Ontogeny of digestive enzymes in larvae of the Clown Anemonefish, Amphiprion ocellaris (Perciformes: Pomacentridae)

Introduction: The Clown anemonefish (Amphiprion ocellaris) is the most popular fish species in the marine aquarium trade; however, there is a lack of information on their digestive physiology during larval ontogeny, valuable information needed for diet design and management protocols. Objective: To characterize the early digestive enzymes of A. ocellaris larvae. Methods: We used three pools (10 larvae each) and extracted 10 samples per tank, from just before hatching to the 38th day. We analyzed the specific activity of acid and alkaline proteases, trypsin, chymotrypsin, leucine aminopeptidase and lipase; and did acid and alkaline protease zymograms. Results: We detected all measured enzymes at hatching. Acid proteases increased in activity until the 38th day. Alkaline proteases, trypsin, chymotrypsin, and leucine aminopeptidase had the same pattern, and maximum activity on the 8th day, decreasing at the 38th day. Lipase activity peaked on the 8th and 30th day. The acid zymogram had a single band, appearing on the 8th day. A total of eight alkaline proteases were revealed (154.2, 128.1, 104.0, 59.8, 53.5, 41.9, 36.5 and 25.1 KDa), with seven bands on the 1st day and all bands from the 3rd to 8th day, decreasing at two bands (41.9 and 25.1 KDa) in the 38th day. Conclusion: A. ocellaris has a functional stomach on the 8th day, and, on the 38th day, a digestive omnivore pattern with a tendency to carnivory.

Introducción: El pez payaso (Amphiprion ocellaris) es la especie de pez más popular en el comercio de acuarios marinos; sin embargo, falta información sobre su fisiología digestiva durante la ontogenia larval, información valiosa necesaria para protocolos de diseño y manejo dietético. Objetivo: Caracterizar las enzimas digestivas tempranas de larvas de A. ocellaris. Métodos: Usamos tres homogenados (con 10 larvas cada uno) y extrajimos 10 muestras por tanque, justo antes de la eclosión hasta el día 38. Analizamos la actividad específica de proteasas ácidas y alcalinas, tripsina, quimotripsina, leucina aminopeptidasa y lipasa; e hicimos zimogramas de proteasas ácidas y alcalinas. Resultados: Detectamos todas las enzimas medidas en la eclosión. La actividad de proteasas ácidas incrementó hasta el día 38. Proteasas alcalinas, tripsina, quimotripsina, y leucina aminopeptidasa tuvieron el mismo patrón, con actividad máxima en el octavo día, decreciendo en el día 38. Hubo picos en la actividad lipasa a los ocho y 30 días. El zimograma ácido tuvo una banda única, apareciendo al octavo día. Se hallaron ocho proteasas alcalinas (154.2, 128.1, 104.0, 59.8, 53.5, 41.9, 36.5 y 25.1 KDa), con siete bandas al primer día, y todas las bandas entre el tercer y octavo día, bajando a dos bandas (41.9 y 25.1 KDa) al día 38. Conclusión: A. ocellaris tiene un estómago funcional al octavo día, y, al día 38, un patrón digestivo omnívoro con tendencias carnívoras.

High frequency of larval abnormalities of the toad Rhinella arenarum (Anura: Bufonidae) in an Argentinian agroecosystem

Introduction: The frequent use of pesticides is currently considered a cause of environmental pollution due to the high rate of entry of these substances into agroecosystems. This constitutes a risk for the species that inhabit these ecosystems, in particular anurans whose characteristics make them prone to exposure to and interaction with environmental pollutants. Objective: To report the occurrence of abnormalities in larvae of the common toad Rhinella arenarum inhabiting ponds surrounded by agroecosystems. Methods: In two consecutive springs (2015 and 2016), reproductive events of common toads were monitored in temporary pond systems in agricultural and non-agricultural areas, located near the city of La Plata (Buenos Aires, Argentina). The physicochemical parameters of the ponds were measured, and the stage of each reproductive event was recorded, such as the numbers of adult toads, amplexus and clutches. In the laboratory, the larvae were measured and photographed, their stage of development was recorded, and their morphology was examined under a stereomicroscope. Representative samples (normal and abnormal) from each pond studied were processed for histopathological analysis. Results: In the field studies carried out on a population of R. arenarum collected in an agroecosystem, a lower number of reproductive adults and clutches were observed in relation to the population of a non-agricultural pond. A total of 1 910 larvae were collected: 529 and 1 381 larvae from ponds located in non-agricultural and agricultural areas, respectively. Larvae from the agroecosystem showed two types of abnormalities: severe tail flexure and abdominal bloating. In addition, five degrees of severity could be determined in relation to abdominal bloating. Conclusions: This work reports the high frequency and severity of abnormalities observed in the early stages of R. arenarum larvae living within an agroecosystem, providing evidence of the negative impact that agricultural activities cause on aquatic ecosystems surrounded by farming areas.

