Genetic relationships between and within some Malawian cichlid genera.

Genetic relationships were examined for a total of 90 individuals of 90 species or species var. from 13 genera of Malawian cichlids based on the sequences of an amplified 991-bp fragment of the mtDNA control region (mtDNA-CR). In the network analysis, no exclusive clades were made by all the members of any genera in this study. However, congeneric clades were observed by genera Buccochromis, Copadichromis, Protomelas, and Sciaenochromis, whereas no congeneric clades were observed by genera Mylochromis, Nimbochromis, and Otopharynx. In non-mbuna, an Aulonocara-Lethrinops group was divided into two groups, and the mean genetic distance of the larger group was much lower from mbuna than from other non-mbuna. Overall mean genetic distance within a genus was generally low in mbuna, whereas it was relatively high in non-mbuna. In the genetic tree of each genus, two or more large clades were observed, and some clades, such as those of Aulonocara hansbaenschi and Aulonocara nyassae in genus Aulonocara, Lethrinops micrentodon and Lethrinops sp. "gold harbor" in Lethrinops, and Otopharynx ovatus and Otopharynx brooksi in Otopharynx, were very deeply differentiated. Besides, a mbuna species, Pseudotropheus crabro, was extremely deeply differentiated from other members of this genus. These results suggest a widespread morphological convergence across the taxa in parallel with deep genetic differentiation in the long evolutionary story and some possibility of generation of the species of Aulonocara-Lethrinops group by hybridization of small non-mbuna and mbuna species. Furthermore, taxonomical reexamination is necessary based on a strong support by genetic connection.

Immunomodulation and Humoral Immune Response in Teleost Immunized with Aeromonas-Derived Antigenic Extracellular Bioactive Molecules.

The common use of antimicrobials in food-animal production can lead to drug residues in edible tissues for consumers. However, immunomodulators enhance immune responses and vaccine effectiveness. A new perspective explores bacterial extracellular bioactive molecules (EBMs) in food-animal production to modulate host immune responses, potentially transforming pathogen management and antimicrobial use. This study investigates the immunogenic potential of Aeromonas hydrophila-derived EBMs (Antigens) to enhance the immune system. Four Antigens were administered intraperitoneally to Oreochromis niloticus (Nile Tilapia). Antigens 2 and Antigens 3 boosted fish immune competence within 21 days. Remarkably, Antigens 3 induced robust immunity against A. hydrophila with a single dose, notably enhancing antibody-based immune responses. The increased antibody activity suggests Antigens 3 could be a vaccine candidate, promising further research and potential application in food-animal production to improve disease control. This study highlights immunomodulators' potential in reshaping disease management in the food-animal industry, emphasizing the benefits of focusing on bacterial EBMs to reduce reliance on antimicrobials and achieve sustainable disease prevention.

Molecular phylogeny of Japanese marine Tanytarsini chironomids (Chironomidae: Chironominae).

Tanytarsini is a large tribe of Chironomidae with at least 11 recorded marine species grouped in three genera. In this study, we performed a phylogenic analysis using molecular data from 13 Tanytarsini genera, including all Japanese marine species in a large tribe of Chironominae, to estimate their phylogenetic positions and evolutionary history. The phylogenetic reconstruction grouped the marine species in two clades. One clade was composed of five marine Tanytarsus and two marine Pontomyia species within a larger clade of Tanytarsus. Pontomyia is considered to have morphologically specialized and adapted to marine environments by rapid evolution, although it formed a clade with Tanytarsus. The only one species of Tanytarsus, T. pelagicus, clustered as a member of the second clade, which was mainly composed by species of the genus Paratanytarsus. Thus, we redescribe T. pelagicus as Paratanytarsus pelagicus.

Antibacterial activity of Mallotus japonicus (L.F.) Müller Argoviensis on growth of Aeromonas hydrophila, A. salmonicida, Edwardsiella tarda and Vibrio anguillarum.

AIMS: The present study evaluated the antimicrobial activities of the medicinal plant Mallotus japonicus against the fish pathogenic bacteria, Aeromonas hydrophila, Aeromonas salmonicida, Edwardisella tarda and Vibrio anguillarum, and also describes the antimicrobial activities of the major and minor active compounds present within the plant extract. The synergistic effects by way of combination of these compounds were also evaluated and described. Chemical constituents of the plant extracts were analysed using the liquid chromatography-mass spectrometry (LC-MS) and described. METHODS AND RESULTS: The diethyl ether-extract of the plant elicited the strongest antibacterial activity against the challenged bacterial species, followed by ethanol- and methanol-extracts. The major active compound of the extracts, bergenin, demonstrated no antibacterial activity, but other compounds in the extracts did. CONCLUSION: Mallotus japonicus could be used as a prophylaxis to treat bacterial disease infections of fishes and its diethyl ether-extract has the potential of an alternative to antibiotic treatment in aquaculture. SIGNIFICANCE AND IMPACT OF THE STUDY: Mallotus japonicus diethyl ether-extract has the potential of an alternative to antibiotic treatment in aquaculture.

