Single-Larva RNA Sequencing Identifies Markers of Copper Toxicity and Exposure in Early Mytilus californianus Larvae

One of the challenges facing efforts to generate molecular biomarkers for toxins is distinguishing between markers that are indicative of exposure and markers that provide evidence of the effects of toxicity. Phenotypic anchoring provides an approach to help segregate markers into these categories based on some phenotypic index of toxicity. Here we leveraged the mussel embryo-larval toxicity assay in which toxicity is estimated by the fraction of larvae that exhibit an abnormal morphology, to isolate subsets of larvae that were abnormal and thus showed evidence of copper-toxicity, versus others that while exposed to copper exhibited normal morphology. Mussel larvae reared under control conditions or in the presence of increasing levels of copper (3–15 μg/L Cu2+) were physically sorted according to whether their morphology was normal or abnormal, and then profiled using RNAseq. Supervised differential expression analysis identified sets of genes whose differential expression was specific to the pools of abnormal larvae versus normal larvae, providing putative markers of copper toxicity versus exposure. Markers of copper exposure and copper-induced abnormality were involved in many of the same pathways, including development, shell formation, cell adhesion, and oxidative stress, yet unique markers were detected in each gene set. Markers of effect appeared to be more resolving between phenotypes at the lower copper concentration, while markers of exposure were informative at both copper concentrations.

ANALYSIS OF MANAGERIAL COMPONENTS IN MOSQUITO VECTORS (AEDES AEGYPTI) CONTROL IN THE BUFFER AREA OF THE CLASS 1 SURABAYA PORT HEALTH OFFICE

ABSTRACTThe Aedes aegypti index in the buffer area of the Class 1 Surabaya Port Health Office was in accordance with the Standard Operating Procedures (SOP) as stated in the Regulation of the Indonesian Ministry of Health Number 431 of 2007 (431/MENKES/SK/IV/2007) on Technical Guidelines for Control of Environmental Health Risks in Ports/Airports/Cross-Border in the Context of Health Quarantine. The Port Health Office has responsibilities to control environmental risks, one of which was the Aedes aegypti mosquitos in the borders. This study was descriptive observational and aimed to analyze the managerial components in Aedes aegypti mosquito control in the buffer area ofthe Class 1 Surabaya Port Health Office in Surabaya. The research variables (factors that determine control activities) were manpower, money, methods, materials, markets, machines, and information. The research subjects were two officers at Class 1 Surabaya Port Health Office and five cadres who monitored larva. Data were analyzed descriptively. The results explained that manpower, money, materials, market, technology, and information were all in accordance with the SOP of the Surabaya Port Health Office. However, the larva survey method did not comply with the SOP as officers still used the visual method. It was concluded that managerial components in Aedes aegypti control by Class 1 Surabaya Port Health Office followed the SOP except in their larva survey method. The researchers recommended carrying out a larva survey method by taking 1 larva from each container (single larva) at a time. Keywords: Aedes aegypti, control vector, managerial components.

Assessment of Mosquito Collection Methods for Dengue Surveillance

Several methods exist to collect and assess the abundance of dengue vector mosquitoes, i.e., morning adult collection, pupal collection, ovitraps, human landing, and larval collection. Several of these methods are officially implemented to monitor mosquito density and make decisions on treatments for dengue control. This monitoring is also constrained by the need to conduct this assessment on a “one point/one day” process, meaning that once the threshold of 100 households is reached, the assessment is made, and the collectors teams move to another place, thus preventing the use of long-term sampling methods. This diversity of methods might be a source of variability and lack of statistical significance. There is also a lack of published data regarding the efficacy of these methods. Furthermore, the Stegomyia indices are shown to be not reliable for assessing the risk of dengue outbreaks. A mosquito survey was, thus, conducted in 39 locations corresponding to 15 dengue endemic provinces in Indonesia by using the different adult and larval collection methods recommended nationwide. A total of 44,675 mosquitoes were collected. The single larva method was the most efficient. Out of a total of 89 dengue-positive pools, the most frequently encountered virus was DENV2, which made up half of the positive samples, followed by DENV3 and DENV1, respectively. Factor analysis of mixed data showed that no correlation could be found between any methods and the presence of dengue virus in mosquitoes. Moreover, no correlation could be found between any methods and the incidence of dengue. There was no consistency in the efficacy of a given method from one site to another. There was no correlation between any of the parameters considered, i.e., method, incidence of dengue, location, and the presence of dengue virus in mosquitoes.