Introducción: El uso frecuente de plaguicidas es considerado actualmente una causa de contaminación ambiental debido a las altas tasas de ingreso de estas sustancias a los agroecosistemas. Esta situación es un riesgo para las especies que habitan en estos ecosistemas, en particular los anuros cuyas características los hacen propensos a la exposición e interacción con contaminantes ambientales. Objetivo: Informar la presencia de anormalidades en larvas del sapo común Rhinella arenarum que habitan en estanques rodeados por un agroecosistema. Métodos: En dos primaveras consecutivas (2015 y 2016), se monitorearon los eventos reproductivos del sapo común proveniente de sistemas de estanques temporales ubicados en zonas agrícolas y no agrícolas, cerca de la ciudad de La Plata (Buenos Aires, Argentina). Se midieron los parámetros fisicoquímicos de los estanques y se registraron las etapas de cada evento reproductivo como el número de sapos adultos, amplexos y nidadas. En el laboratorio, las larvas fueron medidas y fotografiadas, se registró su estado de desarrollo y se examinó la morfología de cada una bajo microscopio estereoscópico. Se procesaron muestras representativas (normales y anormales) de cada estanque estudiado para análisis histopatológico. Resultados: En la población de R. arenarum que vive dentro de un agroecosistema, se observó un menor número de adultos reproductores y puestas en relación con la del estanque en la zona no agrícola. Se recolectaron un total de 1 910 larvas: 529 y 1 381 larvas de estanques ubicados en zonas no agrícolas y agrícolas, respectivamente. Las larvas del agroecosistema mostraron dos tipos de anormalidades: severa flexión de la cola y distensión abdominal. Además, se pudo determinar cinco grados de gravedad en relación con la distensión abdominal. Conclusiones: Una alta frecuencia y severidad de anormalidades en los estadios tempranos de larvas de R. arenarum que viven dentro de un agroecosistema proporciona evidencia del impacto negativo que las actividades agrícolas causan en los ecosistemas acuáticos rodeados por áreas de cultivo.

Effect of glyphosate and ciprofloxacin exposure on enteric bacteria of tadpoles / Efecto de la exposición a glifosato y ciprofloxacina en bacterias entéricasde renacuajos

Abstract The high load of agrochemicals and antibiotics present in agricultural aquatic environments represents a risk for wildlife. Since enteric bacteria, which play a key role in the physiological functioning of their hosts, are sensitive to a wide variety of pollutants, their study allows to evaluate the health of organisms. This study aimed to evaluate the effects of commercial formulations of a glyphosate-based herbicide (GBH) and the antibiotic ciprofloxacin (CIP), individually and in mixture, on the bacterial diversity of the intestinal content of common toad (Rhinella arenarum) tadpoles. The diversity of cultivable fast-growing bacteria with low nutritional requirements was evaluated using classic microbiological tests and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry identification. Bacterial diversity varied among treatments. Taxa diversity increased in the GBH-treated group but decreased in the CIP-treated group. Remarkably, Yersinia spp. and Proteus spp. were only found in the GBH-treated group. The prevalence of Klebsiella spp. and Pseudomonas spp. decreased in the intestinal microbiota of the GBH-CIP-treated group. To our knowledge, this is the first report on the alteration of cultivable enteric bacteria of autochthonous tadpoles due to two pollutants of emerging concern. Our results demonstrate that R. arenarum tadpoles can be used as non-conventional model organisms for environmental pollution monitoring. Our preliminary findings would contribute to understanding how the presence of GBH and CIP in freshwaters may represent a threat to wildlife and human health by causing enteric dysbiosis of part of the bacterial community.

Resumen La alta carga de agroquímicos y antibióticos en los ambientes acuáticos y los agroe-cosistemas representa un riesgo para la vida silvestre. Dado que la microbiota intestinal juega un papel fundamental en el funcionamiento de su hospedador y es sensible a una amplia variedad de contaminantes, su estudio permite evaluar la salud de los organismos. En este trabajo estudiamos los efectos de formulaciones comerciales de un herbicida a base de glifosato (GBH) y del antibiótico ciprofloxacina (CIP), por separado y en mezcla, sobre la diversidad de bacterias intestinales de renacuajos del sapo común (Rhinella arenarum). El estudio de la diversidad de bacterias entéricas cultivables de rápido crecimiento y bajo requerimiento nutricional se llevó a cabo utilizando pruebas microbiológicas clásicas e identificación por espectrometría de masas de tiempo de vuelo por desorción/ionización láser asistida por matriz (MALDI-TOF). La microbiota entérica fue diferente según el tratamiento. El GBH indujo un aumento de la diversidad bacteriana, mientras que la CIP produjo una reducción. Entre estos cambios, destaca la presencia de Yersinia spp. y Proteus spp. solo en el tratamiento con GBH. Además, en el tratamiento GBH-CIP se encontró una disminución en la prevalencia de Klebsiella spp. y Pseudomonas spp. en la microbiota intestinal de los renacuajos. Este es el primer informe sobre la alteración del contenido bacteriano intestinal de renacuajos de R. arenarum producido por dos contaminantes emergentes de preocupación. Demostramos que el renacuajo del sapo común se puede utilizar como un organismo modelo no convencional para el monitoreo de la contaminación ambiental. Estos hallazgos constituyen el primer paso para comprender cómo la presencia de GBH y CIP en aguas dulces puede representar una amenaza para la vida silvestre y la salud humana a través de la disbiosis entérica asociada al efecto sobre la comunidad bacteriana.

The chronicle of the ups and downs that made Bacillus thuringiensis a natural insecticide

Bacillus thuringiensis is a worldwide known bacterium for its capacity to control insect pests thanks to the action of its parasporal crystal. The objective of this paper deals with the history, in some cases unknown, of the study of Bacillus thuringiensis that led it to be a crucial biological alternative in controlling pest insects. How the mode of action for killing insects was understood, as well as the field tests that were carried out to evaluate its effectiveness and to develop the first commercial products, are reflected in this review that presents and discusses the scientific successes and failures that marked the course of B. thuringiensis.

Bacillus thuringiensis es una bacteria conocida mundialmente por su capacidad para controlar insectos plaga, gracias a la acción de su cristal parasporal. El objetivo de esta revisión trata de la historia, en algunos casos desconocida, del estudio de Bacillus thuringiensis que la llevó a ser una importante alternativa biológica en el control de insectos plaga. Cómo se llegó a comprender el modo de acción para matar insectos, así como las pruebas de campo que se realizaron para evaluar su efectividad y lograr desarrollar los primeros productos comerciales están plasmados en esta revisión que presenta y discute los aciertos y desaciertos científicos que marcaron el rumbo de B. thuringiensis.

