Abstract Heart and blood vessel disorders, such as coronary heart disease, brain vessel disease, rheumatic heart disease, and others, are together referred to as cardiovascular disease (CVD). In this study, we sought to determine how mitochondrial Leucine Transfer RNA genes and CVDs are related (MT-L1 and MT-L2). From CVD patients in Peshawar, a total of 27 saliva samples were taken. Leu-tRNA genes expressed by mitochondria were amplified using polymerase chain reaction after DNA was removed. Ten samples were sent for sequencing after PCR and gene cleaning. We obtained all of the sequenced results, which were subsequently aligned and evaluated against the mitochondrial revised Cambridge Reference Sequence (rCRS). However, in our sequenced samples, Leu-tRNA MT-L1 and MT-L2 genes were determined to be unaltered. Thus, it is suggested that a large population be taken into account while screening for mutations in the mitochondrial encoded Leu-tRNA MT-L1 and MT-L2 genes of cardiac patients in areas of Pakistan. Additionally, it is recommended that patients with cardiac problems should also have other mitochondrial encoded genes checked for potential mutations. This could result in the identification of genetic markers that could be used for early CVD screening in Pakistan. disorders others CVD. . (CVD) study MTL1 MTL MT L1 L (MT-L MTL2. MTL2 L2 MT-L2) Peshawar 2 LeutRNA Leu tRNA removed cleaning results rCRS. rCRS (rCRS) However MT-L unaltered Thus Pakistan Additionally (CVD (rCRS

Resumo Distúrbios do coração e dos vasos sanguíneos, como doença cardíaca coronária, doença dos vasos cerebrais, doença cardíaca reumática entre outros, são referidos juntos como doença cardiovascular (DCV). Neste estudo, procuramos determinar como os genes mitocondriais do RNA de transferência de leucina e as DCVs estão relacionados (MT-L1 e MT-L2). Foi coletado um total de 27 amostras de saliva de pacientes com DCV em Peshawar. Genes de Leu-tRNA expressos por mitocôndrias foram amplificados usando reação em cadeia da polimerase (PCR) após a remoção do DNA. Dez amostras foram enviadas para sequenciamento após PCR e limpeza gênica. Obtivemos todos os resultados sequenciados, que foram posteriormente alinhados e avaliados em comparação com a Sequência de Referência de Cambridge revisada (rCRS). No entanto, em nossas amostras sequenciadas, os genes Leu-tRNA MT-L1 e MT-L2 foram determinados como inalterados. Assim, sugere-se que uma grande população seja levada em consideração durante a triagem de mutações nos genes Leu-tRNA MT-L1 e MT-L2 mitocondriais codificados de pacientes cardíacos em áreas do Paquistão. Além disso, recomenda-se que outros genes mitocondriais codificados de pacientes com problemas cardíacos também sejam verificados quanto a possíveis mutações. Isso pode resultar na identificação de marcadores genéticos que podem ser usados para triagem precoce de DCV no Paquistão. sanguíneos coronária cerebrais DCV. . (DCV) estudo MTL1 MTL MT L1 L (MT-L MTL2. MTL2 L2 MT-L2) 2 Peshawar LeutRNA Leu tRNA (PCR DNA gênica sequenciados rCRS. rCRS (rCRS) entanto sequenciadas MT-L inalterados Assim sugerese sugere se Paquistão disso recomendase recomenda (DCV (rCRS