16S rRNA Gene Sequence Identification of Cultivable-Bacterioplankton Between Ambient Water and Gastrointestinal Tract (GIT) of Resident Teleost.

This study was conducted to evaluate the ambient water bacterial dynamics and its influence on the gut of resident teleosts; Oreochromis niloticus. The bacterial communities in the ambient water and the gastrointestinal tract (GIT) of O. niloticus were profiled using a culture-dependent method and followed by the 16S rRNA gene sequencing. The results indicated bacterial phyla of Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes respectively between the two microbial consortia. However, the relative abundance among the bacterial phyla notably differed between the two consortia. The relative abundance of Proteobacteria (≤ 67%) was dominant in both consortia, but contrarily, Actinobacteria and Bacteroidetes were higher in the ambient water relative to the GIT which indicated Proteobacteria and Bacilli as the highest diversity. Nevertheless, the relative abundance of γ-proteobacteria and Bacilli remarkably dominated both consortia at the class level, with Bacillus and Pseudomonas being the most abundant operational taxonomic units (OTUs).

Host range differences between two species of freshwater horsehair worm (Nematomorpha: Chordiidae) Chordodes japonensis and C. formosanus in Japan.

Horsehair worms (Nematomorpha: Gordioidea) are endoparasites of terrestrial arthropods, of which two species, Chordodes japonensis and C. formosanus, may have been historically confused due to their morphological similarity. In this study, we conducted field surveys and laboratory parasite inoculation experiments to clarify the characteristics and differences in host range and specificity between the two species. The field surveys revealed that the host ranges of the two species are markedly different. C. formosanus parasitized diverse mantids, while C. japonensis infected only species in the genus Tenodera, as predicted in previous studies. However, the two species had one mantid host, T. angustipennis in common. Parasite inoculation experiments using three species of mantids, revealed parasitism in each of the host-parasite combinations same as in the field study. These results suggest that the observed differences in host patterns are due to physiological factors between the host and parasite. Furthermore, cross-testing conducted on the shared host T. angustipennis showed that the two species coexisted within the same host in some cases, suggesting that the competitive exclusivity of the two species is low. This study also presents a tentative list of host ranges for the two species, comparing and integrating information from this and previous studies. However, the possibility remains that the host ranges of both species may be more diverse.

Species-specific Microorganisms in Acid-tolerant Chironomus Larvae Reared in a Neutral pH Range under Laboratory Conditions: Single Dataset Analysis.

To obtain a more detailed understanding of organismal acid tolerance, the larval microbiomes of 11 Chironomus species collected from acidic or neutral pH areas in Japan and reared at pH 7-8 under laboratory conditions were systematically compared using an amplicon sequencing ana-lysis. Evenness values were lower for the larval microbiomes of acid-tolerant Chironomus cf. riparius, Chironomus fusciceps, and Chironomus sulfurosus than for eight acid-sensitive species based on an alpha diversity ana-lysis. The lower evenness observed suggested a biased abundance of microorganisms, which was consistent with the identification of Chironomus species-specific microorganisms (such as Agromyces mediolanus and Comamonas odontotermitis related bacteria) with high abundance in acid-tolerant larvae. The abundance of specific microorganisms was also high in the microbiome of acid-tolerant larvae of Chironomus acerbiphilus reared at pH 4, but not in that of acid-sensitive larvae. Based on a PICRUSt2 ana-lysis, genes involved in saccharide transport were less abundant in the microbiome of acid-tolerant larvae than in that of acid-sensitive larvae, indicating nutrient-poor acidic environments. Although these results were obtained from single datasets, acid-tolerant larvae appeared to establish Chironomus species-specific interactions with microorganisms independent of saccharides, in contrast to acid-sensitive larvae. The present study is the first step towards understanding organismal acid tolerance.

Transcriptome analysis of Chironomus sulfurosus larvae living in acidic environments: Insights into molecular mechanisms for acid tolerance.

Larvae of chironomid Chironomus sulfurosus mainly live in acidic rivers near hot springs, suggesting that they naturally select acidic environments as preferred habitats. Here we showed that C. sulfurosus larvae moved toward acidic areas and stayed alive on agar gels with a pH gradient of H2SO4, and the body fluid pH of the homogenized larvae was near neutral even acclimated under the acidic conditions, indicating mechanisms for acid tolerance. In order to gain insights into this mechanism at the molecular level, de novo RNA-seq analysis was performed on C. sulfurosus larvae. As a result, 1,208 genes were found to be significantly up-regulated in larvae acclimated at pH 2.0 compared to controls at pH 7.0. Among the up-regulated genes, ones encoding cuticle proteins, peritrophic matrix proteins, mucus-forming proteins, F-type ATPase subunits, glutathione S transferases, ß-1,3-D-glucan synthetase, hemoglobin, and cytochrome P450 were identified. This transcriptome analysis in conjunction with behavioral and biochemical assays expands our knowledge of gene expression in C. sulfurosus larvae living in acidic environments, which will provide a basis for further studies to elucidate the molecular mechanisms for acid tolerance employed by organisms in nature.