Settlement and post-settlement survival of Orbicella annularis and Orbicella faveolata (Scleractinia: Merulinidae) on substrates with coatings

Introduction: One of the main bottlenecks in restoration projects based on sexual reproduction is post-settlement survival, mainly due to competition for substrate with fleshy algae and predation. Therefore, substrates of different shapes and materials have been created and tested, seeking to optimize these processes with attractive surfaces for the larvae and structures where the recruits are protected from predation, and competition is reduced. Objective: To improve settlement and post-settlement survival of two important Caribbean reef-building corals, using different coatings on substrates. Methods: To determine whether substrate coatings properties are favourable to larval settlement in Orbicella annularis, and O. faveolata, collected in Puerto Morelos, Mexican Caribbean, we evaluated their settlement for three weeks on six coatings with a combination of properties. Each coating was designed to provide a combination of two out of three properties: 1) water repellence (hydrophobicity), 2) phosphorescence-based colour, and 3) mineral-enriched surface chemistry. In a separate experiment larvae settlement was tested using coatings with a single property. Finally, we determined the post-settlement survival of O. annularis and O. faveolata on the different coatings for seven weeks. Results: The combination of high hydrophobicity and light blue phosphorescent microparticles and high hydrophobicity and red-orange phosphorescent microparticles resulted in a higher settlement of O. annularis and O. faveolata when compared with other coatings (30.8 - 66.7 % higher). No significant differences were found in the number of larval settled when the water-repellence and the phosphorescence-based were evaluated independently. Post-settlement survival time on substrates was low, with a maximum of 34 days after settlement for O. annularis and 42 days for O. faveolata. Conclusions: In terms of the larval settlement, the combination of the coatings properties appears to play an essential role in the choice of microhabitat for both O. annularis and O. faveolata. But individually these properties did not generate an advantage in the larval settlement. Moreover, some chemical components associated with the coatings may be counterproductive to the survival of the polyps over time.

Introducción: Uno de los principales cuellos de botella en proyectos de restauración basada en reproducción sexual es la supervivencia de las larvas posterior al asentamiento, principalmente por la competencia por el sustrato con algas filamentosas y la depredación. Por ello, se han creado y analizado sustratos de diferentes formas y materiales, buscando optimizar estos procesos con superficies atrayentes para las larvas, y estructuras donde los reclutas se encuentran protegidos de la depredación y se disminuya la competencia. Objetivo: Mejorar el asentamiento y la supervivencia de dos importantes corales formadores de arrecifes del Caribe, utilizando diferentes recubrimientos en sustratos. Métodos: Para determinar si las propiedades de la superficie del sustrato son favorables para el asentamiento de larvas de Orbicella annularis y O. faveolata, recolectadas en Puerto Morelos, Caribe mexicano, evaluamos su asentamiento durante tres semanas en seis recubrimientos con una combinación de propiedades. Cada recubrimiento fue diseñado para proporcionar una combinación de dos de tres propiedades: 1) repelencia al agua (hidrofobicidad), 2) fosforescencia y 3) química superficial enriquecida con minerales. En un experimento separado se evaluó el asentamiento de larvas en sustratos con recubrimientos de una sola propiedad. Finalmente, se determinó la supervivencia posterior al asentamiento de O. annularis y O. faveolata sobre los diferentes recubrimientos durante siete semanas. Resultados: La combinación de alta hidrofobicidad y micropartículas fosforescentes azules y alta hidrofobicidad y micropartículas fosforescentes rojo-naranja dio como resultado un mayor asentamiento de O. annularis y O. faveolata en comparación con otros recubrimientos (30.8 - 66.7 % mayor). No se encontraron diferencias significativas en el número de larvas asentadas cuando se evaluaron de forma independiente la repelencia al agua y la fosforescencia. El tiempo de supervivencia posterior al asentamiento en los sustratos fue bajo, con un máximo de 34 días después del asentamiento para O. annularis y 42 días para O. faveolata. Conclusiones: En el asentamiento de larvas, la combinación de las propiedades del recubrimiento parece desempeñar un papel importante en la elección del microhábitat tanto para O. annularis como para O. faveolata. Pero de forma individual estas propiedades no generaron una ventaja en el asentamiento larvario. Además, algunos componentes químicos asociados con los recubrimientos pueden ser contraproducentes para la supervivencia de los pólipos a lo largo del tiempo.

Interpretation of Detection and identification standard of hookworm—Hook-worm larvae coproculture techniques (WS/T 791—2021) / 中国血吸虫病防治杂志

Detection and identification standard of hookworm—Hookworm larvae coproculture techniques (WS/T 791—2021) is the first recommended technical standard for hookworm detection and species identification using the hookworm larvae coproculture technique in China. This standard was issued on November 23, 2021, and had been in effect since May 1, 2022. This article provides a detailed interpretation pertaining to the background, drafting process, main contents, and dos and don’ts for better understanding and application of this standard among professionals working in disease control and prevention institutions and medical institutions.

