hormone Y-gametologue [AMHY]), in addition to four confirmed sex-specific RADseq markers. Results of noninvasive samples were compared with blood samples and clinical records. Receiver operating characteristic curves were used to assess accuracy of sex determination of markers for each sample type. The gender of the echidnas was successfully identified on 75% of occasions using fecal samples, 90.6% occasions using hair, and 84.6% occasions with quills. Overall, the male-specific RADseq markers accurately identified the sex of echidnas with all sample types for 90% of animals; compared with 81.5% using CRSPY, and 82.0% using AMHY to identify sex. Collection of hair, quills, and feces provides a useful alternative to invasively collected samples, however, the accuracy of results depends on sample type and genetic marker selected. We found gender determination in the short-beaked echidna was most accurate using four male-specific RADseq markers on gDNA isolated from blood and hair. The noninvasive genetic sexing techniques documented here will inform and facilitate husbandry and genetic management of captive echidna populations.

Integrated Analysis of Machine Learning and Deep Learning in Silkworm Pupae (Bombyx mori) Species and Sex Identification.

Hybrid pairing of the corresponding silkworm species is a pivotal link in sericulture, ensuring egg quality and directly influencing silk quantity and quality. Considering the potential of image recognition and the impact of varying pupal postures, this study used machine learning and deep learning for global modeling to identify pupae species and sex separately or simultaneously. The performance of traditional feature-based approaches, deep learning feature-based approaches, and their fusion approaches were compared. First, 3600 images of the back, abdomen, and side postures of 5 species of male and female pupae were captured. Next, six traditional descriptors, including the histogram of oriented gradients (HOG), and six deep learning descriptors, including ConvNeXt-S, were utilized to extract significant species and sex features. Finally, classification models were constructed using the multilayer perceptron (MLP), support vector machine, and random forest. The results indicate that the {HOG + ConvNeXt-S + MLP} model excelled, achieving 99.09% accuracy for separate species and sex recognition and 98.40% for simultaneous recognition, with precision-recall and receiver operating characteristic curves ranging from 0.984 to 1.0 and 0.996 to 1.0, respectively. In conclusion, it can capture subtle distinctions between pupal species and sexes and shows promise for extensive application in sericulture.

Machine Learning Models for Prediction of Sex Based on Lumbar Vertebral Morphometry.

BACKGROUND: Identifying skeletal remains has been and will remain a challenge for forensic experts and forensic anthropologists, especially in disasters with multiple victims or skeletal remains in an advanced stage of decomposition. This study examined the performance of two machine learning (ML) algorithms in predicting the person's sex based only on the morphometry of L1-L5 lumbar vertebrae collected recently from Romanian individuals. The purpose of the present study was to assess whether by using the machine learning (ML) techniques one can obtain a reliable prediction of sex in forensic identification based only on the parameters obtained from the metric analysis of the lumbar spine. METHOD: This paper built and tuned predictive models with two of the most popular techniques for classification, RF (random forest) and XGB (xgboost). Both series of models used cross-validation and a grid search to find the best combination of hyper-parameters. The best models were selected based on the ROC_AUC (area under curve) metric. RESULTS: The L1-L5 lumbar vertebrae exhibit sexual dimorphism and can be used as predictors in sex prediction. Out of the eight significant predictors for sex, six were found to be particularly important for the RF model, while only three were determined to be important by the XGB model. CONCLUSIONS: Even if the data set was small (149 observations), both RF and XGB techniques reliably predicted a person's sex based only on the L1-L5 measurements. This can prove valuable, especially when only skeletal remains are available. With minor adjustments, the presented ML setup can be transformed into an interactive web service, freely accessible to forensic anthropologists, in which, after entering the L1-L5 measurements of a body/cadaver, they can predict the person's sex.

Origin of purple asparagus cultivar 'Pacific Purple' based on the sequence of sex determination gene.

Garden asparagus is one of the most important crops worldwide. Since this crop is dioecious and male plants generally have higher yields compared to female plants, several DNA markers for sex identification have been developed for acceleration of asparagus breeding. Among these markers, Asp1-T7sp and MSSTS710 were found to be effective in sex determination for many asparagus cultivars. However, we previously found that these markers were not completely suitable for sex identification in the purple asparagus cultivar 'Pacific Purple'. There are two types of male individuals in this cultivar: One type is PP-m, which is identified the sex type by Asp1-T7sp and MSSTS710 markers, while the other type is PP-m* whose sex type is not identified by these markers. Since the sex identification markers are located on the non-recombining Y region, it was expected that the sequence around this region might be different between PP-m and PP-m*. In this study, the sequence of one of the sex-determining genes, MSE1/AoMYB35/AspTDF1, was analyzed, and a comparative analysis was conducted among PP-m and PP-m* of 'Pacific Purple', A. officinalis and related species A. maritimus. The results revealed that PP-m and PP-m* has the similar sequence of MSE1/AoMYB35/AspTDF1 gene from A. officinalis and A. maritimus, respectively. 'Pacific Purple' is a cultivar developed through polycross hybrid from Italian landrace 'Violetto d'Albenga' (VA), suggesting that VA originated from an interspecific crossing between A. officinalis and A. maritimus and that the pollen parent used in 'Pacific Purple' breeding contained two types of male individuals with different MSE1/AoMYB35/AspTDF1 sequence. As a result, PP-m and PP-m* of 'Pacific Purple' harbors the similar sequences of the MSE1/AoMYB35/AspTDF1 gene from A. officinalis and A. maritimus, respectively.

Canine Crown Sexual Dimorphism in a Sample of the Modern Croatian Population.

Sex assessment is a key part of forensic analysis to establish the identity of unknown deceased individuals. Previous studies have shown that canines are the most dimorphic teeth, but population-specific data are necessary for forensic methods. This study explores sex dimorphism in canine crown dimensions and morphology in a contemporary Croatian population. The material consisted of 302 dental casts (147 females, 155 males) of orthodontic patients and dental students (11-25 years). The distal accessory ridge (DAR) of the upper and lower canines was evaluated using the Arizona State University Dental Anthropology System. Mesiodistal (MD) and buccolingual (BL) crown dimensions were measured on 120 casts. Sex differences in MD and BL dimensions were significant (p < 0.05) for all the canines (upper and lower, left and right), while in DAR only for lower canines (p < 0.000001). When all variables were put into the model, backward stepwise discriminant function analysis isolated lower canine DAR and lower left canine MD as the two independent variables differentiating sex. Using these two variables, a discriminant function formula allowed for sex determination with an accuracy of 73.5%. This study shows that both canine crown morphology and dimensions are useful for sex determination, especially for lower canines. These methods can be applied to children, as lower canines erupt at about 9 years of age.

SNP Panel and Genomic Sex Identification in Atlantic Halibut (Hippoglossus hippoglossus).

The ability to identify sex is necessary in population biology for a proper understanding of the dynamics of a population. In Atlantic halibut, phenotypic sex identification is not possible due to the lack of significant external morphological differences. We developed an Illumina SNP panel for Atlantic halibut with 4000 SNPs spread evenly throughout the genome with a minor allele frequency MAF ≥ 0.4, except for N = 249 SNPs located in a sex-determining region on chromosome 12, N = 176 of these SNPs were selected to genetically identify male and female individuals using a DAPC analysis. The genomic identification of sex allows for non-lethal sex determination and validation of sex identification in the field. The SNP panel is a new genomic resource for Atlantic halibut that will make it possible to generate the genotypic data for the large number of individuals needed to estimate population abundance using genomics and the Close Kin Mark Recapture (CKMR) approach, an emerging component of fisheries management and stock monitoring.