Resumo O peixe é a principal fonte de proteína animal para a alimentação humana. O objetivo deste estudo foi descobrir a prevalência de bactérias patogênicas de dois peixes economicamente importantes selecionados do Paquistão, nomeadamente Mahseer (Tor putitora) e carpa prateada (Hypophthalmichthys molitrix). Amostras de peixes vivos de incubatórios e amostras de peixes mortos de diferentes mercados da área de estudo foram coletadas aleatoriamente. As amostras de peixes foram analisadas quanto ao isolamento, identificação e prevalência de bactérias. As bactérias isoladas dos peixes do estudo foram identificadas através de testes bioquímicos e cerca de 10 espécies de bactérias patogênicas foram identificadas incluindo as bactérias patogênicas para humanos e peixes, nomeadamente, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus iniae, Serratia spp. Citrobacter spp. Stenotrophomonas spp. Bacillus spp. e Salmonella spp. A porcentagem de freqüência de ocorrência bacteriana em carpa prateada e peixes Mahseer mostrou Pseudomonas aeruginosa 21,42%, Staphylococcus epidermidis 17,85%, Escherichia coli 11,90%, Staphylococcus aureus 9,52%, Citrobacter spp. 9,52%, Serratia spp. 8,33%, Streptococcus iniae 7,14%, Stenotrophomonas spp. 5,95%, Bacillus spp. 4,76% e Salmonella spp. 3,57%. O estudo revelou que as amostras de peixes de Mahseer e carpa prateada coletadas nos mercados encontraram mais isolados (10 espécies bacterianas) do que as amostras de peixes frescos (03 espécies bacterianas) de incubatórios. A ocorrência de bactérias patogênicas nos peixes do estudo apresentou fator de risco para consumidores de saúde pública.

Abstract Fish is the main source of animal protein for human diet. The aim of this study was to find out prevalence of pathogenic bacteria of two selected economically important fish of Pakistan namely Mahseer (Tor putitora) and Silver carp (Hypophthalmichthys molitrix). Live fish samples from hatcheries and dead fish samples from different markets of study area were randomly collected. The fish samples were analyzed for isolation, identification and prevalence of bacteria. The isolated bacteria from study fish were identified through biochemical test and about 10 species of pathogenic bacteria were identified including the pathogenic bacteria to human and fish namely, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus iniae, Serratia spp. Citrobacter spp. Stenotrophomonas spp. Bacillus spp. and Salmonella spp. The bacterial percentage frequency of occurrence in Silver carp and Mahseer fish showed Pseudomonas aeruginosa 21.42%, Staphylococcus epidermidis 17.85%, Escherichia coli 11.90%, Staphylococcus aureus 9.52%, Citrobacter spp. 9.52%, Serratia spp. 8.33%, Streptococcus iniae 7.14%, Stenotrophomonas spp. 5.95%, Bacillus spp. 4.76% and Salmonella spp. 3.57%. The study revealed that Fish samples of Mahseer and Silver carp that were collected from markets have found more isolates (10 bacterial species) than did the fresh fish pond samples (03 bacterial species) of hatcheries. The occurrence of pathogenic bacteria in study fish showed risk factor for public health consumers.

Resumo Juglans regia L. são plantas frutíferas nutritivas encontradas nas áreas do norte do Paquistão. A população de nozes foi explorada do distrito de Dir e Swat, Khyber Pakhtunkhwa, Paquistão por sua divergência geográfica, climática e química. As diferenças geográficas, como altitude, latitude e longitude, enquanto as diferenças climáticas viz. Ec do solo, pH do solo, precipitações, intensidade de luz, temperatura e temperatura do solo. Em ambos os distritos, o TPC variou de 211,2±0,6 a 227,8±0,4 mg/100g, o RSA variou de 43,32±1,5% a 52,18±0,4%, a condutividade variou de 296,43±0,6 a 312,22±0,3 S/m e composição elementar como ferro, cobre, cálcio, zinco e magnésio em Dir difere de 0,312±0,032, 0,209±0,13, 20,0±0,313, 0,406±0,10 e 10,2±0,030 mg/L a 0,543±0,65, 0,698±0,82, 28,7±0,234, 0,685±0,15 e 17,6±0,015 mg/L respectivamente. A altitude e a temperatura mostraram correlação com os teores de fenólicos totais e atividade de eliminação de radicais, enquanto o pH do solo, precipitações, temperatura do solo, Ec do solo e intensidade de luz indicaram uma correlação fraca com características químicas das nozes. Mais estudos de nozes são necessários para explorar seus fitoquímicos terapeuticamente importantes para obter naturalmente nutrientes farmacêuticos de máxima importância para a saúde dos seres humanos.