A New, Cheap, Easy to Use, Foldable and Portable Autocidal Ovitrap for Aedes Control at Community Level

Except for Yellow Fever there is still no operational vaccine, the Dengue vaccine (CYD-TDV, sold under the brand name Dengvaxia©) raised several issues and vaccination campaign were stopped. On the other hand there is still no specific treatment for the main arbovirus transmitted by Aedes aegypti and Aedes albopictus such as dengue, Chikungunya, Zika and recent outbreaks occurred all over the World. For WHO “vector control is the key strategy to control or prevent the transmission of dengue (and other arbovirus) but clearly there is a need for improved new vector control approaches. Only another one vaccine is available against arbovirus is against Japanese Encephalitis transmitted by Culex tritaeniorhynchus. In line with this recommended approaches of new tools to be used at community level we successfully developed and implemented in Angola a “Very Simple Ovitrap Model” which actually “collected” eggs of Aedes aegypti warning for risk of arbovirus outbreaks, which, unfortunately, occurred few years later. These ovitraps were transformed as a tool for vector control in adding few drops of Bacillus thuringiensis (Bt) in the water and several hundred of tests showed that not a single larva of Aedes aegypti was ever found in these “Bt fitted simple ovitrap”. To solve some operational issues (transport) we tested in Bali during 3 years several size and color of foldable container prepared as ovitraps and put in a garden among grass, banana trees etc, we thus collected eggs of Aedes albopictus, important vector of dengue and dengue hemorrhagic fever (DHF), a serious threat in Asia, and preliminary tests (unpub.) showed some resistance to insecticide increasing the need for control. According to these trials, made in complete field conditions, we proposed a new foldable model of ovitrap as a vector control tool presented as a kit to be used at community level with a 3 steps action: open it/fill it/use it.

Indeks Maya dan Indeks Entomologi Vektor Demam Berdarah Dengue di Kabupaten Mamuju Tengah, Sulawesi Barat

The existence of containers as a potential habitat breeding places for mosquitoes in the community can affect the population density of Aedes. The maya index is an indicator to measure the number of containers that can be a breeding places for mosquitoes. This study aims to determine maya index and entomology index in Central Mamuju Regency, West Sulawesi Province. This study was an observational study conducted in 2015 in three sub-districts (Topoyo, Tappilina, and Karossa) in Central Mamuju Regency. Data collection through structured interviews with household heads in 100 randomly selected houses using the closest house method and measuring larvae density using the single larva method. The results showed that the Breeding Risk Index (BRI) for the three sub-districts in Central Mamuju Regency are mostly in the medium category, while Hygiene Risk Index (HRI) and Maya Index (MI) are in a low category. Based on House Index (HI) and Container Index (CI) indicators means have a moderate risk of transmission Dengue.

Redescription of the tadpole of rare Giant Torrent Frog Megaelosia boticariana Giaretta & Aguiar, 1998 (Anura, Hylodidae) with comments on natural history

The Hylodidae genus Megaelosia Miranda-Ribeiro comprises seven frog species that inhabit the Atlantic Rainforest in Southeastern Brazil (Da Silva et al. 2018). Larvae and adults of Hylodidae are commonly associated with lotic streams (Giaretta et al. 1993; Silva-Soares et al. 2015). Tadpoles of all Megaelosia species but Megaelosia bocainensis Giaretta, Bokemann & Haddad are formally described, even though many of these descriptions are restricted to few lines and several anatomical details were overlooked (e.g., Megaelosia lutzae; Izecksohn & Gouvêa 1985). The tadpole of M. boticariana was briefly described by Giaretta & Aguiar (1998) based on a single larva with no measurements or figures. Currently, M. boticariana is only known from its type locality and São Francisco Xavier, both in São Paulo state portion of the Serra da Mantiqueira, Brazil (Muscat et al. 2020). The lack of information, coupled with the rarity of the species, reinforces the importance of taxonomic and natural history data to subside works on other fields. In this context, we present a complete redescription of the external anatomy of the tadpoles of M. boticariana and comment on their natural history.

Review of Oculogaster Kluge 2016 (Ephemeroptera, Baetidae, Procloeon Bengtsson 1915)

