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Many compounds being considered as candidates for advanced biofuels are toxic to microorganisms. This introduces an undesirable trade‐off when engineering metabolic pathways for biofuel production because the engineered microbes must balance production against survival. Cellular export systems, such as efflux pumps, provide a direct mechanism for reducing biofuel toxicity. To identify novel biofuel pumps, we used bioinformatics to generate a list of all efflux pumps from sequenced bacterial genomes and prioritized a subset of targets for cloning. The resulting library of 43 pumps was heterologously expressed in Escherichia coli , where we tested it against seven representative biofuels. By using a competitive growth assay, we efficiently distinguished pumps that improved survival. For two of the fuels ( n ‐butanol and isopentanol), none of the pumps improved tolerance. For all other fuels, we identified pumps that restored growth in the presence of biofuel. We then tested a beneficial pump directly in a production strain and demonstrated that it improved biofuel yields. Our findings introduce new tools for engineering production strains and utilize the increasingly large database of sequenced genomes. Biofuels can be produced by microbes, but biofuel toxicity is a major obstacle to efficient production. Here, the authors identify efflux pumps that can effectively export biofuels, improving cell viability and increasing biofuel yields.

Xylan is a major component of the plant cell wall and the most abundant noncellulosic component in the secondary cell walls that constitute the largest part of plant biomass. Dicot glucuronoxylan consists of a linear backbone of β(1,4)-linked xylose residues substituted with α(1,2)-linked glucuronic acid (GlcA). Although several genes have been implicated in xylan synthesis through mutant analyses, the biochemical mechanisms responsible for synthesizing xylan are largely unknown. Here, we show evidence for biochemical activity of GUX1 (for GlcA substitution of xylan 1), a member of Glycosyltransferase Family 8 in Arabidopsis (Arabidopsis thaliana) that is responsible for adding the glucuronosyl substitutions onto the xylan backbone. GUX1 has characteristics typical of Golgi-localized glycosyltransferases and a K m for UDP-GlcA of 165 μM. GUX1 strongly favors xylohexaose as an acceptor over shorter xylooligosaccharides, and with xylohexaose as an acceptor, GlcA is almost exclusively added to the fifth xylose residue from the nonreducing end. We also show that several related proteins, GUX2 to GUX5 and Plant Glycogenin-like Starch Initiation Protein6, are Golgi localized and that only two of these proteins, GUX2 and GUX4, have activity as xylan α-glucuronosyltransferases.

Abstract A study was conducted to evaluate the effect of Piper nigrum (black pepper) leaf extract on on the growth performance, proximate composition, hematological parameters, and immune response of Labeo rohita fingerlings with an average weight of 22.14 ± 0.98g. Aftrer acclimation for two weeks, fish (n=25) were randomly selected and placed in four glass aquaria (T0, T1, T2 and T3) at constant water temperature (30.0 ± 1.0 °C), pH (7.50 ± 0.5) and total hardness (200 ± 2.0 mgL,-1) for a period of 12 weeks, with three replicates each. Fish were fed with P. nigrum leaf extract supplemented feed @ 0.0%, 1.0%, 2.0% and 3.0% in T0, T1,T2 and T3, respectively. At the end of experiment, five fish were randomly selected from each aquaria for proximate composition, gut and skin microbial load, hematological parameters. Total proteins, albumins, and globulins were also recorded to evaluate immunological memory. The result revealed that fish in T2 showed better growth performance with an average weight gain of 56.11 ± 0.51 g. Thus, it had been concluded that Piper nigrum, a medicinal plant, can also be used to enhance the growth performance and immune response of Labeo rohita as attractive alternatives against antibiotics and vaccines and has shown no negative side effects on fish health as well as on its environment. Resumo Um estudo foi conduzido para avaliar o efeito do extrato da folha de Piper nigrum (pimenta-do-reino) sobre o desempenho de crescimento, composição centesimal, parâmetros hematológicos e resposta imune de alevinos de Labeo rohita com peso médio de 22,14 ± 0,98g. Após aclimatação por duas semanas, os peixes (n = 25) foram selecionados aleatoriamente e colocados em quatro aquários de vidro (T0, T1, T2 e T3) em temperatura constante da água (30,0 ± 1,0 °C), pH (7,50 ± 0,5) e dureza total (200 ± 2,0 mgL-1) por um período de 12 semanas, com três repetições cada. Os peixes foram alimentados com ração suplementada com extrato de folha de P. nigrum @ 0,0%, 1,0%, 2,0% e 3,0% em T0, T1, T2 e T3, respectivamente. Ao final do experimento, cinco peixes foram selecionados aleatoriamente de cada aquário para composição centesimal, carga microbiana intestinal e cutânea e parâmetros hematológicos. Proteínas totais, albuminas e globulinas também foram registradas para avaliar a memória imunológica. O resultado revelou que os peixes em T2 apresentaram melhor desempenho de crescimento com ganho de peso médio de 56,11 ± 0,51 g. Assim, concluiu-se que Piper nigrum, uma planta medicinal, também pode ser usado para melhorar o desempenho de crescimento e resposta imunológica de Labeo rohita como alternativas atraentes contra antibióticos e vacinas e não mostrou efeitos colaterais negativos na saúde dos peixes, bem como sobre seu ambiente.

