Early life-history stages of the olive barb, Systomus sarana (Hamilton), and a key ontogenetic systematic character of Smiliogastrinae.

The current study aims to uncover the early life-history stages of Systomus sarana, a medium-sized smiliogastrin cyprinid important for aquaculture in South Asia. The fish were effectively bred in captivity by administering 0.25 mL of breeding hormone per kilogram of fish. The spawning occurred 8.54 ± 0.55 h after the injection, and the eggs were phyto-lithophilic with a pale yellow color and a diameter of 1.49 ± 0.04 mm. Hatching occurred 17 h after fertilization, and the yolk-sac larvae of 3.43 ± 0.08 mm total length (TL) were adhering to the plant parts and other substrata with the cement glands on the forehead. On the third day, with complete absorption of the yolk sac and the disappearance of the attachment organ, the pre-flexion larvae measured 5.3 ± 0.11 mm TL. On the eighth day, the flexion larvae measured 6 ± 0.4 mm TL with a well-inflated posterior swim bladder, and the post-flexion larvae, at 11 days post-hatching (dph), developed a two-chambered gas bladder. The juvenile stage, on day 21 post-hatching, was marked by the loss of the median finfolds and the appearance of black blotches on the caudal, subdorsal, and supra-anal regions. The commencement of squamation and the appearance of the rudiments of maxillary barbels distinguished the juvenile stage. The subadults measuring 4.6 ± 0.36 cm TL had finished squamation and completely lost the subdorsal and supra-anal blotches. We propose that the presence of subdorsal blotches is a distinctive ontogenetic and systematic feature of larval and juvenile forms of smiliogastrin barbs.

Fish assemblage and guild structure in the Ashtamudi Estuary, a tropical Ramsar site in India.

A first hand study on guild structure of fish species (n = 148) in Ashtamudi Estuary, a Ramsar Site of International importance, was carried out from May 2019 to April 2020 based on temporal (monsoon, pre-monsoon and post-monsoon seasons) and spatial (upper, middle and lower zones) data. Guild composition consisted of eight estuarine use guilds (marine estuarine opportunist, marine estuarine dependent, estuarine resident species, anadromous species, freshwater migrants, freshwater stragglers, catadromous and marine stragglers) and eight feeding guilds (zooplanktivore, zoobenthivore, carnivores, herbivores, invertebrate feeders, omnivores, detritus feeders and piscivores). The most dominant estuarine use guilds comprised of marine migrants (marine estuarine opportunist and marine estuarine dependent), whereas carnivore, omnivore and zooplanktivore formed the dominant feeding guilds in Ashtamudi Estuary. The species diversity decreased from upper to lower zone and was highest during the post-monsoon period. The present study has seen a decrease in estuarine resident and freshwater species due to reduced freshwater inflow, saltwater intrusion, unsustainable fishing and inadequate environmental management practices. Hence, we propose a management framework to improve the ecosystem of Ashtamudi Estuary with a special focus on its fish species and their estuarine use.

Treatment of hepatitis C virus infection with direct-acting antiviral agent-based regimens in Iranian patients with hereditary bleeding disorders.

BACKGROUND: Chronic hepatitis C (CHC) is one of the most important comorbidities in patients with hereditary bleeding disorders (HBD). The present study aimed at evaluating the effectiveness of direct-acting antiviral agent (DAA)-based interferon-free HCV antiviral regimens in patients with HBD. PATIENTS AND METHODS: The present study was performed on the patients with HBD and CHC between 2015 and 2019. Sofosbuvir-based interferon-free regimens with or without ribavirin were prescribed to treat HCV infection. The main endpoint of the study was to determine the sustained virologic response (SVR), assessed 12 weeks after the completion of treatment. RESULTS: A total of 147 patients with a mean age of 41.1 years were enrolled in the study; 4.1% of them were co-infected with HIV, 25.2% had cirrhosis, and 76.9% of them were diagnosed with hemophilia A. HCV genotype-1 includes the largest number (68.1%) of patients. 46.3% of patients were treatment-naïve and others had a treatment history with interferon-based regimens. Out of 147 patients, 15 patients were lost to follow-up during treatment or for SVR evaluation or discontinued treatment. 132 subjects completed treatment and were evaluated for SVR, 12 weeks after the completion of treatment. All of the patients achieved SVR 12 (SVR rate: 100%, 95% CI 97.2-100%). CONCLUSION: Hepatitis C DAA-based regimens are the effective treatments for CHC in patients with HBD, regardless of the treatment modifiers such as previous treatment experience, cirrhosis, HIV co-infection, and HCV genotype.