The subgenus Oculogaster Kluge 2016 is divided into two species groups: the «African group» includes Procloeon (Oculogaster) cylindroculum Kimmins 1956, P. (O.) barnardi sp. n., P. (O.) niger sp. n., unnamed species P. (O.) sp. «Wolfkloof» and presumably P. (O.) silvicola Gillies 1997; the «Asian group» includes P. (O.) album Kluge 2016, P. (O.) regularum Müller-Liebenau & Hubbard 1985, P. (O.) malabarensis sp. n., presumably Procloeon julia (Gillies 1949) comb. n. and unnamed species P. sp. «Thailand-3», P. sp. «Sulawesi» and P. sp. «Philippines». New South African species P. (O.) barnardi sp. n. and P. (O.) niger sp. n. are described from South Africa based on larvae, subimagines and imagines of both sexes associated by rearing; among them, P. (O.) barnardi sp. n. was formerly described by Barnard (1932) under the name «Austrocloeon africanum», being wrongly regarded to be the species originally described as Cloeon africanum Esben-Petersen 1913. Characters of larva and female subimago of another South African species, P. (O.) sp. «Wolfkloof» are described based on a single specimen. Two closely related species, P. (O.) regularum Müller-Liebenau & Hubbard 1985 from Sri Lanka and P. (O.) malabarensis sp. n. from India are described based on larvae, subimagines and imagines of both sexes associated by rearing; these two species have 2 (rarely 1 or 3) cross veins in pterostigma, in contrast to other species of Oculogaster which normally have one vein (rarely two veins). Procloeon julia is presumptively placed in Oculogaster based on the presence of one vein in pterostigma. Unnamed species P. sp. «Thailand-3», P. sp. «Sulawesi» and P. sp. «Philippines» are presumably placed in Oculogaster; each is briefly described based on a single larva.

Potensi Transmisi Demam Berdarah Dengue Berdasarkan Indeks Entomologi dan Maya Indeks di Tiga Kelurahan Kecamatan Sukajadi Kota Pekanbaru

Latar belakang: Demam berdarah dengue (DBD) merupakan penyakit yang masih menjadi masalah kesehatan di Indonesia yang disebabkan oleh virus dengue dan ditularkan melalui nyamuk Aedes aegypti. Kejadian DBD selalu ada setiap tahun di Pekanbaru dari tahun 2014-2016 terjadi peningkatan kasus, 2017-2018 kasus DBD mulai menurun tetapi diawal tahun 2019 kasus DBD kembali terjadi peningkatan dan sudah ada kematian sehingga memerlukan perhatian yang lebih serius untuk meningkatkan kewaspadaan terhadap penyakit tersebut. Tujuan penelitian ini adalah untuk menganalisis potensi transmisi DBD berdasarkan indeks entomologi dan maya indeks di tiga kelurahan Kecamatan Sukajadi yang merupakan daerah endemis DBD. Metode: Penelitian survei larva Aedes aegypti dengan pendekatan cross sectional yang dilakukan pada bulan Juli 2019 di tiga kelurahan yaitu Kelurahan Kampung Tengah, Kampung Melayu, Kedungsari Kecamatan Sukajadi. Identifikasi larva menggunakan metode single larva method. Hasil: Sebanyak 181 rumah yang disurvey, didapatkan 822 kontainer yang terdiri dari 683 controllable sites dan 139 disposable sites. Angka bebas jentik sebesar 89,5%, container index 3,4%, house index 10,5% dan Breteau index 21%. Status Maya indeks (MI) yang diukur berdasarkan breeding risk index dan hygiene risk index didapatkan sebanyak 55,80% rumah termasuk dalam status sedang dan 15,47% termasuk ke dalam status MI tinggi. Sebaran keberadaan larva Aedes aegypti berdasarkan letak geografis cukup merata di Kelurahan Kampung Melayu dan Kampung Tengah.Simpulan: Berdasarkan hasil indeks entomologi tersebut didapatkan density figure dalam kategori sedang yang artinya wilayah ini mempunyai potensi transmisi sedang untuk kejadian penyakit DBD. ABSTRACT Title : Transmission Potential of Dengue hemorrhagic fever based on entomology index and maya index in three sub-districts, Sukajadi District, Pekanbaru CityBackground: Dengue hemorrhagic fever (DHF) is a disease still a health problem in Indonesia caused by the dengue virus that is transmitted through Aedes aegypti. The incidence of DHF is always there every year in Pekanbaru form 2014-2016 there was an increase in cases, from 2017-2018 dengue cases began to decline, but in early 2019 dengue cases increased again and there have been deaths so it requires more serious attention.. The objectives of this study was to analyze the potential transmission of DHF based on the entomology index and maya index in three sub-districts of Sukajadi district.Method: The research was a survey of larva Ae.aegypti in July 2019 from 181 houses in the three village of Sukajadi District. Larvae identification using the single larvae method.Result: A total of 181 houses surveyed obtained 822 containers consisting of 683 controllable sites and 139 disposable sites. The larvae free rates is 89.5%, container index 3.4%, house index 10,5% and Breteau index 21%. Maya index is measures based on breeding risk index and hygiene risk index shows that there are 55.8% of houses in the area are classified as moderate and 15.47% are included in high. The distribution of the existence of Ae.aegypti larvae based on the geographical location is quite evenly distributed in Kampung Melayu and Kampung Tengah.Conclusion: It is found that density figure is in the medium, which means that region has moderate transmission potential for the incidence of DHF

