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Proclotting enzymes (PCE) are a serine protease-containing CLIP domain and play roles in development of embryonic and innate immune response. Little is known about PCE in mosquitoes, but only three melanization regulatory modules that involve proteases with the CLIP domain have been reported. We cloned the PCE gene and reported the nucleotide sequence encoding a proclotting enzyme in Aedes aegypti L. We did in-silico protein translation, alignment with proclotting proteinase sequences of the other species of different taxonomic orders, and phylogenetic analysis where PCE in Ae. aegypti fit the hypotheses of positive or adaptive evolution. The fundamental information could be applied in strategies for biological control of medically important insects. [ABSTRACT FROM AUTHOR]

In prokaryotes and eukaryotes, the Period (PER) gene is important for the circadian clock that regulates some rhythmic biochemical, physiological, and behavioral processes. PER messenger amplification was done by RT-PCR from southern house mosquito, Culex quinquefasciatus Say, larvae collected at three locations in the State of Nuevo Leon, northeastern Mexico. The amplified products were cloned, sequenced, and compared/analyzed with those of other species of mosquitoes. A 3,093-bp amplified product was synthesized by PCR, which is the complete coding DNA sequence (3,027 bp) of PER encoding an open reading frame of 996 amino acid residues and much smaller than the orthologous in other mosquitoes analyzed. No other simultaneous amplification was visualized; thus, possible alternative splicings were discarded or at least not detected by the method. Because PER is a widely conserved gene in prokaryotes and eukaryotes and maintaining colonies of southern house mosquitoes is relativity easy, determining the sequence of the gene in the mosquito provides the possibility of using the gene as a model and generating data on the role in the circadian clock. [ABSTRACT FROM AUTHOR]

In eukaryotes, the timeless (TIM) gene plays a fundamental role in the control of the circadian clock by regulating several biochemical, physiological, and behavioral processes. RT-PCR was used to amplify TIM mRNA from larval southern house mosquito, Culex quinquefasciatus Say, from three locations in the State of Nuevo Leon in northeastern Mexico. The 3062 bp TIM cDNA was assembled from three overlapping PCRs that were cloned and sequenced. DNA sequence analysis used amplified cDNA fragments to complete coding the DNA sequence (2991 bp). Tim ORF consisting of 996 codons was shorter than the orthologous genes from other mosquito species. A series of deletions in the core sequence of TIM were detected. No other amplification products were visualized, and thus, the possibility of alternative spliced mRNA species was discarded. Because TIM is a widely conserved gene in eukaryotes, and maintaining colonies of Cx. quinquefasciatus is easy, determining the sequence of the TIM gene in the mosquito provides the possibility of its use as a model for circadian clock experimentation. [ABSTRACT FROM AUTHOR]

Bluetongue virus is the etiological agent of a disease characterized by bleeding in domestic and wild ruminants. Different viral strains can infect a ruminant species and severity of infection is variable. Transmission of the virus under natural conditions is through the bite of a mosquito of the genus Culicoides (Ceratopogonidae), but also can be transmitted by infected semen or embryos. The genus Culicoides is a vector of other diseases such as African horse sickness. Specimens are about 0.5 to 5 mm in size. There are about 1,400 species of Culicoides, only 32 of which can act as biological vectors. This is because of the receiver with the intestinal cells where amplification occurs in viral load. A segment 10 of the viral genome from 16 viral serotypes (of approximately 26 serotypes reported) was used. The sequences were aligned, and the consensus sequence was obtained. Subsequently, a pair of oligonucleotides was designed on a highly conserved region that theoretically allows co-amplification of the strains. Sites for restriction enzymes were identified to create a specific electrophoretic pattern for each serotype. It was possible to theoretically amplify a gene segment of 16 bluetongue virus serotypes with one set of oligonucleotides and the serotypes can be differentiated with two restriction enzymes. The proposed method could become a simple, inexpensive, and practical routine tool for type-specific identification of bluetongue virus. [ABSTRACT FROM AUTHOR]

Aedesaegypti L. is the most important vector of arboviruses such as dengue, Zika, chikungunya, Mayaro, and yellow fever, which impact millions of people's health per year. MicroRNA profile has been described in some mosquito species as being important for biological processes such as digestion of blood, oviposition, sexual differentiation, insecticide resistance, and pathogens dissemination. We identified the miRNAs of Ae.aegypti females, males and eggs of a reference insecticide susceptible strain New Orleans and compared them with those other insects to determine miRNA fingerprint by new-generation sequencing. The sequences were analyzed using data mining tools and categorization, followed by differential expression analysis and conservation with other insects. A total of 55 conserved miRNAs were identified, of which 34 were of holometabolous insects and 21 shared with hemimetabolous insects. Of these miRNAs, 32 had differential expression within the stages analyzed. Three predominant functions of miRNA were related to embryonic development regulation, metamorphosis, and basal functions. The findings of this research describe new information on Ae.aegypti physiology which could be useful for the development of new control strategies, particularly in mosquito development and metamorphosis processes. [ABSTRACT FROM AUTHOR]

