Several diagnostic tools are available for veterinarians and fish health professionals to evaluate fish health and their abnormalities. However, reference data regarding the character and size of fish blood cells are limited. The purpose of this study was to analyze the morphology and morphometry of normal blood cells in zebrafish (Danio rerio), common carp (Cyprinuscarpio carpio), and tilapia (Oreochromis niloticus). To get a representative sample, we took blood from ten 6-mo old healthyfish from each species. Fish were purchased from an ornamental fish market and the local government fish breeding centerin Yogyakarta, Indonesia. A total of 100 erythrocytes and a maximum of 30 leukocytes (neutrophils, eosinophils, basophils,lymphocytes and monocytes) were randomly sampled and analyzed qualitatively and quantitatively, including morphometricanalysis of both the long axis (LA) and short axis (SA) of these cells. All data obtained were analyzed using ANOVA statisticaltests to compare the blood cells in each species with SPSS software. The findings revealed distinct differences in bothmorphology and morphometry of the blood cells among the species. Basic knowledge obtained from this research will aidin the development of biomarkers and other ancillary diagnostic tools for further hematology research, conservation, andclinical diagnosis in these 3 fish species.
Tilapia lake virus disease (TiLVD) is an emerging viral disease associated with high morbidity and mortality in cultured tilapia worldwide. In this study, we have developed and validated a TaqMan quantitative reverse transcription PCR (RT-qPCR) assay for TiLV, targeting a conserved region within segment 10 of the genome. The RT-qPCR assay was efficient (mean ± SD: 96.71 ± 3.20%), sensitive with a limit of detection of 10 RNA viral copies per reaction, and detected TiLV strains from different geographic regions including North America, South America, Africa, and Asia. The intra- and inter-assay variability ranged over 0.18-1.41% and 0.21-2.21%, respectively. The TaqMan RT-qPCR assay did not cross-react with other RNA viruses of fish, including an orthomyxovirus, a betanodavirus, a picornavirus, and a rhabdovirus. Analysis of 91 proven-positive and 185 proven-negative samples yielded a diagnostic sensitivity of 96.7% and a diagnostic specificity of 100%. The TaqMan RT-qPCR assay also detected TiLV RNA in infected Nile tilapia liver tissue extracts following an experimental challenge study, and it successfully detected TiLV RNA in SSN-1 (E-11 clone) cell cultures displaying cytopathic effects following their inoculation with TiLV-infected tissue homogenates. Thus, the validated TaqMan RT-qPCR assay should be useful for both research and diagnostic purposes. Additionally, the TiLV qPCR assay returns the clinically relevant viral load of a sample which can assist health professionals in determining the role of TiLV during disease investigations. This RT-qPCR assay could be integrated into surveillance programs aimed at mitigating the effects of TiLVD on global tilapia production.
Abstract. Widiyanti R, Nugroho HA, Megarani DV, Widiasih DA, Pakpahan S. 2023. Multiplex PCR detection of mackerel-based food adulteration with pleco and chicken in selected areas around Ciliwung River, Indonesia. Biodiversitas 24: 1538-1543. Detecting fish product adulteration is crucial to ensure food safety since pleco meat was already reported to carry several heavy metals that might harm human health. Pleco is invasive species in the Ciliwung River and is commonly used as adulteration material for fish-based products. Adulteration in mackerel-based food products may alter the nutritional value and carry heavy metal contamination from the bottom-feeder fish's meat (pleco). Therefore, using the DNA barcoding technique, a molecular approach has been used to authenticate mackerel fish products (including dumplings and otak-otak). This study aimed to develop a specific multiplex PCR method for simultaneously detecting processed products from mackerel and pleco. The sample consists of 21 processed food items initially made from mackerel. The samples were taken in the selected area around the Ciliwung River. All the samples can be amplified successfully, and amplification lengths were 108, 171, and 300 bp, respectively. Analysis from various claimed mackerel products showed that five samples were positive for pleco adulteration, and 11 products contained chicken meat addition. The phylogenetic tree was constructed from selected sequences from our samples and showed that the amplicons were clustered in three clades, mackerel (Scomberomorus), pleco (Pterygoplichthys), and chicken (Gallus gallusLinnaeus, 1758). The findings of this study revealed that 23.80% (5/21) products contained pleco, and 52.38% (11/21) contained chicken meat addition. The addition of an unusual component to food composition may alter nutritional value as well as may affect food hygiene and safety.