The plant Golgi plays a pivotal role in the biosynthesis of cell wall matrix polysaccharides, protein glycosylation, and vesicle trafficking. Golgi-localized proteins have become prospective targets for reengineering cell wall biosynthetic pathways for the efficient production of biofuels from plant cell walls. However, proteomic characterization of the Golgi has so far been limited, owing to the technical challenges inherent in Golgi purification. In this study, a combination of density centrifugation and surface charge separation techniques have allowed the reproducible isolation of Golgi membranes from Arabidopsis (Arabidopsis thaliam) at sufficiently high purity levels for in-depth proteomic analysis. Quantitative proteomic analysis, immunoblotting, enzyme activity assays, and electron microscopy all confirm high purity levels. A composition analysis indicated that approximately 19% of proteins were likely derived from contaminating compartments and ribosomes. The localization of 13 newly assigned proteins to the Golgi using transient fluorescent markers further validated the proteome. A collection of 371 proteins consistently identified in all replicates has been proposed to represent the Golgi proteome, marking an appreciable advancement in numbers of Golgi-localized proteins. A significant proportion of proteins likely involved in matrix polysaccharide biosynthesis were identified. The potential within this proteome for advances in understanding Golgi processes has been demonstrated by the identification and functional characterization of the first plant Golgi-resident nucleoside diphosphatase, using a yeast complementation assay. Overall, these data show key proteins involved in primary cell wall synthesis and include a mixture of well-characterized and unknown proteins whose biological roles and importance as targets for future research can now be realized.

Abstract The present investigation was aimed to examine the percentage quantity of protein and amino acids in scales and muscles of Pampus argenteus and Parastromateus niger gathered from the local fish market of district Quetta of Balochistan. About 80 specimens of these two species, i.e., Pampus argenteus (N=40) and Parastromateus niger (N = 40), were collected from April 2017 to May 2018. In general, crude protein content was high in scales, that is, 71.03% in Parastromateus niger and 52.11% in Pampus argenteus, as well as in muscles of two Pomfret species of fishes i.e., 63.44% in Pampus argenteus and 60.99% in Parastromateus niger on a dry-weight basis, respectively. Likewise, the muscles and scales of Parastromateus niger reveal well compositions of amino acids that include proline was found to be high, and methionine was less than other amino acids, whereas threonine was found high in the scales of Pampus argenteus, but methionine was observed in lesser amount. However, the amino acids found in Pampus argenteus muscles also showed different compositions, such as lysine was found to be high, but histidine was less, respectively. In comparison, amino acids like tryptophan and cysteine were not detected in both scales and muscles of thesePomfret species of fishes. Thus, this study was based on analyzing the utilization of both Pomfret species of scales and meat whether they could have values as good supplements of both protein and certain kinds of essential amino acids in animal diets. Resumo A presente investigação teve como objetivo examinar a quantidade percentual de proteínas e aminoácidos em escamas e músculos de Pampus argenteus e Parastromateus niger coletados no mercado de peixes local do distrito de Quetta, no Baluchistão. Cerca de 80 exemplares dessas duas espécies, ou seja, Pampus argenteus (N=40) e Parastromateus niger (N=40), foram coletadas de abril de 2017 a maio de 2018. Em geral, o teor de proteína bruta foi alto em escamas, ou seja, 71,03% em Parastromateus niger e 52,11% em Pampus argenteus, bem