Profil Indeks Larva Pada Delapan Wilayah Puskesmas di Surabaya

<p><strong>ABSTRACT</strong></p><p><strong>Background : </strong>Dengue virus infection is still a health problem in Surabaya. On 2015 Insidence Rate (IR) for Surabaya is 22.50 and the Case Fatality Rate (CFR) is 2%. This figure has decreased compared to 2014, but has not met the target achievement indicator for the P2DBD target CFR &lt;1%.</p><p><strong>Method : </strong>The study was conducted in the rainy season, May 2015 in several Puskesmas areas in Surabaya. Samples were randomly selected as many as 378 houses spread in eight health centers (Pegirian, Krembangan Selatan, Hamlet Wall, Tambak Rejo, Putat Jaya, Simomulyo, Sawahan and Tanahkali Kedinding). The larvae survey was conducted using the single larva method. Larvae were taken from breeding sites and stored in specimen bottles for species identification at the Parasitology Laboratory Faculty of Medicine Universitas Airlangga.</p><p><strong>Result :</strong> The highest House Index (HI) was obtained (60.46%) or the 39.54% larvae free rate in Simomulyo and the lowest HI (33.33%) or 66.67% ABJ in Pegirian Puskesmas. Larvae species identification showed 100% was Aedes aegypti. The high HI value at the Simomulyo Health Center is not in line with the number of cases of dengue virus infection in 2015 which was 4 cases. Whereas in Pegirian Health Center with the lowest HI value found 11 cases.</p><p><strong>Discussion :</strong> These results could be due to the possibility because the study did not calculate larval density and high population mobility in the area.</p><p><strong>Conclusion</strong>: House Index values are linier with container index but not linear with the number of cases of dengue virus infection</p><p><strong>Keywords:</strong> House Index, larvae free rate, dengue virus infection, single larva method</p>

Penentuan Angka Kepadatan (Density Figure) dan Angka Bebas Jentik (ABJ) Larva Aedes aegypti di Rw 02, Kelurahan Margahayu, Kecamatan Bekasi Timur, Kota Bekasi, Jawa Barat

World Health Organization (WHO) memperkirakan ada 390 juta infeksi dengue/tahun. Pada tahun 2016, di Indonesia tercatat sebanyak 204.171 kasus demam berdarah dengue (DBD) dengan jumlah kematian 1.598 orang. Jawa Barat merupakan provinsi dengan kasus DBD terbanyak. Di RW 02, Kelurahan Margahayu, Kecamatan Bekasi Timur, Kota Bekasi, Jawa Barat terdapat warga yang terkena DBD. Selain itu, kurangnya kegiatan 3M-Plus dan tidak adanya kegiatan kader juru pemantau jentik (jumantik) meningkatkan potensi penularan penyakit DBD di tempat tersebut. Tujuan penelitian ini adalah untuk menentukan tingkat kepadatan larva Aedes aegyptidan menentukan tingkat pengetahuan, sikap dan perilaku warga mengenai tempat penampungan air (TPA) dan 3M-Plus berdasarkan ada/tidaknya larva Ae. aegypti pada rumah warga, serta hubungan antara keduanya. Penelitian ini bersifat deskriptif analitik dengan pendekatan cross sectional. Seratus sampel rumah dipilih secara random sampling. Spesimen diambil dengan metode single larva. Data disajikan dalam tabel distribusi frekuensi dan dianalisis dengan uji chi-square. Hasil penelitian menunjukkan Angka Kepadatan (Density Figure) = 6 (House Index = 38%, Container Index = 22%, Breteau Index = 50), sehingga kepadatan larvanya tergolong tinggi. Angka Bebas Jentik (ABJ) sebesar 62%, yang berarti belum memenuhi target ABJ ≥ 95%. Tingkat pengetahuan, sikap dan perilaku warga yang rumahnya tidak terdapat larva Ae. aegyptitergolong baik dan berlaku sebaliknya. Terdapat hubugan antara keberadaan larva Ae. aegypti dengan pengetahuan (p= 0,022), sikap (p= 0,028) dan perilaku (p= 0,000) warga mengenai TPA dan kegiatan 3M-Plus. Berdasarkan hasil tersebut, disarankan agar warga lebih giat melakukan kegiatan 3M-Plus dan mengaktifkan kegiatan kader jumantik di RW 02.Kata Kunci :Aedes aegypti, Density Figure, Angka Bebas Jentik