Ultraviolet (UV) lamp is the simplest method for sterilizing operating theatre. This method is effective, easily operated, and does not require high cost. Furthermore, there were several studies of microorganism contamination in the air and surface at human operating theatre. However, studies in veterinary operating theatre related to the effectiveness of UV light on sterilization process is still limited, especially in Indonesia. Bacterial contamination samples were collected three times each in three different conditions: A) before surgery and without UV, B) before surgery but UV was already used, and C) after surgery and UV was already used. Samples were taken with settle plate and swab method for collecting the air and operating table contamination, respectively. One-way repeated measures ANOVA determined that there was statistically significant difference in the number of bacterial contaminations between three conditions (A, B, and C) in settle plate method (p=0.009), as well as in swab method (p=0.010). The result revealed that the UV light was effective to sterilize operating theatre, which can be seen from the significant decreases on the number of bacterial contaminations before and after the UV was used, both in settle plate and swab method. The result of this study supported the theory that the UV light can reduce the air bacterial and surface contamination at operating theatre. However, the result of microorganism contaminations in this study was still not appropriate based on the standard minimum of total bacterial in the operating theatre from The Ministry of Health, Republic of Indonesia. Consequently, the use of another method of sterilization at the operating theatre is still required for a better sterilization result.
Abstract. Widayanti R, Nugroho HA, Megarani DV, Widiasih DA, Pakpahan S. 2021. Revealing Spanish mackerel’s diversity in Indonesian through local commodities in the fish market. Biodiversitas 23: 624-630. The objective of this study was to explore the diversity of Spanish mackerel in Indonesian Archipelago based on commodities offered in local fish markets. Eighteen specimens were collected from six different fish markets around the Indonesian archipelago. According to Wallace Line, the cytochrome B sequence was used as a genetic marker to reveal diversity from West Indonesia to East Indonesia. Gene amplification was performed using the polymerase chain reaction followed by Sanger sequencing. Based on DNA sequence analysis, we identified three species of Spanish Mackerel available at various fish markets around the region. The first group is related to Scomberomorus commerson, which was sold in both Western and Eastern Indonesian fish markets; the second group is related to Scomberomorus semifasciatus, which was sold in Eastern Indonesian fish markets; and the third group is related to Scomberomorus koreanus, which was sold in Western Indonesian fish markets. The genetic differences amongst Spanish Mackerel populations ranged from 0 to 17% and the average evolutionary divergence in the overall populations was determined to be 4%. The data collected can be utilized to initiate map mackerel diversity based on CYT B sequence throughout the Indonesian Archipelago, as well as for further study and application in the identification of foods produced from mackerel.
Most of raptor species rehabilitated for conservation purpose at WRC Jogja are eagles. Annually, we perform general raptor examination involving conventional routine hematologic examination. However, low sample quality and over dense blood cell population during total erythrocyte and leukocyte counting were deemed as obstacles. This study aimed to give wider insight in how total erythrocyte and leukocyte counting was performed at WRC Jogja to overcome those obstacles. Firstly, handling and restraint were done physically adapting the Aba method. One-to-two half millilitres of blood was collected from the ulnar vein using 3-ml-syringe with 24-gauge-needle which was then preserved in an EDTA-containing tube and proceeded to subsequent method no more than 3 hours after collection. No hypovolemic shock was observed. During total erythrocyte and leukocyte counting using Neubauer-ruled counting chamber, we modified the dilution factor to be 250-fold and 50-fold respectively. This method yielded a countable cell density and can be applied in other species in similar condition.
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