Beloniformes is an order composed of six families of marine and freshwater fish with elongated bodies. They are commonly known as needlefishes or long toms. In this study, the mtDNA of 32 species was examined. The four putative clusters can be seen in Figure 6A. The Silhouette plot shows an optimum of four clusters, but this may be due to distortion in the data. The Hopkins clustering value is 0.777, which represents good clustering. All four clusters are statistically significant. The results of the analysis of these 32 species are available in Supplementary File 3. Besides the three control species, three putative baramins were found. These include twelve species of exclusively marine fish, mainly from the genus Cheilopogon, but also Cypselurus hiraii, Exocoetus volitans, Hirundichthys rondeletii, and Prognichthys sealei. These are all species from the family Exocoetidae or flying fishes. This group has a normalized water type entropy value of 0, with all species being oceanodromous; fully adapted to saltwater. Besides this, ten species from various genera (Ablennes, Cololabis, Strongylura) mainly from Belonidae also formed a cluster. However, several species from other families also are part of this cluster, such as Dermogenys pusilla, five species from Hyporhamphus, and Parexocoetus brachypterus. These fish come from a mix of freshwater, brackish water, and saltwater environments, and have a normalized water-type entropy value of 0.958 (see Figure 6C). Four species were annotated as oceanodromous in FishBase, two as anadromous, and one as potamodromous. Also, ten species of Oryzias (ricefish) were found that populate mainly freshwater and brackish water. The normalized water-type entropy value is 0.579. Of the six Oryzias species that had migratory annotation from FishBase, five were non-migratory, and only one was amphidromous. This indicates that these fish species have fairly well adapted to environments with lower salinity levels. 4. Characiformes This order contains around 2,000 species classified into 18 families. Fish such as characins, piranhas, and tetras belong to this group. A total of 54 species from Characiformes were studied. According to Figure 7A, nine putative holobaramins were found, with a Hopkins clustering value of 0.778, indicating good clustering. The Silhouette plot in Figure 7B also shows an optimum of nine clusters. A list of species clusters and statistics is available in Supplementary File 4. However, cluster #8, made up of three species, has a statistically insignificant p-value of 0.148. A characteristic of these Characiformes clusters is that almost all species live in freshwater, meaning that they have almost completely adapted to this type of environment. With the exception of cluster #1, all other groups had a normalized entropy value of 0. All ten characids with migratory FishBase annotation were potamodromous. 5. Clupeiformes This diverse group includes species such as anchovies and herrings and other fish that are caught for human consumption. Its 400 species populate marine, euryhaline and freshwater environments in tropical, subtropical, and temperate climate zones (Lavoué et al. 2014). Mitochondrial genomes from 96 species from Clupeiformes were analyzed. The results can be seen in Figure 8, and are available in Supplementary File 5. The Silhouette plot in Figure 8B shows eight putative clusters (the outlier group and seven clupeiform baramins). The Hopkins clustering statistic is 0.832, which indicates good clustering. However, cluster #4, containing 17 species, has a statistically insignificant p-value of 0.342. Lavoué et al. (2013) also analyzed the mitogenomes of Clupeiformes and uncovered nine main lineages. In the present analysis, of the 93 species that were also studied by Lavoué et al., 75 (80.6%) fell into the same lineage as defined by Lavoué et al. Cluster #5 of the present analysis of Clupeiformes corresponds to lineage 3+Pristigasteridae, and clusters #6 and #7 of the present study both correspond to the family Engraulidae as defined by Lavoué et al. However, Engraulidae is made up of the subfamilies Engraulinae and Coiliinae, thus reflecting the clustering discovered in this analysis. Of the five Coiliinae species, all five can live in freshwater and brackish water, with only three species inhabiting saltwater. The species from the subfamily Engraulinae, on the other hand, inhabit mainly brackish water and saltwater, with 15 and 17 species, respectively. Only seven engraulids inhabit freshwater. Thus, since adaptation to freshwater and saltwater are occurring in opposite trajectories for these two groups, it may be that these two groups are separate holobaramins. The normalized water type entropy values are all above 0.85 indicating that all the clusters were euryhaline (adaptable to different water salinities). Since all these clusters are euryhaline, this seems to indicate that not much time has elapsed since the Flood, after which fish species would have had the chance to adapt to environments with narrower salinities. For example, Wilson et al. (2008) found that the freshwater herring (Clupea) species of Lake Tanganyika could be the result of a recent invasion by their marine relatives, which have not diverged much from their freshwater counterparts in their morphology. When the waters of the Flood receded, some portions could have formed inland lakes in Africa, whereas the rest drained off the continent. These specific Clupea species could have been localized to that portion of the receding waters that ended up inland, and hence adapted to freshwater circumstances, whereas their baraminic relatives adapted to a saltwater environment in the ocean, being localized to the portion of the receding Flood waters that drained into the ocean. 6. Cyprinodontiformes Cyprinodontiformes include small-sized fish, such as killifishes, minnows, pupfishes, and livebearers, which live mainly in freshwater and brackish water. They are represented by 1,400 species. The mtDNA of 69 species was analyzed. The results can be seen in Figure 9, and are also available in Supplementary File 6. The Hopkins clustering statistic is 0.831, which means good clustering. Besides the outlier group, there are eight putative holobaramins as indicated by the Silhouette plot in Figure 9B. The first seven groups are statistically significant. Five of the eight groups contain no saltwater species (see Figure 9C), and there are only three species, one from each of the remaining three groups, which can live in saltwater. The majority of the species (56 out of 69) are non-migratory, which means that these species have well adapted to freshwater environments (see Figure 9C). The three species that can live in saltwater, Cyprinodon variegatus, FunCSERHATI Molecular baraminology of fish 2023 ICC 190
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