como em músculos de duas espécies de peixes Pomfret, ou seja, 63,44% em Pampus argenteus e 60,99% em Parastromateus niger em base de peso seco, respectivamente. Da mesma forma, os músculos e escamas de Parastromateus niger revelam que as composições de aminoácidos que incluem prolina são altas, e a metionina é menor do que outros aminoácidos, enquanto a treonina foi encontrada alta nas escamas de Pampus argenteus, mas a metionina foi observada em quantidade menor. No entanto, os aminoácidos encontrados nos músculos de Pampus argenteus também apresentaram composições diferentes, como a lisina foi alta, mas a histidina foi menor, respectivamente. Em comparação, aminoácidos como triptofano e cisteína não foram detectados em ambas as escamas e músculos dessas espécies de peixes Pomfret. Assim, este estudo procurou analisar a utilização de ambas as espécies Pomfret de escamas e carne, e se eles poderiam ter valores como bons suplementos de proteína e certos tipos de aminoácidos essenciais em dietas de animais.

Abstract Salinity is one of the most critical environmental parameters regarding fish physiology, modifying food intake and growth performance in many fish species. The present study has investigated the effects of different salinity levels on growth performance, feeding and survival of Asian seabass Lates calcarifer juveniles. Asian seabass juveniles were reared at 0 (T1), 5 (T2), 22 (T3), 36 (T4), and 42 (T5) ppt salinity. Approximately eight hundred thirty fish individuals with an average weight of 1.24±0.52 g were randomly distributed (166 fish/Tank) in 5 concrete tanks (each tank 30×6×4 ft, volume 19,122 L) for forty days. Juveniles were initially fed 42% crude protein-containing diets at a rate of 6% of their body weight per day. The results showed that salinity level had a significant effect on the weight gain (WG), average daily weight gain (ADWG), specific growth rate (SGR), feed conversion ratio (FCR), survival rate (SR), total biomass and health indices (p<0.05). The highest WG (39.11±1.49 g), ADWG (1.00±0.12 g), SGR (8.74±0.03% d-1) and lowest FCR (0.96±0.20) were observed with T3 treatment, which was significantly higher compared to other treatment groups (p<0.05). Among the health indices, the highest hepatosomatic index and viscerosomatic index were found with T3 treatment, significantly higher than the other groups (p<0.05). No significant differences were found among the treatments in terms of survival rate (p>0.05), but the maximum survival rate (98.89±0.0%) was observed in the T3 and T2 treatments. The maximum level of crude proteins (19.99±1.4%) was found in the whole-body biochemical composition of Asian seabass juveniles in the T3 treatment group. The second-order polynomial regression showed that 20 ppt salinity is optimum for the best growth of Asian seabass. Thus, the present study recommends 20 to 36 ppt salinity for the commercial farming of Asian seabass under a closed aquaculture system. Resumo A salinidade é um dos parâmetros ambientais mais críticos em relação à fisiologia dos peixes, modificando a ingestão de alimentos e o desempenho de crescimento em muitas espécies. O presente estudo investigou os efeitos de diferentes níveis de salinidade no desempenho de crescimento, alimentação e sobrevivência de jovens espécies de robalos asiáticos Lates calcarifer, as quais foram criadas sob salinidade 0 (T1), 5 (T2), 22 (T3), 36 (T4) e 42 (T5) ppt. Aproximadamente 830 indivíduos de peixes com peso médio de 1,24 ± 0,52 g foram distribuídos aleatoriamente (166 peixes / tanque) em cinco tanques de concreto (cada tanque tinha 30 × 6 × 4 pés (9,1 x 1,8 x 1,2 metros), com volume de 19,12 litros) por 40 dias. Os peixes foram inicialmente alimentados com uma dieta contendo 42% de proteína bruta a uma taxa de 6% do seu peso corporal por dia. Os resultados mostraram que o nível de salinidade teve um efeito significativo no ganho de peso (GP), ganho de peso médio diário (GPMD), taxa de crescimento específico (TCE), taxa de conversão alimentar (TCA), taxa de sobrevivência (TS), biomassa total e índices de saúde (p < 0,05). O maior GP (39,11 ± 1,49 g), GPMD (1,00 ± 0,12 g), TCE (8,74 ± 0,03% d-1) e o menor TCA (0,96 ± 0,20) foram observados com o tratamento T3, que foi significativamente superior em comparação com os outros tratamentos (p < 0,05). Entre os índices de saúde, os maiores índices hepatossomáticos e viscerossomáticos foram encontrados no tratamento T3, significativamente superior do que os demais grupos (p < 0,05). Não foram encontradas diferenças significativas entre os tratamentos quanto à TS (p > 0,05), mas a TS máxima (98,89 ± 0,0%) foi observada nos tratamentos T3 e T2. O nível máximo de proteína bruta (19,99 ± 1,4%) foi encontrado na composição bioquímica dos corpos dos jovens robalos asiáticos no grupo T3. A regressão polinomial de segunda ordem mostrou que a salinidade de 20 ppt é ótima para o melhor crescimento do robalo asiático. Assim, o presente estudo recomenda salinidade de 20 a 36 ppt para o cultivo comercial de robalo asiático em sistema de aquicultura fechado.

Abstract Intestinal parasitism is the main cause of disease all over the world and described as a significant community health problem. The current study intended to find out the occurrence and identification of hazard factors linked with IPIs among 4-12 years aged shool-age children residing in Lower Dir district, Pakistan during 2019 - 2020. A cross-sectional school-based study was conducted using a pre-arranged pre-tested survey. Anthropometric data and stool collection were done to obtain the findings. The direct wet mount preparation in saline/iodine/methods was used for stool examination. Data were investigated using the GraphPad Prism 5. A total of 400 children studied (mean age of 8.6±3.6 years) the total incidence rate for the intestinal parasitic disease was established to be 71.75%. Of the 400 children studied, the overall prevalence rate for intestinal parasitic infections was found to 71.75% Ascaris lumbricoides (33.1%), Trichuris trichiura (1.04%), E. vermicularis (1.39%), Hookworm (19.86%) were identified in children living in the study area. We concluded that there is a mass scale campaigns were required to generate alertness about health and sanitation in children and the need for the development of effective poverty control programs because deworming (killing of worm with drugs) alone is not adequate to control parasitic infections. Resumo O parasitismo intestinal é a principal causa de doença em todo o mundo e descrito como um importante problema de saúde comunitária. O presente estudo buscou descobrir a ocorrência e identificação de fatores de risco relacionados a infecções parasitárias intestinais (IPIs) entre crianças em idade escolar de 4 a 12 anos residentes no distrito de Lower Dir, Paquistão durante 2019-2020. Um estudo transversal baseado em escola foi realizado usando uma pesquisa preestabelecida e pré-testada. Dados antropométricos e coleta de fezes foram feitos para obtenção dos achados. A preparação de montagem úmida direta em solução salina/iodo/métodos foi usada para exame de fezes. Os dados foram investigados usando o GraphPad Prism 5. Em um total de 400 crianças estudadas (idade média de 8,6±3,6 anos), a taxa de incidência total da parasitose intestinal foi estabelecida em 71,75%. Das 400 crianças estudadas, a taxa de prevalência global de infecções parasitárias intestinais foi de 71,75%. Ascaris lumbricoides (33,1%), Trichuris trichiura (1,04%), E. vermicularis (1,39%), Ancilostomíase (19,86%) foram identificados em crianças morando na área de estudo. Concluímos que há necessidade de campanhas em massa para gerar alerta sobre saúde e saneamento nas crianças, e a necessidade do desenvolvimento de programas efetivos de controle da pobreza, pois a desparasitação por si só não é adequada para controlar as infecções parasitárias.