Abstract Juglans regia L. are nutritious fruit bearing plants mostly found in Northern areas of Pakistan. The population of walnuts was explored from district Dir and Swat Khyber Pakhtunkhwa, Pakistan for their geographical, climatic and chemical divergence. The geographical differences such as altitude, latitude and longitude whereas climatic differences viz. soil EC, soil pH, precipitations, intensity of light, temperature and soil temperature. In both districts TPC ranged from 211.2±0.6 to 227.8±0.4 mg/100g, RSA ranged from 43.32±1.5% to 52.18±0.4%, conductivity ranged from 296.43±0.6 to 312.22±0.3 S/m and elemental composition such as iron, copper, calcium, zinc and magnesium in Dir differs from 0.312±0.032, 0.209±0.13, 20.0±0.313, 0.406±0.10 and 10.2±0.030 mg/L to 0.543±0.65, 0.698±0.82, 28.7±0.234, 0.685±0.15 and 17.6±0.015 mg/L respectively. Altitude and temperature showed a correlation with total phenolics contents and radical scavenging activity while soil pH, precipitations, soil temperature, soil Ec and light intensity indicated a weak correlation with chemical traits of walnuts. Further studies of walnuts are needed to explore their therapeutically important phytochemicals to succeed naturally pharmaceutical nutrients of the maximum significance for the health of human beings.

This study sought to explore the effect of the administration of long-term, exogenous oxytocin on buffalo milk. In this study, newly-calved postpartum buffaloes (Nili-Ravi) (n = 24) were divided equally into three groups: (control without oxytocin - CON, 10 i.u. oxytocin - LOW, 30 i.u. oxytocin - HIGH). Oxytocin was injected twice a day before each 154-day milking. Milk production, milk composition, somatic cell counts, and progesterone (P4) were evaluated daily, weekly, and fortnightly, respectively. Results revealed that the HIGH group produced high milk production and pH values, but lower milk density than the CON and LOW groups. The HIGH group and LOW group had higher milk fat and solid-not-fat (SNF) compared to the CON group. SNF and protein were higher in the LOW group than the CON group. Freezing point, lactose, solids, somatic cell count, and P4 were similar in all three groups. Overall, it can be concluded that long-term, exogenous oxytocin has a positive effect on milk production, fat, SNF, and protein, with a negative effect on milk pH. None of the oxytocin treatments affected somatic cell counts and P4 concentration in postpartum buffalo milk.

Resumo O vírus da hepatite C (HCV) é o grave problema de saúde pública das doenças hepáticas. Aproximadamente 170 milhões de pessoas no mundo estão infectadas com HCV; no Paquistão, a doença tem alto índice de ocorrência. O presente estudo prevê uma prevalência atualizada do HCV e distribuição genotípica na população geral do distrito de Mardan, Khyber Pakhtunkhwa (KP), Paquistão. As amostras de sangue de 6.538 indivíduos, incluindo 3.263 homens e 3.275 mulheres, foram analisadas para o antígeno de superfície da hepatite C por teste imunocromatográfico (ICT), ensaio imunoenzimático (ELISA) e reação em cadeia da polimerase de transcrição reversa (PCR). Verificou-se que 396 (12,13%) de 3.263 indivíduos continham anticorpos no sangue contra o HCV, enquanto entre as diferentes faixas etárias as maiores incidências de anticorpos anti-HCV foram encontradas na faixa etária de 31 a 40 anos (11,01%). As amostras positivas para ICT foram posteriormente rastreadas por nested PCR para determinar a existência de HCV-RNA ativo. Identificou-se que 7,11% (3.263) do total da população (6.538) testada foram positivos, dentre os quais 461 (14,07%) mulheres possuíam anticorpos no sangue contra o HCV. Nossos dados mostraram que a infecção total pelo HCV na população investigada foi de 5,78%. Maior porcentagem de prevalência de HCV foi detectada em homens do que em mulheres nas faixas etárias de 31-40 e 41-50. Para comparar a prevalência de genótipos de HCV com relação à idade no genótipo masculino e feminino 3a foi encontrado o genótipo mais prevalente seguido por 1a, 2a e 3b, respectivamente.