Introduction: Ursus americanus Pallas 1780 is the largest carnivore and the only ursid in Mexico. It is considered an endangered species in the country because its distribution and population have been reduced by up to 80% because of habitat loss or furtive hunting. These problems can lead to a diet change, which could result in metabolic disorders, such as fatty acid β-oxidation defects or organic acid metabolism disorders. In our study, a free amino acid and acylcarnitine profile was characterized. Methods: Peripheral blood samples were drawn from nine free-ranging black bears in a period of five months, from June to October of 2019 in Northeastern Mexico, and 12 amino acids and 30 acylcarnitines were determined and quantified. Age differences were observed in the samples through ANOVA and post-hoc Tukey test. Results: Only three metabolites showed a significant difference with age: alanine (Ala) [cubs vs juvenile], free-carnitine (C0) [juvenile vs cubs] and acetylcarnitine (C2) [cubs vs adults and juvenile vs cubs]. Conclusion: Metabolites with variability due to age were identified, making them potential biomarkers to monitor metabolic status as early diagnosis in endangered species. This is the first study of black bear amino acid and acylcarnitine profiles, and the values found could be used as reference for free amino acid and acylcarnitine concentrations in further studies of the species. [ABSTRACT FROM AUTHOR]

Megalopyge opercularis (Smith) (Lepidoptera: Megalopyridae) is a nocturnal moth of medical importance because it causes adverse immunological reactions in humans. In this study, we determined the microbiota composition of M. opercularis at the larval (caterpillars) and adult (moths) stages by next-generation sequencing. DNA was extracted from the caterpillars and moths, and the 16S rRNA prokaryote gene was then amplified and sequenced with next-generation sequencing to assess bacterial richness. Comparison of the microbiota of the caterpillars and adults revealed variation in species composition and diversity. The microbiota of the caterpillars of M. opercularis was composed of 259 species, dominated by the families Geodermatophilaceae (12%), Propionibacteriaceae (10.41%), Clostridiaceae (9.63%), and Nitriliruptoraceae (7.72%). In the adult moths, we found 138 species, and the most abundant families were Nostocaceae (24%) and Methylobacteriaceae (21%). Species richness in M. opercularis was higher in the caterpillars compared to adults. We determined that only some groups of bacteria could persist from 1 stage to another. The results obtained are essential to know about the ecology of M. opercularis and contribute to our understanding of the impact that microorganisms have on the physiological mechanisms of poisonous lepidopterans. Megalopyge opercularis (Smith) (Lepidoptera: Megalopyridae) es una especie de polilla nocturna perteneciente a la familia Megalopyridae de importancia médica debido a las reacciones inmunológicas causadas en humanos por sustancias urticantes ubicadas en sus estructuras espinosas. En este estudio, determinamos el microbioma de M. opercularis en su estado larval y en su estado adulto. Se extrajo el ADN de larvas y adultos y se amplificó el gen rRNA 16S mediante tecnologías de secuenciación de nueva generación. Las comparaciones del microbioma larval contra el microbioma adulto revelaron variación tanto en la composición como en la diversidad de especie. El microbioma larval de M. opercularis está compuesto de 259 especies, dominado por las familias Geodermatophilaceae (12%), Propionibacteriaceae (10.41%), Clostridiaceae (9.63%), y Nitriliruptoraceae (7.72%). En el microbioma adulto encontramos 138 espcies, las familias más abundantes fueron Nostocaceae (24%) y Methylobacteriaceae (21%). La riqueza de especies encontradas en M. opercularis fue mayor en las orugas en comparación con los adultos. Determinamos que solo algunos grupos de bacterias podían persistir de una etapa a otra. Los resultados obtenidos son importantes para conocer la ecología de esta polilla y contribuyen a comprender el impacto que tienen los microorganismos en los mecanismos fisiológicos de los lepidópteros venenosos. [ABSTRACT FROM AUTHOR]

Melanaphis sacchari (Zehntner) is a pest of agricultural importance because of its ability to destroy crops of Sorghum bicolor (L.) Moench. Association with endosymbiotic bacteria affect aphid fitness and are influenced by such aspects as geography and association with the host plant. Composition of microbiota of M. sacchari was determined by extracting DNA from the gut of adult individuals (n = 60) and sequencing the 16s rRNA gene by next-generation sequencing (Ion TorrentTM). The most abundant phylum was Proteobacteria (98.88%), and the dominant genera were Leclercia (25.28%), Pantoea (16.54%), Trabulsiella (9.06%), and Serratia (2.94%). These results can be use to study the importance of primary symbiotic bacteria of M. sacchari and their relationship with agricultural production. [ABSTRACT FROM AUTHOR]