Outbreaks of epizootic ulcerative syndrome in Kerala, India following episodes of flooding.

Large-scale fish mortality was observed in flood-affected fish farms across several parts of Kerala following heavy rainfall in August 2018 and 2019-nearly 53% above the normal monsoon rain that the region receives. The affected fish had severe haemorrhages and ulcers, typical of the highly infectious disease epizootic ulcerative syndrome (EUS) caused by the water mould Aphanomyces invadans. In freshwater, snakeheads Channa spp. and in brackish water mullet (Mugilidae) and pearl spot (Etroplus suratensis) were severely affected. EUS was observed in 4 freshwater fishes for the first time: dotted sawfin barb Pethia punctata (Cyprinidae), Malabar leaffish Pristolepis malabarica (Pristolepididae), mahecola barb Puntius mahecola (Cyprinidae) and giant snakehead Channa pseudomarulius (Channidae). Histology and molecular diagnosis confirmed the cause of mortality to be EUS. Fungal hyphae were also observed in deeply ulcerated fish, revealed by lactophenol cotton blue staining. The severity of the EUS outbreak was linked to the sudden change in water quality associated with the flood, such as lower water temperature, and decreases in pH, total alkalinity and total hardness.

Acute changes in cardiac function by direct acting antiviral therapy for hepatitis C-infected patients with thalassemia.

BACKGROUND: Patients with thalassemia may also have cardiac abnormalities due to congenital problems, anemia, and increased burden of iron in their myocardium. This study was designed to evaluate the effects of direct acting antiviral (DAA) therapy on the cardiac function of hepatitis C virus (HCV)-infected patients with thalassemia. METHOD: HCV-infected thalassemia patients were enrolled to this prospective evaluation. Daily tablets of 90 mg Ledipasvir (or 60 mg Daclatasvir) plus 400 mg Sofosbuvir (±ribavirin) were prescribed for the patients according to the Iran Hepatitis Network's guidelines. An echocardiography fellow collected the echocardiography findings before and after the treatment of all the patients. The patients were followed up for any cardiac events within 12 weeks after finishing the treatment. RESULTS: Thirty-two patients with the mean age of 24.2 ± 6.4 years were evaluated. All patients showed a sustained virological response at the 12th week after finishing the treatment. The patients' left ventricular end systolic diameter (3.0 vs 3.24; P = 0.003) and volume (33.8 vs 43.6; P = 0.001), global longitudinal strain of the left ventricle (-22.0 vs -20.6, P = 0.046), and average (-21.4 vs -20.3; P = 0.048), and the right ventricle size (3.12 vs 3.31; P = 0.012) were significantly increased after finishing the treatment. Changes in the abovementioned parameters were not correlated with the patients' myocardium iron load. There were no significant differences in other echocardiographic parameters ( P > 0.05) before and after the treatment. CONCLUSION: Sofosbuvir-based regimens for HCV treatment were safe for our HCV-infected patients with thalassemia. Our patients' ejection fraction remained unchanged. Hence, more specialized echocardiographic evaluations were recommended for those with a history of cardiac abnormalities, cardiac iron overload, and in case of any cardiac adverse event during DAA therapy in patients with thalassemia.

First Report of Prevalence of Blood-Borne Viruses (HBV, HCV, HIV, HTLV-1 and Parvovirus B19) Among Hemophilia Patients in Afghanistan.