Abstract Hepatitis C virus (HCV) is the serious global public health burden of liver disease. Approximately 170 million people in the world are infected with (HCV). In Pakistan, where the disease has high occurrence rate. The present study envisages an up-to-date prevalence of HCV and genotypic distribution in the general population of Mardan District, Khyber Pakhtunkhwa (KP), Pakistan. The blood samples from 6,538 individuals including 3,263 males and 3,275 females were analyzed for hepatitis C surface antigen by Immuno-chromatographic test (ICT), Enzyme-linked immunosorbent assay (ELISA), and reverse transcription-polymerase chain reaction (PCR). It was found that 396 (12.13%) out of 3263 individuals contained antibodies in their blood against HCV, while among the different age groups, the highest incidences of HCV antibodies were found in the 31-40 age group (11.01%). The ICT positive samples were further screened by nested PCR to determine the existence of active HCV-RNA. It was identified that 7.11% (3263) of the total population (6538) tested was positive, among which the 461 (14.07%) females possessed antibodies in their blood against HCV. Our data showed total HCV infection in the investigated population was 5.78%. Higher percentage of HCV prevalence was detected in males than females in the age group 31-40 and 41-50. To compare the prevalence of HCV genotypes age-wise in male and female genotype 3a was found most prevalent genotype followed by 1a, 2a and 3b, respectively.

Resumo Os manipuladores de alimentos desempenham um papel fundamental na transmissão de protozoários e helmintos parasitas patogenicamente importantes. Este estudo teve como objetivo avaliar a prevalência de protozoários patogênicos intestinais e helmintos parasitas entre manipuladores de alimentos na Universidade de Malakand, Lower Dir, Paquistão. Amostras de fezes foram coletadas de 642 manipuladores de alimentos (todos do sexo masculino) em um estudo transversal de janeiro a novembro de 2017. Técnicas de montagem úmida e métodos de concentração usando soluções de sal e formol-éter. Trezentos e oitenta e quatro casos (59,8%) foram encontrados infectados com mais um parasita. A maioria dos indivíduos foi encontrada infectada por helmintos (47,6%) em comparação com protozoários intestinais (0,93%). Setenta e dois casos (11,2%) dos casos apresentavam infecção mista com protozoários intestinais e helmintos parasitas. A ordem de prevalência de helmintos intestinais foi Ancylostoma duodenale (n = 258, 40,1%), seguido por Taeniasa ginata (n = 96, 14,9%) Ascaris lumbricoides (n = 54, 8,40%) e Trichuris trichura (n = 30, 4,60 %). Para protozoários intestinais, Entamoeba histolytica / dispar (n = 36, 5,64%) foi o único protozoário detectado. Monoparasitismo foi maior do que poliparasitismo. A renda familiar e o nível de escolaridade foram os fatores significativamente (P <0,05) associados na prevalência de parasitos. A pesquisa atual mostrou que os IPIs são principalmente os patógenos de origem alimentar, ainda um importante problema de saúde pública no Paquistão. Programas eficazes de controle da transferência de doenças parasitárias e seus fatores associados são recomendados.

Abstract Food handlers plays a primary role in the transmission of pathogenically important protozoans and helminth parasites. This study was aimed to evaluate the prevalence of intestinal pathogenic protozoans and helminth parasites among food handlers in and around University of Malakand, Lower Dir, Pakistan. Stool samples were collected from 642 food handlers (all of male) in a cross-sectional study from January to November, 2017. Wet Mount Techniques and concentration methods by using salt and formol–ether solutions. Three hundred and eighty four cases (59.8%) were found infected with one more parasites. Most of the individuals were found infected with helminth (47.6%) as compared to intestinal protozoans (0.93%). Seventy two cases (11.2%) of the cases presented mixed infection with both intestinal protozoan and helminth parasites. The order of prevalence for intestinal helminth was Ancylostoma duodenale (n = 258, 40.1%), followed by Taeniasa ginata (n=96, 14.9%) Ascaris lumbricoides (n = 54, 8.40%) and Trichuris trichura (n=30, 4.60%). For intestinal protozoa, Entamoeba histolytica/dispar (n = 36, 5.64%) was the only protozoan detected. Mono-parasitism was higher than poly-parasitism. Family size income and education level were the factors significantly (P<0.05) associated in the parasites prevalence. Current research showed that IPIs are primarily the foodborne pathogens still an important public health problem in Pakistan. Effective control programs on parasitic diseases transfer and their associated factors are recommended.

