Cultivable Yeast Microbiota from the Marine Fish Species Genypterus chilensis and Seriolella violacea

Because of its outstanding biological and industrial importance, many efforts have been made to characterize the mycobiota of new environments and their biochemical and biotechnological potentials. Gut mycobiota can be a source of novel yeasts with the potential to be used as probiotics or have industrial applications. In this work, we characterized two as-yet unexplored yeast communities from the intestinal content of the cultured marine Chilean fishes Genypterus chilensis (G. chilensis) and Seriolella violacea (S. violacea). Yeasts were isolated through culture, identified by sequencing their ITS region, and characterized their enzymatic profile with API®ZYM. Rhodotorula mucilaginosa was identified in both fish species. For the first time, Candida palmioleophila, Candida pseudorugosa, Cystobasidium slooffiae, and a member of the Yamadazyma genus were also identified and described as part of the normal fish gut–microbiota. Furthermore, the diverse enzymatic profile exhibited by some of these isolates suggests that it may be possible to develop novel applications for them, such as new probiotics and other biotechnological applications.

Regulation of fish resources rational using on the territory of the NNP “BUG GARD”

Today on the territory of Ukraine there are 40 national nature parks which include the NNP “Bug Gard”. One of the main tasks of the NNP is to preserve the valuable natural objects and to develop the scientific recommendations for protecting the environment and using the natural resources efficiently. Modern poaching, with its mass, technical armament, impunity and constant rapid growth, poses a serious threat to the national security of Ukraine. Significant damage to fishing and fish farming on the territory of the NNP “Bug Gard” is caused by the actions of poachers. During the functioning of the NNP “Bug Gard” the State Protection Service constantly conducts inspections and patrols the territory of the park to identify and to stop violations of the environmental legislation. The evaluation of the effectiveness of the fish protection measures carrying out in the Bug National Nature Park by the State Protection Service has been assessed. Annually the State Protection Service of the NNP “Bug Gard” conducts on average of 120 activities to identify environmental offenses, practices the explanatory talks, on average, 600 conversations per year, which are aimed at clarifying the environmental legislation in the field of fauna and flora protection, as well as it constantly informs the public through the media about the individual cases of poaching in order to further preventing the environmental offenses. Annually, an average of 1.000 leaflets with the fire safety rules on the territory of the NNP and with the environmental issues were produced and distributed among the population and the visitors of the park. It is established that, according to the analysis, the State Protection Service of the NNP “Bug Gard” carries out the effective measures in the field of fish resources conserving and rational using in the area of its responsibility, carries out a widespread waste water pollution control in the water bodies, creates the conditions for normal fish reproduction and keeps a track of environmental and fishing offenses, which not only preserves the flora and fauna, but also compensates the damage caused by the actions of poachers and other violators.

Simple morphometrics for predicting lordosis-induced deviations of body-shape in reared Gilthead seabream (Sparus aurata L.)

Haemal lordosis, V-shape bending of the haemal vertebrae, is a frequent abnormality of reared fish. Lordosis severity ranges from light deformations of vertebral axis, with insignificant effects on external morphology, to severe axis deformations with significant impact on body-shape. In the present study, we developed a simple morphometric index (PrAn) that links lordosis severity at the juvenile stage with fish body-shape at harvesting, without requiring to radiograph or sacrifice the samples. Examined seabream specimens were part of our previous study (Fragkoulis et al. 2019, Sci. Rep. 9, 9832), which monitored the effects of lordosis on the external morphology of pit-tagged seabream juveniles during their growth, up to harvest size. At both juvenile and adult stages, PrAn was effective in discriminating the normal fish from ca the 70% of lordotic fish. Our results suggest the PrAn as a valuable scale of quality, which quantifies the lordosis effects on fish external morphology, both at the juvenile stage and at harvest. Depending on the lordosis rates, and the hatchery strategy on the maximum allowed abnormality rates, this scale can cull out different rates of lordotic fish, without affecting the fish with normal phenotype or the lordotic fish with high recovery potential.

The Contamination Effects and Toxicological Characterization of o-Chlorobenzylidene Manolonitrile

Toxicological characterization and global assessment of the toxicity of some chemicals used in the specific technique of the defense and public order system (e.g. o-chlorobenzylidene manolonitrile which enters the composition of the CS gas) is especially useful for environmental protection. Based on the results obtained, the toxicological characterization (acute lethal toxicity) and the harmful potential of some toxic substances, individual or in mixture, against euythermal sweet fish are established. The tests performed aim to change the behavior of the species studied in the presence of chlorobenzylidene manolonitrile, a substance used in the technique of defense and public order system. The tests established the toxicity values of acute lethal concentration LC50 and serve to further determine the maximum allowable concentration that inhibits the development of normal fish fauna in contaminated waters. The test results allow the toxicological characterization and measurement of the acute lethal toxicity of an inhibitor, in order to make more efficient its elimination from the contaminated environment.