Density Of Eggs and Larvae of Aedes Spp and the Characteristics of their Larvae Habitat in Endemic Dengue Area of Ternate City

@#Introduction: Dengue Fever (DF) is a disease spread by Aedes spp. caused by dengue virus infection. The study aimed to identify the density of Aedes spp eggs and larvas stages and the characteristics of their habitat in dengue-endemic areas in Ternate City in September-December 2021. Methods: the research was conducted in four dengue-endemic villages in Ternate City, namely Sangaji, Maliaro, Bastiong Karance, and Bastiong Talangame village. In 80 households, egg density was determined by placing 2 ovitraps in each house, one inside and one outside the house. After a week, the filter paper which is a medium mosquito to lay their eggs in the ovitrap was collected, and the ovitrap index value was calculated. The value of the House Index (HI), Container Index (CI), and Breuteu Index (BI) was used to calculate the larval density. The type and materials of the containers were used to assess the parameters of the larval environment. Results: The ovitrap index value in the four villages was categorized as moderate level 3 (27.50% -36.25%). The highest larval density was found in Sangaji Village (HI = 81%), while the lowest was in Bastion Talangame Village (HI = 70%). The highest CI and BI values were found in Bastiong Karance Village (CI=51.5% and BI=190%), and the lowest was in Maliaro Village (CI=37.5% and BI=128%). Density figures in all endemic villages have a high larval density with a value of 8. Conclusion: There was no significant difference (p>0.05) between the types and the materials of containers in each village in Dengue endemic areas. The high density of the egg and larval stage and the information characteristic of habitat Aedes spp. could be considered as basic information for dengue vector control in Ternate City.

Acute Developmental Toxicity of Panax notoginseng in Zebrafish Larvae / 中国结合医学杂志

OBJECTIVE@#To evaluate toxicity of raw extract of Panax notoginseng (rPN) and decocted extract of PN (dPN) by a toxicological assay using zebrafish larvae, and explore the mechanism by RNA sequencing assay.@*METHODS@#Zebrafish larvae was used to evaluate acute toxicity of PN in two forms: rPN and dPN. Three doses (0.5, 1.5, and 5.0 µ g/mL) of dPN were used to treat zebrafishes for evaluating the developmental toxicity. Behavior abnormalities, body weight, body length and number of vertebral roots were used as specific phenotypic endpoints. RNA sequencing (RNA-seq) assay was applied to clarify the mechanism of acute toxicity, followed by real time PCR (qPCR) for verification. High performance liquid chromatography analysis was performed to determine the chemoprofile of this herb.@*RESULTS@#The acute toxicity result showed that rPN exerted higher acute toxicity than dPN in inducing death of larval zebrafishes (P<0.01). After daily oral intake for 21 days, dPN at doses of 0.5, 1.5 and 5.0 µ g/mL decreased the body weight, body length, and vertebral number of larval zebrafishes, indicating developmental toxicity of dPN. No other adverse outcome was observed during the experimental period. RNA-seq data revealed 38 genes differentially expressed in dPN-treated zebrafishes, of which carboxypeptidase A1 (cpa1) and opioid growth factor receptor-like 2 (ogfrl2) were identified as functional genes in regulating body development of zebrafishes. qPCR data showed that dPN significantly down-regulated the mRNA expressions of cpa1 and ogfrl2 (both P<0.01), verifying cpa1 and ogfrl2 as target genes for dPN.@*CONCLUSION@#This report uncovers the developmental toxicity of dPN, suggesting potential risk of its clinical application in children.

Internal ultrastructure of the planktonic larva of the coral Porites panamensis (Anthozoa: Scleractinia) / Ultraestructura interna de la larva planctónica del coral Porites panamensis (Anthozoa: Scleractinia)

Abstract Introduction: The scleractinian coral life cycle includes planktonic larvae that settle on the benthos, allowing the primary polyp to clone and build a sexually reproducing adult colony. The larval physiology and ecology of Eastern Tropical Pacific scleractinians needs the exploration of basic aspects such as the internal morphology of planulae. Objective: To describe histological and cytological characteristics of Porites panamensis larvae. Methods: During August-July 2019, at Islas Marias Biosphere Reserve, Central Mexican Pacific, we made 14 collections of coral larvae and identified the species with cytochrome oxidase subunit 1 gene. We used a scanning electron microscope and other techniques. Results: The ectoderm was composed by heterogeneous, mono-ciliated, columnar epithelial cells. Nematocysts were clustered at the oral pole of the ectoderm, and cells were evident in the aboral pole of the ectoderm gland. The endoderm had secretory cells, lipids and symbionts. Conclusions: The abundance of secretory cells and nematocysts in the aboral pole suggests their importance in substrate exploration and larval settlement. Our results support previous descriptions of larval ultrastructure in other coral species.

Resumen Introducción: El ciclo de vida del coral escleractinio incluye larvas planctónicas que se asientan en el bentos, lo que permite que el pólipo primario se clone y construya una colonia de adultos con reproducción sexual. La fisiología y ecología larvaria de los escleractinios del Pacífico Tropical Oriental necesita la exploración de aspectos básicos como la morfología interna de las plánulas. Objetivo: Describir las características histológicas y citológicas de las larvas de Porites panamensis. Métodos: Durante agosto-julio 2019, en la Reserva de la Biosfera Islas Marías, Pacífico Central Mexicano, realizamos 14 recolectas de larvas de coral e identificamos las especies con el gen citocromo oxidasa subunidad 1. Utilizamos un microscopio electrónico de barrido y otras técnicas. Resultados: El ectodermo está compuesto por células epiteliales columnares heterogéneas, monociliadas. Los nematocistos se agrupan en el polo oral del ectodermo, mientras que en el polo aboral son visibles células glandulares. El endodermo presentó células secretoras, lípidos y simbiontes. Conclusiones: La abundancia de células secretoras y nematocistos en el polo aboral sugiere su importancia en la exploración del sustrato y asentamiento larvario. Nuestros resultados respaldan las descripciones previas de la ultraestructura de las larvas en otras especies de coral.