Abstract Present study was conducted among school children to recognize the prevalence of IPIs in rural communities of district Dir Lower, Pakistan. A sum of 324 samples of stool were collected (210 boys and 114 girls). Used direct smear method and formol ether sedimentation concentration technique for processing the samples. The result shows that 82% (n=266) were found infected comprised 64.8% male and 35.1% females. Children of the age group 10 to 12 years were found extremely infected 94.2% while 4-6 year age group were having minimum ratio of infection 72%. Current study shows mono parasitism in 50.6% of the students while 22.2% were infected with 2 species and 7.40% were infected with three species of parasites. Seven species of intestinal parasites were reported include Ascaris lumbricoid in male (n=122) 58.0% and in female (n=65) 57.0% followed by Hook worm (n=88) 41.9% and (n=44) 38.5%; Tania saginata (n=44) 20.9% and (n=24) 21.0%; Entrobius vermicularis (n=32) 15.2% and (n=16) 14.0%; Trichuris trichura (n=25) 11.9% and (n=22) 19.2%; Hymenolepis nana (n=24) 11.4% and (n=18) 15.7% and Entameoba histolytica (n=16) 7.61% and (n=14) 12.2% in male and females respectively. The study indicates that most occurring intestinal parasite in the current study were Ascaris lumbricoides 58.0% (n=122) followed by hookworms 41.9% (n=88). Male students were more infected than females in the present study. Resumo O presente estudo foi conduzido entre crianças em idade escolar para reconhecer a prevalência de IPIs em comunidades rurais do distrito de Dir Lower, Paquistão. Foram coletadas 324 amostras de fezes (210 meninos e 114 meninas). Método de esfregaço direto usado e técnica de concentração de sedimentação de formol éter para processar as amostras. O resultado mostra que 82% (n = 266) dos infectados eram 64,8% do sexo masculino e 35,1% do feminino. Crianças da faixa etária de 10 a 12 anos foram encontradas extremamente infectadas 94,2%, enquanto a faixa etária de 4 a 6 anos apresentava proporção mínima de infecção de 72%. O estudo atual mostra monoparasitismo em 50,6% dos alunos, enquanto 22,2% estavam infectados com 2 espécies e 7,40% estavam infectados com três espécies de parasitas. Sete espécies de parasitas intestinais foram relatadas incluindo Ascaris lumbricoid em machos (n = 122) 58,0% e em fêmeas (n = 65) 57,0% seguido por anzol (n = 88) 41,9% e (n = 44) 38,5%; Tania saginata (n = 44) 20,9% e (n = 24) 21,0%; Entrobius vermicularis (n = 32) 15,2% e (n = 16) 14,0%; Trichuris trichura (n = 25) 11,9% e (n = 22) 19,2%; Hymenolepis nana (n = 24) 11,4% e (n = 18) 15,7% e Entameoba histolytica (n = 16) 7,61% e (n = 14) 12,2% em homens e mulheres, respectivamente. O estudo indica que os parasitas intestinais que mais ocorreram no presente estudo foram Ascaris lumbricoides 58,0% (n = 122), seguidos por ancilóstomos 41,9% (n = 88). Estudantes do sexo masculino foram mais infectados do que do sexo feminino no presente estudo.

Abstract The present study examines the correlations between fifteen morphometric and ten meristic characters and total length (TL) of males, females, and combined sexes of Alepes vari (Cuvier, 1833) collected from Karachi fish harbor, West Wharf of Karachi Coast. Statistical analyses of linear regression relationships show mostly strong correlations (r≥0.70; p<0.05) between total length (TL) and most morphometric characters in males, females, and combined sexes, except the height of pectoral-fin (PFH), and pelvic-fin base length (PelFL); whereas, meristic characters were found to be constant and indicate weak or negative type correlations (r≤0.50; p>0.05) with total length (TL). Hence, according to our present results, there is a direct relationship between the total length of fish and all morphometric characters, which were found to be the best indicators of positive allometric pattern growth in fish. Moreover, analysis of the 2-sample t-test revealed (t-test; p>0.05) that no sexual dimorphism was reported in Alepes vari. Thus, our present study could be valuable in systematic classification, sexual dimorphism, and management of this species on the Karachi coast. Resumo O presente estudo examina as correlações entre 15 caracteres morfométricos e 10 caracteres merísticos e comprimento total (CT) de machos, fêmeas e sexos combinados de Alepes vari (Cuvier, 1833), coletados do porto de Karachi, West Wharf, na costa de Karachi. As análises estatísticas das relações de regressão linear mostraram, principalmente, correlações fortes (r ≥ 0,70; p < 0,05) entre o CT e a maioria dos caracteres morfométricos em machos, fêmeas e sexos combinados, exceto a altura da nadadeira peitoral e o comprimento da base da nadadeira pélvica, enquanto os caracteres merísticos foram constantes, indicando correlações fracas ou negativas (r ≤ 0,50; p > 0,05) com o CT. Portanto, de acordo com nossos resultados, existe uma relação direta entre o CT dos peixes e todos os caracteres morfométricos, que foram considerados os melhores indicadores de crescimento do padrão alométrico positivo em peixes. Além disso, a análise do teste t de duas amostras revelou (teste t; p > 0,05) que nenhum dimorfismo sexual foi relatado em A. vari. Assim, o presente estudo pode ser valioso na classificação sistemática, dimorfismo sexual e manejo dessa espécie na costa de Karachi.