A fish with no sex: gonadal and adrenal functions partition between zebrafishNR5A1co-orthologs

People with NR5A1 mutations experience testicular dysgenesis, ovotestes, or adrenal insufficiency, but we do not completely understand the origin of this phenotypic diversity. NR5A1 is expressed in gonadal soma precursor cells before expression of the sex-determining gene SRY. Many fish have two co-orthologs of NR5A1 that likely partitioned ancestral gene subfunctions between them. To explore ancestral roles of NR5A1, we knocked out nr5a1a and nr5a1b in zebrafish. Single-cell RNA-seq identified nr5a1a-expressing cells that co-expressed genes for steroid biosynthesis and the chemokine receptor Cxcl12a in 1-day postfertilization (dpf) embryos, as does the mammalian adrenal–gonadal (interrenal-gonadal) primordium. In 2dpf embryos, nr5a1a was expressed stronger in the interrenal-gonadal primordium than in the early hypothalamus but nr5a1b showed the reverse. Adult Leydig cells expressed both ohnologs and granulosa cells expressed nr5a1a stronger than nr5a1b. Mutants for nr5a1a lacked the interrenal, formed incompletely differentiated testes, had no Leydig cells, and grew far larger than normal fish. Mutants for nr5a1b formed a disorganized interrenal and their gonads completely disappeared. All homozygous mutant genotypes lacked secondary sex characteristics, including male breeding tubercles and female sex papillae, and had exceedingly low levels of estradiol, 11-ketotestosterone, and cortisol. RNA-seq showed that at 21dpf, some animals were developing as females and others were not, independent of nr5a1 genotype. By 35dpf, all mutant genotypes greatly under-expressed ovary-biased genes. Because adult nr5a1a mutants form gonads but lack an interrenal and conversely, adult nr5a1b mutants lack a gonad but have an interrenal, the adrenal, and gonadal functions of the ancestral nr5a1 gene partitioned between ohnologs after the teleost genome duplication, likely owing to reciprocal loss of ancestral tissue-specific regulatory elements. Identifying such elements could provide hints to otherwise unexplained cases of Differences in Sex Development.

Microbiota Modulates the Immunomodulatory Effects of Filifolinone on Atlantic Salmon

Filifolinone is an aromatic geranyl derivative, a natural compound isolated from Heliotropum sclerocarpum, which has immunomodulatory effects on Atlantic salmon, upregulating cytokines involved in Th1-type responses through a mechanism that remains unknown. In this work, we determined whether the immunomodulatory effects of filifolinone depend on the host microbiotic composition. We evaluated the effect of filifolinone on immune genes and intestinal microbiotic composition of normal fish and fish previously treated with bacitracin/neomycin. Filifolinone induced the early expression of IFN-α1 and TGF-β, followed by the induction of TNF-α, IL-1β, and IFN-γ. A pre-treatment with antibiotics modified this effect, mainly changing the expression of IL-1β and IFN-γ. The evaluation of microbial diversity shows that filifolinone modifies the composition of intestinal microbiota, increasing the abundance of immunostimulating organisms like yeast and firmicutes. We identified 69 operational taxonomic units (OTUs) associated with filifolinone-induced IFN-γ. Our results indicate that filifolinone stimulates the immune system in two ways, one dependent on fish microbiota and the other not. To our knowledge, this is the first report of microbiota-dependent immunostimulation in Salmonids.

REKAYASA KOMPOSISI PAKAN PELLET BENIH IKAN PATIN (Pangasianodon hypophtalmus) DENGAN FERMENTASI EM4 (Effective Microorganisms 4)