Effect of Eosin Solution Exposed to Sunlight against Anopheles Mosquito Larvae

Mosquitoes are a menace for millions of people around the world, they are vectors for destructive microorganisms which cause diseases like Malaria, Dengue fever and Lymphatic filariasis, affecting people in developing countries and areas with tropical climates. Anopheles, a predominant genus transmits malaria, and the World Health Organization have shown that 350,000 lives especially children and pregnant women are lost annually by it. The aim of this study is to determine the effect of eosin solution exposed to sunlight on larvae of Anopheles mosquito. The use of control method was aid at eliminating the larva stages of the mosquito life cycle. A total of one hundred and twenty (120) Anopheles mosquito larvae were harvested using dipper with handle and net from drainages at Eagle Island and Rivers State University both in Port Harcourt. Five different concentrations of Eosin solutions were prepared in volumes of 1000 microlitre (µl), 800µl, 600µl, 400µl, 200µl after a stock solution of 1gram(g) in 100ml and a control, the physicochemical parameters of the solutions were determined using Extech model DO700 measuring instrument. Twenty (20) mosquito larvae, were carefully introduced into each of the concentrations, exposed to sunlight and observed for 24 hours (hrs) for a period of six (6) days for susceptibility. A hundred percent (100%) mortality was recorded in eosin volume of 1000µl and 800µl. The separate solutions of eosin showed significant effects of their concentrations on the Anopheles mosquito larvae of P-value 0.017 at P<0.05. The result obtained for the physicochemical parameters were; pH 5.24, temperature 30.4oC, conductivity 168µS/cm, salinity 0.08%, total dissolved solids 118 milligram per litre(mg/L) and dissolved oxygen was 6.5mg/L for the control. Changes occurred in the values of the dissolved oxygen before and after exposure to sunlight in all the dilutions. The results obtained showed that after 24 hrs, the mortality rate of the larvae increased, indicating that Anopheles mosquito larvae expose to concentrations of eosin solutions results in their mortality within 48 hrs. It may be concluded that this study has provided some evidence of larvicidal effect of eosin solution exposed to sunlight on larvae of Anopheles mosquito.

Anopheles gambiae Larval Development and Toxicity抯 Reduction of Conventional Agriculture Insecticides in the Laboratory Conditions

Background: Vector control of Malaria is mainly made by using impregnated bed nets and insecticides pulverizations indoor or/and outdoor. Besides, appearance and development of resistance抯 phenomenon among mosquito populations to insecticides, constitute a significant obstacle this fighting. Aims: To highlighting a neutralization phenomenon of three insecticides (methyl-parathion, dimethoate and cypermethrin) during development of the Anopheles gambiae s.s. larvae. Methodology: Two setups followed one after the other were designed. In setup 1, four concentrations (with four replicates each) were freshly prepared and independently received a first batch of 100 first instars An. gambiae s.s. After emergence of adults from this first batch, the same test media were simply filtered and received a second batch of first instars larvae to make setup 2. Three endpoints were measured in this study: the duration of larval phase, the larval mortality, and the size of adults. Results: The development duration and mortality of larvae decreased significantly at setup 2 with cypermethrin and methyl-parathion. Thus, the duration of larval stage decreased from 10.18 days at setup 1 to 7.84 days at setup 2 for 0.010 礸/l (highest concentration) with cypermethrin and from 10.20 days at setup 1 to 8.27 days at setup 2 for 0.144 礸/l (highest concentration) with methyl-parathion. The larval mortality dropped from 79.32 % at setup 1 to 12.00 % at setup 2 for the highest concentration of cypermethrin and from 76.42 % at setup 1 to 12.50 % at setup 2 for the highest concentration of methyl-parathion. While adults size significantly increased in setup 2. For males, wing抯 length increased from 3.28 mm at setup 1 to 3.49 mm at setup 2 for the highest concentration of cypermethrin, from 3.31 mm at setup 1 to 3.49 mm at setup 2 for the highest concentration of methyl-parathion. In female, wing抯 length increased from 3.52 mm at setup 1 to 3.68 mm at setup 2 for the highest concentration of cypermethrin, from 3.49 mm at setup 1 to 3.68 mm at setup 2 for the highest concentration of methyl-parathion. Conclusion: This work shows that mosquito larvae, especially An. gambiae are able to modify breeding medium to improve its fitness during their development.

Primer reporte en Cuba de Anisakis spp. en muestras de pescado en conserva / First report in Cuba of Anisakis spp. in samples of canned fish

Introducción: Las especies de Anisakis representan uno de los grupos parasitarios patógenos transmitidos por alimentos más importantes en relación con sus capacidades zoonóticas y pueden producir una enfermedad emergente llamada anisakiosis. En Cuba, no existen reportes de la enfermedad como tampoco publicaciones sobre la detección del parásito en muestras de productos pesqueros. Objetivo: Identificar larvas de Anisakis spp. en muestras de productos pesqueros. Métodos: Se realizó un estudio para la búsqueda de larvas de Anisakis spp. en muestras de productos pesqueros en conserva recibidos en el Laboratorio de Parasitología de los alimentos del Instituto Nacional de Higiene, Epidemiología y Microbiología de Cuba. Se recibieron 96 muestras de dos marcas diferentes. A cada marca se le asignó las letras A o B para proteger la identidad del fabricante. Las larvas recuperadas fueron previamente aclaradas con lactofenol e identificadas morfológicamente por microscopía. Se utilizaron las claves de identificación para larvas de Anisakis spp. descritas en la literatura. Resultados: El 38,5 por ciento del total de muestras de pescado en conserva analizadas estuvieron parasitadas con larvas de Anisakis spp., con una frecuencia media de 1,2 larvas por cada submuestra. Las larvas fueron clasificadas de tercer estadio y de tipo I. Conclusiones: Por primera vez se identificó en Cuba larvas de Anisakis spp. en muestras de pescado en conserva y se proporcionó datos de interés a las autoridades sanitarias para el fortalecimiento de políticas públicas relacionadas con la calidad e inocuidad de los alimentos(AU)