Blood-borne viruses including Hepatitis B and C, HIV, HTLV-1 and parvovirus B19 are still a factor of concern, especially for hemophilia patients. Although the safety of the blood supply continues to improve worldwide, the blood supply system in Afghanistan was damaged by many years of conflict and political instability. To date, there are few studies focused on the prevalence of blood-borne viruses in hemophilia patients. This study is first to investigate the prevalence of five blood-borne viruses in Afghanistan hemophilia patients in four cities including Kabul, Herat, Mazar-i-Sharif and Jalal Abad. A total of 80 hemophilia male patients were screening for the presence of five transfusion-transmitted viruses using ELISA and PCR. Data obtained showed 2.5% seropositivity for HBV, 8.75% seropositivity for HCV, and 91.25% seropositivity for parvovirus B19. None of the patients were positive for HIV and HTLV-1 and the prevalence of HCV was higher in older patients rather than younger patients. This finding, the first to report in Afghanistan, shows a high prevalence of parvovirus B19 in Afghanistan hemophilia patients and implementation of highly sensitive screening is necessary.

Morphology of the diet in the gut of threatened yellow catfish Horabagrus brachysoma (Gunther 1864) at two life stages.

The diet composition of fingerlings and adults of Horabagrus brachysoma, a threatened, freshwater catfish, endemic to South India, collected from the Periyar River of Kerala in south India was studied using standard methods. The results of the stomach content analysis revealed that the species is an omnivore, predominantly feeding on animal matter in the young stage and plant matter in the adult stage. Based on the findings of the present study, it can be concluded that H. brachysoma collected from Periyar River is a benthophagic omnivore and that adults prefer dietary fractions of plant origin over animal tissues. An ontogenic diet shift from carnivorous to more pronounced omnivorous nature is exhibited by the fish. The fish is adapted for a wide spectrum of feeding habits including the ability to digest all major types of nutrients and these findings will help to formulate a more economically viable and nutritionally balanced diet for the artificial rearing of this cultivable catfish.

Endogenous and Exogenous Natural Adjuvants for Vaccine Development.

Objective & Background: Various adjuvants are usually co-injected with an antigen for stimulation of effective immune responses. Adjuvants are able to elicit innate immune responses at the injection site. Depending on the activated type of innate responses, adjuvants can modify the quality and quantity of adaptive immune responses. Their mechanisms of action in vaccine development include: a) enhancement of the total antibody titers; b) reduction of the antigen dose; c) induction of potent cell-mediated immunity; d) increase in the speed and duration of the protective response; e) stimulation of mucosal immunity; and f) cross-protection. Up to now, different exogenous adjuvants have been identified to boost immune responses including inorganic compounds, mineral oil, bacterial products, non-bacterial organics, detergents or Quil A, plant saponins, Freund's complete or incomplete adjuvants, and delivery systems. However, some immune responses can be generated in the absence of the exogenous adjuvants. Indeed, endogenous adjuvants released from the cells were known as the danger signals and immunogenic compounds. Several main endogenous adjuvants contain cytokines, chemokines, alarmins, dendritic cells (DCs), toll like receptor (TLR) ligands or agonists, and antibodies. RESULTS & CONCLUSION: In this review, the immune activities of the natural adjuvants especially endogenous adjuvants and their mechanisms of action are discussed.

Generation of the Fluorescent HMGB1-GFP Fusion Protein in Insect Cells and Evaluation of its Immunogenicity in Two Mice Models.