Abstract This study was conducted to estimate the diversity and the occurrence of commercially important finfish species collected by twenty fish sampling site of Sindh and Baluchistan coasts of the Arabian Sea in Pakistan from January to December 2019. Additionally, physicochemical characteristics of seawater were analyzed from these selected sites and found to be within suitable ranges required for fish growth and survive. A total of 81287 fish individuals were collected and identified as 49 species belonging to 26 families in our study. The most diversified family was Sparidae (13 species) followed by Carangidae and Lutjanidae (4 species), Mullidae, Serranidae, Ariidae (3 species), and Sciaenidae (2 species). The remaining 20 families were represented by only one species. The values of Shannon diversity index calculated for the four selected habitats revealed that high fish diversity was reported at Sonmiani Coast (H'=1.81), while less at Ormara Coast (H'=0.23). Likewise, Evenness index (E) was high at Sonmiani Coast (E=0.50) and less fish diversity was reported at Ormara Coast (E=0.06). Reducing risks to threatened marine species in coastal habitats also requires conservation actions at multiple scales. Thus, it was concluded that our study could be valuable in providing the more information’s regarding to the diversity of finfish species and their occurrence along the Pakistan Coast. Further, to better understand the effects, regular monitoring and conservation measures should be taken to mitigate the influence of anthropogenic activities and protect finfish diversity from further decline Resumo Este estudo foi conduzido para estimar a diversidade e a ocorrência de espécies de peixes comercialmente importantes coletadas por vinte locais de amostragem de peixes nas costas de Sindh e Baluchistão do mar da Arábia, no Paquistão, de janeiro a dezembro de 2019. Além disso, as características físico-químicas da água do mar foram analisadas a partir desses peixes locais selecionados e considerados dentro dos intervalos adequados necessários para o crescimento e sobrevivência dos peixes. Um total de 8.1287 indivíduos de peixes foi coletado e identificado como 49 espécies pertencentes a 26 famílias em nosso estudo. A família mais diversificada foi Sparidae (13 espécies), seguida por Carangidae e Lutjanidae (4 espécies), Mullidae, Serranidae, Ariidae (3 espécies) e Sciaenidae (2 espécies). As 20 famílias restantes foram representadas por apenas uma espécie. Os valores do índice de diversidade de Shannon calculados para os quatro habitats selecionados revelaram que uma alta diversidade de peixes foi relatada na costa Sonmiani (H' = 1,81), enquanto menos na costa Ormara (H' = 0,23). Da mesma forma, o índice de regularidade (E) foi alto na costa de Sonmiani (E = 0,50) e menos diversidade de peixes foi relatada na costa de Ormara (E = 0,06). A redução dos riscos para as espécies marinhas ameaçadas em habitats costeiros também requer ações de conservação em várias escalas. Assim, concluiu-se que nosso estudo pode ser valioso para fornecer mais informações sobre a diversidade de espécies de peixes finos e sua ocorrência ao longo da costa do Paquistão. Além disso, para compreender melhor os efeitos, medidas regulares de monitoramento e conservação devem ser tomadas para mitigar a influência das atividades antropogênicas e proteger a diversidade de peixes finos de um declínio maior.