Introduction: Anisakis species represent one of the most important foodborne pathogenic parasitic groups in relation to their zoonotic capabilities, and can produce an emerging disease called anisakiosis. In Cuba, there are no reports of the disease and there are no publications on the detection of the parasite in samples of fishery products. Objective: To identify Anisakis spp. larvae in samples of fishery products. Methods: A study was conducted to search for Anisakis spp. larvae in samples of canned fish products received at the Laboratory of Food Parasitology of the National Institute of Hygiene, Epidemiology and Microbiology of Cuba. Ninety-six samples of two different brands were received. Each brand was assigned letter A or B to protect the identity of the manufacturer. The larvae recovered were previously rinsed with lactophenol and identified morphologically by microscopy. The identification keys for Anisakis spp. larvae described in the literature were used. Results: 38.5 percent of the total canned fish samples analyzed were parasitized with Anisakis spp. larvae, with a mean frequency of 1.2 larvae per subsample. The larvae were classified as third instar and type I. Conclusions: For the first time in Cuba, Anisakis spp. larvae were identified in canned fish samples and provided data of interest to health authorities to strengthen public policies related to food quality and safety(AU)

Vertical migration of Haemonchus sp. infectant larvae on Stylosanthes spp. and Brachiaria brizantha (Syn. Urochloa brizantha) cv. Marandu forage

The present work aimed to evaluate the influence of Stylosanthes spp. (Campo Grande© styles) and Brachiaria brizantha cv. Marandu in the vertical migration of Haemonchus spp. infective larvae, in the region of Uberlândia, Minas Gerais. Seeds of Stylosanthes spp. and Brachiaria brizantha cv. Marandu were planted in 50 pots of five liters (20 cm diameter) evenly disposed in an area of the Capim Branco Experimental Farm, Federal University of Uberlândia. When both species reached the minimum height of 20 cm, the pots were experimentally contaminated with sheep feces coprocultures. Samplings were performed, with complete removal of the vegetal material contained in the pots, on days one, three, six, 10 and 14 after contamination. Nematoids were recovered from the plant samples by thermo and hydrotropism, counted under an optical microscope and identified. For statistical analysis, the data were analyzed in a completely randomized design (DIC) in a 2X5 factorial scheme (two fodder species X 5 collection dates). No effect of the forage species was observed on the number of L3 of Haemonchus sp. recovered in the feces (p = 0.7790), upper stratum (p = 0.1755) or lower stratum (p = 0.7883) of the forage, however, there was a significant decrease in the number of L3 found with the passing of the days. When comparing the mean L3 recovered in the upper (0.06±0.31 L3) and lower (2.94±0.39) strata, a higher number of larvae was observed in the lower stratum (p < 0.0001). Therefore, in both forage species studied, the upper stratum of the plants, preferably pastured by the animals, remained with less contamination of infective larvae and the contamination tended to decrease over the days.

Efectividad del temefos 1 % en larvas de Culex quinquefasciatus. Matanzas, 2019

RESUMEN Introducción: el temefos es el producto más utilizado para el tratamiento focal. Existe en diferentes formulaciones y se aplica para los depósitos de agua potable, en forma de gránulos de arena a una concentración del 1 %. Objetivo: determinar la duración de la efectividad del temefos, en su formulación costarricense Biolarv G1, en una población matancera de Culex quinquefasciatus. Materiales y métodos: se realizó un bioensayo de laboratorio, utilizando la F1 de una cepa matancera de Culex quinquefasciatus, la que se sometió a los efectos del Biolarv G1, lote 1180829. Se utilizaron tres variantes de recambio de agua potable. Resultados: la efectividad fue del 100 % de mortalidad larvaria hasta el 7º día en el recambio del 100 % de agua. Para el recambio del 50 % de agua, la mortalidad promedio del 1º al 17º día fue del 97,5 %. En el caso del recambio del 30 % de agua, la mortalidad fue del 100 % hasta el 27º día. Conclusiones: se demostró que el temefos, en su formulación Biolarv G1, puede tener mayor efectividad en recambios del 30 % de agua, con una durabilidad de alrededor de 30 días, lo que propicia la protección de los depósitos durante los ciclos de trabajo diseñados para la vigilancia y lucha antivectorial en Cuba.

ABSTRACT Introduction: temefos is the most widely used product for focal treatment. It exists in different formulations and is applied for drinking water tanks, in the form of sand-like granules at a concentration of 1 %. Objective: to determine the duration of temefos effectiveness, in its Costa Rican formulation Biolarv G1, in a Matanzas population of Culex quinquefasciatus. Materials and methods: a laboratory bioassay was performed, using the F1 of a Matanzas strain of Culex quinquefasciatus, which was subjected to the effects of Biolarv G1, lot 1180829. Three variants of drinking water replacement were used. Results: the effectiveness was 100% of larval mortality until the 7th day when 100 % of wáter was replaced. For the replacement of 50 % of water, average mortality was 97.5 % from the 1st to the 17th day. In the case of 30 % water replacement, mortality was 100 % up to the 27th day. Conclusions: it was shown that temefos, in its formulation Biolarv G1, can be more effective in replacements of 30 % of water, with a durability of about 30 days, which facilitates the protection of reservoirs during work cycles designed for surveillance and anti-vector control in Cuba.