BACKGROUND: High mobility group box 1 (HMGB1) is a highly conserved protein present in the nuclei and cytoplasm of cells which has an important role as a mediator of inflammation in the extracellular environment. HMGB1 was identified as an innate adjuvant that induces immune responses against soluble antigens in vivo. OBJECTIVE: Our goal is the generation of recombinant HMGB1-GFP fusion protein in insect cells for evaluation of immune responses in mouse model. METHOD: In the current study, we used a baculovirus expression system for insect cells that was based on expression of HMGB1 with target gene (GFP), and purified the recombinant HMGB1- GFP fusion protein. We then demonstrated whether immunogenicity of GFP changes in the presence or absence of recombinant HMGB1 acting as an adjuvant in C57BL/6 and BALB/c mice. RESULTS: Our data showed that HMGB1 had a major influence on antibody immune responses induced by GFP in both animal models. The groups receiving HMGB1-GFP fusion protein showed total IgG and IgG2a responses significantly higher than IgG1 in BALB/c mice. Indeed, a mixed IgG1/IgG2a response was observed with high intensity toward IgG2a. In contrast, C57BL/6 mice immunized by HMGB1-GFP protein elicited the same levels of IgG1 and IgG2a. However, the levels of IgG2a and total IgG against the recombinant GFP (rGFP) in C57BL/6 mice were lower than those in BALB/c mice. CONCLUSION: We concluded that fusion of HMGB1 with GFP was immunologically more effective than GFP alone.

Morphological and genetic evidence for multiple evolutionary distinct lineages in the endangered and commercially exploited red lined torpedo barbs endemic to the Western Ghats of India.

Red lined torpedo barbs (RLTBS) (Cyprinidae: Puntius) endemic to the Western Ghats Hotspot of India, are popular and highly priced freshwater aquarium fishes. Two decades of indiscriminate exploitation for the pet trade, restricted range, fragmented populations and continuing decline in quality of habitats has resulted in their 'Endangered' listing. Here, we tested whether the isolated RLTB populations demonstrated considerable variation qualifying to be considered as distinct conservation targets. Multivariate morphometric analysis using 24 size-adjusted characters delineated all allopatric populations. Similarly, the species-tree highlighted a phylogeny with 12 distinct RLTB lineages corresponding to each of the different riverine populations. However, coalescence-based methods using mitochondrial DNA markers identified only eight evolutionarily distinct lineages. Divergence time analysis points to recent separation of the populations, owing to the geographical isolation, more than 5 million years ago, after the lineages were split into two ancestral stocks in the Paleocene, on north and south of a major geographical gap in the Western Ghats. Our results revealing the existence of eight evolutionarily distinct RLTB lineages calls for the re-determination of conservation targets for these cryptic and endangered taxa.

Unraveling a 146 years old taxonomic puzzle: validation of Malabar snakehead, species-status and its relevance for channid systematics and evolution.

BACKGROUND: The Malabar snakehead Channa diplogramma is one of the most enigmatic and least understood species within the family Channidae, which comprise one of the most important groups of freshwater food fish in tropical Asia. Since its description from peninsular India in 1865, it has remained a taxonomic puzzle with many researchers questioning its validity, based on its striking similarity with the South East Asian C. micropeltes. In this study, we assessed the identity of the Malabar snakehead, C. diplogramma, using morphological and molecular genetic analyses, and also evaluated its phylogenetic relationships and evolutionary biogeography. METHODOLOGY/PRINCIPAL FINDINGS: The morphometric and meristic analysis provided conclusive evidence to separate C. diplogramma and C. micropeltes as two distinct species. Number of caudal fin rays, lateral line scales, scales below lateral line; total vertebrae, pre-anal length and body depth were the most prominent characters that can be used to differentiate both the species. Channa diplogramma also shows several ontogenic color phases during its life history, which is shared with C. micropeltes. Finally, the genetic distance between both species for the partial mitochondrial 16S rRNA and COI sequences is also well above the intra-specific genetic distances of any other channid species compared in this study. CONCLUSIONS/SIGNIFICANCE: The current distribution of C. diplogramma and C. micropeltes is best explained by vicariance. The significant variation in the key taxonomic characters and the results of the molecular marker analysis points towards an allopatric speciation event or vicariant divergence from a common ancestor, which molecular data suggests to have occurred as early as 21.76 million years ago. The resurrection of C. diplogramma from the synonymy of C. micropeltes has hence been confirmed 146 years after its initial description and 134 years after it was synonymised, establishing it is an endemic species of peninsular India and prioritizing its conservation value.