The southern house mosquito, Culex quinquefasciatus Say (Diptera: Culicidae), transmits pathogens and is one of the most important vectors of human disease. The carboxylesterase mosquito genes encoded esterase enzyme involved in the mechanism that provides resistance to insecticides. We amplified by RTPCR four alternative transcripts from the esterase gene not reported previously (A, B, C, and D) from Cx. quinquefasciatus larvae from Monterrey, N.L., northeastern Mexico. Through cDNA sequences, we predicted amino acid sequences used to model the three-dimensional structure of protein isoforms. We determined dN and dS ratios to identify evolutionary forces which lead the family. PCR products were obtained from 5A esterases (1839, 1898, 1894, and 1953 bp). The translation sequence of the PCR products showed polypeptides of 612, 473, 282, and 282 amino acids in length for isoforms A, B, C, and D, respectively. Three exons and two introns were obtained in the analyzed sequences. The study sequences were compared with those previously published showing 98.7, 98.5, 91.5, and 91.5% amino acid similarity for isoforms A, B, C, and D, respectively. The threedimensional models showed that only ORF-D had the three amino acids corresponding to the catalytic triad, while ORF-A and ORF-C lacked one or two of the three amino acids. Esterase genes are under purifying of selection; a clue the members are functional. The processes might be useful in monitoring evolutionary dynamics of known carboxylesterase genes and identifying new carboxylesterase splicing. [ABSTRACT FROM AUTHOR]

Sepik virus, a pathogen transmitted by arthropods to humans is in the flavivirus family. For this reason, the genomic study of the Sepik virus is relevant in vector-borne diseases field. Genomic mining of the virus and comparison of its coding regions were done to identify sequences that represent greater and lesser conservation than other phylogenetically correlated viruses and to identify regions of greater selection pressure within its genome. The Sepik genome was mapped and analyzed by alignment processes. Selection pressure was estimated using the Pamilio-Banachi-Li method. The genome of the Sepik virus is a 17-motif polyprotein. Within these motifs, the codifying regions of greatest conservation were immunoglobulin-like domain III (82.77%) and the DEAD domain (68.42%), while some domains indicated selection pressure. The selection pressure predicted for certain domains suggested evolutionary pressure on the virus that represented medical impact on virulence and biological risk of the virus. [ABSTRACT FROM AUTHOR]

Culex quinquefasciatus Say is a vector of many pathogens of humans, and both domestic and wild animals. Personal protection, reduction of larval habitats, and chemical control are the best ways to reduce mosquito bites and, therefore, the transmission of mosquito-borne pathogens. Currently, to reduce the risk of transmission, the pyrethroids, and other insecticide groups have been extensively used to control both larvae and adult mosquitoes. In this context, amino acids and acylcarnitines have never been associated with insecticide exposure and or insecticide resistance. It has been suggested that changes in acylcarnitines and amino acids profiles could be a powerful diagnostic tool for metabolic alterations. Monitoring these changes could help to better understand the mechanisms involved in insecticide resistance, complementing the strategies for managing this phenomenon in the integrated resistance management. The purpose of the study was to determine the amino acids and acylcarnitines profiles in larvae of Cx. quinquefasciatus after the exposure to different insecticides. Bioassays were performed on Cx. quinquefasciatus larvae exposed to the diagnostic doses (DD) of the insecticides chlorpyrifos (0.001 μg/mL), temephos (0.002 μg/mL) and permethrin (0.01 μg/mL). In each sample, we analyzed the profile of 12 amino acids and 31 acylcarnitines by LC-MS/MS. A t-test was used to determine statistically significant differences between groups and corrections of q-values. Results indicates three changes, the amino acids arginine (ARG), free carnitine (C0) and acetyl-carnitine (C2) that could be involved in energy production and insecticide detoxification. We confirmed that concentrations of amino acids and acylcarnitines in Cx. quinquefasciatus vary with respect to different insecticides. The information generated contributes to understand the possible mechanisms and metabolic changes occurring during insecticide exposure. [ABSTRACT FROM AUTHOR]

Alkhurma hemorrhagic fever virus was first isolated at Jeddah, Saudi Arabia, in the early 1990s, and identified as a member of the Flaviviridae family. The virus has been confirmed only in the sand tampan tick, Ornithodoros savignyi (Audouin), and camel tick, Hyalomma dromedarii Koch. Symptoms of infection include headache, joint and muscle pain, vomiting, and thrombocytopenia, leading to hemorrhagic fever, which can cause death. The purpose of this article was to make a genomic-mined Alkhurma hemorrhagic fever virus from the genomic sequences previously reported in GenBank of NCBI. Genomes were divided into peptides, and different evolutionary parameters were derived using bioinformatics tools. The individual behavior of each of the peptides encoded in the genome of Alkhurma hemorrhagic fever virus is specific; the rate of change indicates different kinds of evolutionary pressure. Complete genome and NS3 protein are under positive selection (dN > dS, p < 0.05), while purifying selection is the main force that drives the evolution of envelope and NS5 proteins (dN < dS, p < 0.05). The spread of viruses such as Alkhurma hemorrhagic fever virus can be catastrophic if they affect the global population without antibodies. Genomic mining of Alkhurma hemorrhagic fever virus provides a powerful tool to design new strategies focused on variable regions of the genome in case of spread. [ABSTRACT FROM AUTHOR]