Temporal and spatial dynamics of a lower-intertidal lancelet population in the Seto Inland Sea, Japan.

The population dynamics of Branchiostoma japonicum, formerly known as B. belcheri, were Investigated from September 2003 to August 2005 in the intertidal zone at Takehara (Hiroshima Prefecture), Seto Inland Sea, Japan. The intertidal population appeared from spring to autumn and disappeared during winter. A laboratory experiment showed that exposure to temperatures below 1 degree C for 2 hours resulted in severe mortality. This low temperature corresponds to the minimum temperature in sediments in the study area. This result suggested either that the intertidal population collapses in winter because of low temperature, or that the lancelets escape from the intertidal to the subtidal zone. Throughout the research period, no lancelets smaller than 10 mm in body length were found, indicating that no larvae settled in the intertidal zone. The intertidal population is probably maintained by the influx of individuals from the neighboring subtidal population. The mean annual density of the lancelets was greatest (10.6 individuals/m(2)) at station 1 nearest the low water mark, and lowest (0.3 Individuals/m(2)) at station 3 furthest from the low water mark. In summer, the water content of the sediments was remarkably lower at station 3 (20.2%) than at station 1 (25.8%). Another laboratory experiment showed that higher mortality occurred from exposure to sediments with a water content less than 25% for 2 hours, comparable to the water content at station 3, suggesting that the spatial distribution of the lancelets upward in the intertidal is restricted by sediment dryness.

A new species of Tanytarsus van der Wulp, 1874 (Diptera: Chironomidae) and the first Japanese record of Tanytarsus ovatus Johannsen, 1932 with DNA barcodes.

A new species of the genus Tanytarsus van der Wulp, 1874 was described and illustrated based on an adult male. Tanytarsus trichovalis sp. nov. belongs to the eminulus species group, and is closely related to T. tamaundecimus Sasa, 1980, T. okuboi Sasa, 1986, and T. tonebeceus Sasa Tanaka, 2000. However, the species is distinguished from these species by the morphology of the hypopygium. We also present the first record of T. ovatus Johannsen, 1932 in Japan. In addition, DNA barcoding of 5 species including T. trichovalis sp. nov., T. ovatus, and other morphologically related species was undertaken. A new key of the eleven Japanese eminulus species group was compiled.

Renewal of genetic composition of a lancelet, Branchiostoma belcheri, in the Seto Inland Sea, Japan.

We studied renewal of genetic composition of a lancelet, Branchiostoma belcheri, at a station in the Seto Inland Sea, Japan, using random amplified polymorphic DNA (RAPD) technique. We detected a total of 49 DNA bands from 27 individuals of 5 seasonal samples. Among these, some were specific to individual or seasonal sample. There were no bands common to all individuals. We calculated the Band Sharing Index (BSI) values for each combination of individuals. BSI was in a wide range of 0.00-0.97 and the average BSI between the individuals of a sample was remarkably different from sample to sample (0.37-0.83). In cluster analysis on the basis of BSI, largely 3 clusters were constructed, which finally connected to a single cluster at as low as about 0.2 of BSI. Besides, many clusters were constructed exclusively by a seasonal sample. These results suggest a large gene pool of lancelets in this area, the remarkable differences in gene pool size among the samples and a seasonal renewal of genetic composition at a habitat.

Variations in body size of the lancelet Branchiostoma belcheri at different depths in the Seto Inland Sea: effect of food supply on the growth rate.

Variations in body size of the suspension-feeding lancelet Branchiostoma belcheri were examined from April 2001 to December 2001 at different water depths of 10-80 m. The lancelets were abundantly collected (220 indiv./dredge) at water depth of 10 m. The maximum size, ranging from 36.8 to 50.4 mm BL, decreased with increasing water depth. However, the minimum size of 7.4-7.8 mm BL, which corresponds to settling size, did not different with water depth. From seasonal changes in the length-frequency histograms, five cohorts were observed at every water depth. The lancelets grew to 44 mm BL at water depths of 10-20 m and to 34 mm BL at water depth of 80 m with four years of life span. Developmental stages of the gonads at an age of two years showed that no individuals had undeveloped gonads at water depth of 10 m, but 82% of them did at water depth of 80 m. The analysis of the stepwise multiple regressions of monthly growth rate on environmental variables showed that chlorophyll a was the best explanatory variable and showed a significant positive correlation with the growth of 1-2 years age groups. These results suggested that variations in the body length were mainly fluctuated by phytoplankton supply.