Density Of Eggs and Larvae of Aedes Spp. and the Characteristics of their Larvae Habitat in Endemic Dengue Area In Ternate City

@#Introduction: Dengue Fever (DF) is a disease spread by Aedes spp. caused by dengue virus infection. The study aimed to identify the density of egg and larval Aedes spp. stages and the characteristics of their habitat in dengue-endemic areas in Ternate City in September 2021. Methods: the research was conducted in four dengue-endemic villages in Ternate City, including Sangaji, Maliaro, Bastiong Karance, and Bastiong Talangame. In 80 households, egg density was determined utilizing ovitrap placement of up to two pieces per dwelling, one inside and one outside the house. After a week of ovitrap installation, the filter paper was collected, and the ovitrap index value was calculated. The value of the House Index (HI), Container Index (CI), and Breuteu Index (BI) was used to calculate the larval density. The type and materials of the containers were used to assess the parameters of the larval environment. Results: The ovitrap index value in the four sub-districts was categorized as moderate level 3 (27.50% -36.25%). The highest larval density was found in Sangaji Village (HI = 81%), while the lowest was in Bastion Talangame Village (HI = 70%). The highest CI and BI values were found in Bastiong Karance Village (CI=51.5% and BI=190%), and the lowest was in Maliaro Village (CI=37.5% and BI=128%). Density figures in all endemic villages have a high larval density with a value of 8. Conclusion: There was no significant difference (p>0.05) between the types and the materials of containers in each village in DF endemic areas. The high density of the egg and larval stage and the information characteristic of habitat Aedes spp. Could be considered the basis of information dengue vector prevention and control in Ternate City.

Potential application of Gustatory Receptor 1 (CmegGr1) gene as a molecular marker for identification of Chrysomya megacephala (Diptera: Calliphoridae)

@#Chrysomya megacephala larvae can easily be identified using cheap traditional microscopy techniques. Nevertheless, identification using taxonomy keys may be hampered, if the morphological characteristics of the larvae are incomplete, or immature for microscopic identification. To overcome the difficulty of species determination, molecular identification has gained relevance and is applied in forensic investigations. This study aimed to identify a novel target gene, known as the gustatory receptor 1 gene (CmegGr1), which has never been used for identification. The third instar larvae of Ch. megacephala (n = 30) and eight other forensically important fly species were obtained from two sources; rabbit carcasses and the Forensic Entomology Unit collection. Their DNAs were extracted and the CmegGr1 gene was amplified using polymerase chain reaction (PCR). The resulting sequences were subjected to phylogenetic analysis. A 209 bp fragment of the CmegGr1 gene was successfully amplified in 80% (24/30) of Ch. megacephala samples, while all of the non-Ch. megacephala species were not amplified. The phylogenetic analysis revealed that the evolutionary tree of CmegGr1 shares many traits with the 21a gustatory receptors of Calliphora stygia and Lucilia cuprina (Gr21a), which are also classified as necrophagous fly species. The high specificity of species identification was demonstrated in the present study using DNA barcoding, which led to the conclusion that the CmegGr1 gene could serve as an alternative marker for identifying Ch. megacephala.

Mosquito larva distribution and natural Wolbachia infection in campus areas of Nakhon Ratchasima, Thailand

Objective: To determine the prevalence of mosquito larvae in campus areas and the infection rate of endosymbiotic bacteria, Wolbachia in mosquito larvae. Method: The mosquito larvae samples were collected in residential areas and academic buildings of Suranaree University of Technology located in Northeastern Thailand during 2017-2018. Mosquito species identification was performed using GLOBE mosquito protocols and Rattanarithikul & Panthusiri's keys. The gene encoding for the surface protein of Wolbachia was amplified by PCR and confirmed by DNA sequencing. Results: Armigeres sp. is the highest proportion of mosquito larvae followed by Culex spp., Aedes albopictus, Aedes aegypti, and Toxorynchites spp., respectively. Aedes aegypti have breeding sites mostly in the containers found indoors, whereas the main breeding sites of Aedes albopictus were found in both outdoors and indoors. The House Index and Breteau Index for Aedes spp. was more than 5% and 20%, respectively, in both areas, indicating that these areas are dengue sensitive. The highest proportion of Wolbachia infection was found in the larvae of Culex spp. (86.21%), followed by Aedes albopictus (69.23%) and rarely detected in Aedes aegypti (9.09%). Conclusion: The present study reported the first natural infection of Wolbachia in mosquito larvae in Thailand. Our result suggested that the mosquito species containing higher proportion of Wolbachia are less likely to be vectors for dengue. Therefore, Wolbachia transfection in mosquito larvae could be applied as a biocontrol for dengue and other mosquito-borne disease prevention.

Notes on the biological development of the darkling beetle Blaps nefrauensis nefrauensis Seidlitz, 1893 (Coleoptera: Tenebrionidae) / Notas sobre o desenvolvimento biológico do besouro escuro Blaps nefrauensis nefrauensis Seidlitz, 1893 (Coleoptera: Tenebrionidae)

Several endemic species of Blaps occur in Tunisia, and the species Blaps nefrauensis nefrauensis has been reported in Moulares (urban zone in west-central Tunisia), where it lives and reproduces in home gardens and old buildings. The aim of this work is to study the life cycle of the darkling beetle, considering both field and laboratory rearing conditions. As a result, the beetle species has different developmental stages (egg, larva, prepupa, pupa, and adult) that last about 15 months. Each year during the same period, adults emerge (early summer) and expire (late autumn), larvae hatch (late summer) and pupate (early summer). There is only one generation per year. Females began laying eggs in late July. The eggs were ovoid, white, and about 2.7 mm in length and 1.5 mm in width. Embryogenesis took an average of nine days. The first instar larvae were at initially only 4.5 mm long and ivory white in color. A brief description of the newly egg hatched larva was provided; thus, the nerve fibers innervating the apical setae in the antennae and ligula were detected. Further light microscopic examination of the embryo before hatching from the egg pointed out that the antennal sensilla are protected during the embryogenesis stage.