The high level feed of consumption of catfish (Pangasianodon hypopthalmus) is due to its relatively fast growth. Efforts to overcome the need for large and high quality feeds are needed to improve feed engineering through the administration of Effective Microorganisms 4 (EM4) bacteria in the hope that the protein in the pellet can increase and the texture is improved so that fish digestibility increases. The study design used was a complete randomized design (CRD) 4 (four) treatments 3 (three) replications with ANOVA analysis. Catfish weighing 5 ± 0.3 grams / head with a length of 8 ± 0.5 cm are fed commercially fermented pellet 0.5 ml EM4 / 50 ml water. 1.0 ml and 1.5 ml per kilogram of feed with protein values P0 35.1282%, P1 32.8506%, P2 27.5527%, and P3 32.3208%. The experimental results showed that absolute growth were respectively P0 2.1 grams, P1 2.3 grams, P2 1.9 grams and P3 2.1 grams and absolute length P0 1.8 cm, P1 1.8 cm, P2 1.5 cm and P3 1.7 cm. Graduation of live fish until observation day 20 amounted to 100% then entered day 30 P0 93.33% cm, P1 93.33%, P2 83.33% and P3 90.00%. Conversion of feed P0 1.3, P1 1.0 P2 1.2 and P3 1.1%. Normal fish water quality Temperature 27.0 - 27.2o C, pH 6.0-6.7, DO2 5.0-5.3 mg / l, CO2 2.1-2.5 mg / l and Ammonia 0.010-0.014 mg / l,Keywords: catfish, feed and microorganisms. Abstrak. Tingkat kosumsi pakan ikan patin (Pangasianodon hypopthalmus) yang tinggi disebabkan oleh pertumbuhannya yang relatif cepat. Upaya untuk mengatasi kebutuhan pakan yang besar dan bermutu diperlukan rekayasa perbaikan pakan melalui pemberian bakteri Effective Microorganisms 4 (EM4) dengan harapan protein pada pellet dapat menigkat dan teksturnya lebih membaik sehingga kecernaan ikan meningkat. Rancangan penelitian yang digunakan adalah rancangan acak lengkap (RAL) 4 (empat) perlakuan 3 (tiga) ulangan dengan anlisis Anova. Ikan patin ukuran bobot 5±0.3 gram/ekor dengan ukuran panjang 8±0.5 cm diberi pakan komersial pellet difermentasi 0.5 ml EM4 /50 ml air. 1.0 ml dan 1.5 ml per kilogram pakan dengan nilai protein P0 35.1282%, P1 32.8506 %, P2 27.5527%, dan P3 32.3208%. Hasil percobaan menunjukan bahwa pertumbuhan mutlak berturut-turut P0 2.1 gram, P1 2.3 gram, P2 1.9 gram dan P3 2.1 gram dan panjang mutlak P0 1.8 cm, P1 1.8 cm, P2 1.5 cm dan P3 1.7 cm. Kelulusan hidup ikan sampai pengamatan hari ke 20 berjumlah 100% kemudian memasuki hari 30 P0 93.33% cm, P1 93.33%, P2 83.33% dan P3 90.00%. Konversi pakan P0 1.3 , P1 1.0 P2 1.2 dan P3 1.1%. Kualitas air ikan peliharaan normal Suhu 27,0 – 27,2o C, pH 6.0-6.7, DO2 5,0-5,3 mg/l, CO2 2,1-2,5 mg/l dan Amoniak 0,010-0,014 mg/l,Kata Kunci : Ikan Patin, Pakan dan Microorganisme.

MON-LB63 Hypocalcemia-A Clue to Digeorge Syndrome; A Very Rare Case Diagnosed in a 41 Year Male

41-year-old male with history of anxiety, bipolar disorder, depression, cognitive developmental delay, Idiopathic Thrombocytopenic Purpura (ITP), Vit D deficiency, tinnitus, suicidal attempt, auditory hallucination, borderline intellectual functioning comes to the PCP’s office for regular follow-up. The PCP has noted that the patient was hypocalcemic on multiple visits. On exam, vitals were in normal range, height 5 feet 2 inches, BMI 31 kg/m2.Despite the patient being on calcium and Vit D2 pills for a couple of months, his Calcium level were low ranging from 5.8–7.8mg/dl (normal 8.5-10.1) with normal albumin; ionized calcium was low 0.9-0.97mmol/l (normal 1.12-1.23); Magnesium was normal 2mEq/L (normal 1.5-2.5), phosphorus slightly high 5 mg/dL (normal 2.5-4.9), PTH (ParaThyroid Hormone level) was low-normal 18.8pg/ml (normal 11.1-79.5), vitamin D 25 OH low 16ng/ml (normal 30-100); 1,25 di OH vitamin D low 8ng/ml (normal25-40).Thyroid ultrasound showed 2 benign nodules, no further work-up done. TSH and free T4 were normal. FISH (Fluorescence In Situ Hybridization) came positive for DiGeorge syndrome (deletion at 22q11.2). He was treated with calcium 600 mg 4 pills daily, calcitriol 0.25 mcg two pills daily, Vit D2 50000 IU weekly and thiazide diuretic. His labs improved. DISCUSSIONHypocalcemia can be due to low magnesium level, drugs or associated with high or low PTH. The patient had normal magnesium and he was not on any medication that causes hypocalcemia. This rules out first two casues. Hypocalcemia with high PTH (pseudohypoparathyroidism or low vit D levels) doesn’t fall in our differential because our patient had low PTH. For hypocalcemia with low PTH, differentials include post-surgical condition (no surgical history), autoimmune (history and labs not suggestive of), infiltrative diseases (eg hemochromatosis- he had normal iron study, Wilson disease- normal copper level, granulomas- contrary he had low 1,25 Di OH vit D). Hypocalcemia secondary to genetic parthyroid gland anomaly was thought to be most likely in our patient. So FISH was pursued. Conclusion: For patients with cognitive issues, persistence of chronic hypocalcemia (with low PTH) despite treatment should prompt for genetic disorders like DiGeorge. DiGeorge is usually the diagnosis of children. Perhaps this is the first case of DiGeorge diagnosed so late at age of 41.

A cryptic imatinib-sensitive G3BP1-PDGFRB rearrangement in a myeloid neoplasm with eosinophilia

Key Points Targeted RNA sequencing detected a cryptic G3BP1-PDGFRB rearrangement in a myeloid neoplasm with eosinophilia and normal FISH studies. Consistent with the patient’s response to imatinib, we demonstrate this rearrangement is oncogenic and sensitive to TKI in cell culture.

Aktifitas Selulolitik dan Patogenisitas Bacillus cereus_TSS4 dari Serasah Daun Mangrove

Leaf litter of mangrove has the potential for cellulolytic bacteria that are beneficial in aquaculture feed. Tin mining in Bangka Island impact for mangrove and allows for new strains of cellulolytic bacteria. Identification and safety evaluations are needed to knows the applied to aquaculture. This study aims to obtain and evaluate the potential impact on the aquaculture of cellulolytic bacteria from the Tukak Sadai mangrove, South Bangka. The effects were shown on the survival and clinical symptoms of fish through pathogenicity testing of the selected bacteria. This research was done from March 2017 until March 2018. Leaf litter was a sample taken from mangroves and isolated using 1% Carboxymethyl Cellulosa (CMC) media. Qualitative test of cellulase enzyme activity uses congo red and bacterial identification to use biochemical characterization with Microbact TM. There were six cellulolytic bacterial isolates from the mangrove leaves of Tukak Sadai, South Bangka. TSS4 isolates had the highest cellulolytic index of 26.4 mm compared to other strains. Biochemical characterization of TSS4 isolates show similarities with Bacillus cereus. Pathogenicity test on Bacillus cereus_TSS4 isolates show that it was not pathogenic with normal fish conditions until the end of maintenance, fish survival reached 100%, and no damage to internal organs occurred.Keywords: Bacillus cereus, mangrove leaf litter, pathogenicity, cellulolytic bacteriaAbstrak. Serasah daun mangrove memiliki potensi bakteri selulolitik yang bermanfaat pada pakan dalam akuakultur. Penambangan timah di Pulau Bangka berdampak pada hutan bakau dan memungkinkan strain bakteri selulolitik baru. Identifikasi dan evaluasi keamanan diperlukan untuk mengetahui penerapan pada budidaya. Penelitian ini bertujuan untuk mendapatkan dan mengevaluasi dampak potensial pada akuakultur dari bakteri selulolitik hutan bakau Tukak Sadai, Bangka Selatan. Dampaknya ditunjukkan pada kelangsungan hidup dan gejala klinis ikan melalui pengujian patogenisitas dari bakteri yang dipilih. Penelitian ini dilakukan mulai Maret 2017 hingga Maret 2018. Serasah daun merupakan sampel yang diambil dari mangrove dan diisolasi menggunakan media Carboxymethyl Cellulosa (CMC) 1%. Uji kualitatif aktivitas enzim selulase menggunakan congo red dan identifikasi bakteri untuk menggunakan karakterisasi biokimia dengan MicrobactTM. Ada enam isolat bakteri selulolitik dari daun mangrove Tukak Sadai, Bangka Selatan. Isolat TSS4 memiliki indeks selulolitik tertinggi 26,4 mm dibandingkan dengan jenis lainnya. Karakterisasi biokimia isolat TSS4 menunjukkan kesamaan dengan Bacillus cereus. Uji patogenisitas pada isolat Bacillus cereus_TSS4 menunjukkan bahwa tidak patogen dengan kondisi ikan normal sampai akhir pemeliharaan, kelangsungan hidup ikan mencapai 100%, dan tidak terjadi kerusakan pada organ internal.Kata kunci: Bacillus cereus, serasah daun mangrove, patogenisitas, bakteri selulolitik