Várias espécies endêmicas de Blaps ocorrem na Tunísia, e a espécie Blaps nefrauensis nefrauensis foi relatada em Moulares (zona urbana no centro-oeste da Tunísia), onde vive e se reproduz em jardins domésticos e prédios antigos. O objetivo deste trabalho é estudar o ciclo de vida do besouro escuro, considerando as condições de criação em campo e em laboratório. Como resultado, a espécie de besouro tem diferentes estágios de desenvolvimento (ovo, larva, prepupa, pupa e adulto) que duram cerca de 15 meses. Todos os anos, durante o mesmo período, os adultos emergem (início do verão) e expiram (final do outono), as larvas eclodem (final do verão) e se tornam pupas (início do verão). Existe apenas uma geração por ano. As fêmeas começaram a botar ovos no final de julho. Os ovos eram ovóides, brancos, com cerca de 2,7 mm de comprimento e 1,5 mm de largura. A embriogênese demorou em média nove dias. As larvas de primeiro instar tinham inicialmente apenas 4,5 mm de comprimento e uma cor branca marfim. Foi fornecida uma breve descrição da larva recém-eclodida; assim, as fibras nervosas que inervam as cerdas apicais nas antenas e ligulas foram detectadas. Um exame microscópico de luz posterior do embrião antes da eclosão do ovo mostrou que as sensilas antenais são protegidas durante o estágio de embriogênese.

Notes on the biological development of the darkling beetle Blaps nefrauensis nefrauensis Seidlitz, 1893 (Coleoptera: Tenebrionidae)

Abstract Several endemic species of Blaps occur in Tunisia, and the species Blaps nefrauensis nefrauensis has been reported in Moulares (urban zone in west-central Tunisia), where it lives and reproduces in home gardens and old buildings. The aim of this work is to study the life cycle of the darkling beetle, considering both field and laboratory rearing conditions. As a result, the beetle species has different developmental stages (egg, larva, prepupa, pupa, and adult) that last about 15 months. Each year during the same period, adults emerge (early summer) and expire (late autumn), larvae hatch (late summer) and pupate (early summer). There is only one generation per year. Females began laying eggs in late July. The eggs were ovoid, white, and about 2.7 mm in length and 1.5 mm in width. Embryogenesis took an average of nine days. The first instar larvae were at initially only 4.5 mm long and ivory white in color. A brief description of the newly egg hatched larva was provided; thus, the nerve fibers innervating the apical setae in the antennae and ligula were detected. Further light microscopic examination of the embryo before hatching from the egg pointed out that the antennal sensilla are protected during the embryogenesis stage.

Resumo Várias espécies endêmicas de Blaps ocorrem na Tunísia, e a espécie Blaps nefrauensis nefrauensis foi relatada em Moulares (zona urbana no centro-oeste da Tunísia), onde vive e se reproduz em jardins domésticos e prédios antigos. O objetivo deste trabalho é estudar o ciclo de vida do besouro escuro, considerando as condições de criação em campo e em laboratório. Como resultado, a espécie de besouro tem diferentes estágios de desenvolvimento (ovo, larva, prepupa, pupa e adulto) que duram cerca de 15 meses. Todos os anos, durante o mesmo período, os adultos emergem (início do verão) e expiram (final do outono), as larvas eclodem (final do verão) e se tornam pupas (início do verão). Existe apenas uma geração por ano. As fêmeas começaram a botar ovos no final de julho. Os ovos eram ovóides, brancos, com cerca de 2,7 mm de comprimento e 1,5 mm de largura. A embriogênese demorou em média nove dias. As larvas de primeiro instar tinham inicialmente apenas 4,5 mm de comprimento e uma cor branca marfim. Foi fornecida uma breve descrição da larva recém-eclodida; assim, as fibras nervosas que inervam as cerdas apicais nas antenas e ligulas foram detectadas. Um exame microscópico de luz posterior do embrião antes da eclosão do ovo mostrou que as sensilas antenais são protegidas durante o estágio de embriogênese.

Early development of two commercially valuable fish from the lower Amazon River, Brazil (Characiformes: Serrasalmidae)

We described the early development of Myloplus asterias and M. lobatus, two fish species of the Serrasalmidae using morphological, meristic, and morphometric characteristics. These herbivores serrasalmids are heavily fished because of their commercial importance in the Amazon. The individuals were collected between 2016 and 2020 in the limnetic zone of open water and macrophyte stands in the Lower Amazon River floodplain. We tested different growth models to identify the development pattern of these species. During the initial ontogeny, these two species can be differentiated mainly by the myomeres total number, 37 to 39 in M. asterias vs. 40 to 41 in M. lobatus, the pigmentation pattern, and dorsal-fin ray numbers. These characteristics are effective in distinguishing these species from other serrasalmids. The morphometric relationships were also different between these two species, showing distinct patterns in growth between the analyzed features. An identification key for larvae of some sympatric Serrasalmidae species from the Amazon basin is presented. Our expectation is that this study will contribute to the knowledge of the initial ontogeny and the biology of fish species in the Neotropical region.(AU)

Descrevemos o desenvolvimento inicial de Myloplus asterias e M. lobatus, duas espécies de peixes da família Serrasalmidae usando características morfológicas, merísticas e morfométricas. Esses serrasalmídeos herbívoros são fortemente pescados devido à sua importância comercial na Amazônia. Os indivíduos foram coletados entre 2016 e 2020 na zona limnética de águas abertas e em bancos de macrófitas na planície de inundação do baixo rio Amazonas. Testamos diferentes modelos de crescimento para identificar o padrão de desenvolvimento dessas espécies. Durante a ontogenia inicial, essas duas espécies podem ser diferenciadas pelo número total de miômeros; 37 a 39 em M. asterias vs. 40 a 41 em M. lobatus; pelo padrão de pigmentação e número de raios da nadadeira dorsal. Essas características são eficazes para distinguir essas espécies de outros serrasalmídeos. As relações morfométricas também foram diferentes entre as duas espécies, com distintos padrões de crescimento entre as características analisadas. Uma chave de identificação para larvas de algumas espécies simpátricas de Serrasalmidae da bacia amazônica é apresentada. Nossa expectativa é que este estudo contribua para o conhecimento da ontogenia inicial e da biologia das espécies de peixes da região Neotropical.(AU)