Resumo A infecção pelo vírus da hepatite B é perigosa entre os cinco tipos de hepatite, pois permanece clinicamente assintomática. O presente estudo traça a prevalência atualizada do vírus da hepatite B (HBV) na população geral de Mardan, Khyber Pakhtunkhwa, no Paquistão. Amostras de sangue de 4.803 indivíduos, incluindo 2.399 homens e 2.404 mulheres, foram investigadas. Todas as amostras suspeitas foram analisadas para o antígeno de superfície da hepatite B usando teste imunocromatográfico (ICT), enzyme-linked immunosorbent assay (ELISA), seguido por transcrição reversa-reação em cadeia da polimerase (RT-PCR). Os resultados mostraram que 312 (13,00%) de 2.399 indivíduos continham anticorpos no sangue contra o VHB, enquanto, entre as diferentes faixas etárias, as maiores incidências de anticorpos VHB foram encontradas nos grupos de 21 a 30 anos (10,73%). Além disso, amostras positivas para ICT foram rastreadas por reação em cadeia da polimerase aninhada para detectar a existência de HBV-DNA ativo. Observou-se que 169 (7,04%) de 2.399 homens do total da população (4803) testados foram positivos. Por outro lado, 463 mulheres (19,25%) possuíam anticorpos no sangue contra VHB. Acumulativamente, nossos resultados mostraram uma porcentagem maior de prevalência de HBV em homens do que em mulheres na faixa etária de 21 a 30 anos. O total de HCV infectados na população geral de Mardan foi registrado em 5,7%, incluindo homens e mulheres.

Abstract Hepatitis B virus infection is perilous among the five types of Hepatitis, as it remains clinically asymptomatic. The present study draws up-to-date prevalence of Hepatitis B virus (HBV) in the general population of Mardan, Khyber Pakhtunkhwa Pakistan. The blood samples from 4803 individuals including 2399 male and 2404 females were investigated. All the suspected samples were analyzed for hepatitis B surface antigen using Immuno-chromatographic test (ICT), Enzyme-linked immunosorbent assay (ELISA), and followed by Reverse transcription-polymerase chain reaction (RT-PCR). Results showed that 312 (13.00%) out of 2399 individuals contained antibodies in their blood against HBV, while among the different age groups, the highest incidences of HBV antibodies were found in the age of 21-30 groups (10.73%). Furthermore, the ICT positive samples were screened by nested polymerase chain reaction to detect the existence of active HBV-DNA. It was observed that 169 (7.04%) out of (2399) male of the total population (4803) tested was positive. On the other hand, the female 463 (19.25%) possessed antibodies in their blood against HBV. Accumulatively, our results showed a higher percentage of HBV prevalence in males than females in the age group 21-30 years. The total HCV infected in Mardan general population was recorded at 5.7% comprising both male and female.

Resumo A região de Malakand é uma área endêmica para leishmaniose cutânea (CL). No entanto, há um número limitado de estudos sobre esta doença no Paquistão. Portanto, foi realizado um estudo para entender o nível de atitude e prática de conscientização entre os residentes de Makaland em relação aos CL e os vetores da doença. Este estudo adotou uma abordagem transversal com um total de 400 entrevistados (n = 93 rural e n = 307 urbano). No geral, a população da região de Malakand (61,2%) estava bem informada sobre o papel da mosca na transmissão de doenças, mas a maioria não possui conhecimento sobre o comportamento do vetor e quase um quarto (24,5%) foi incapaz de fornecer conhecimento sobre medidas de controle adequadas . De maneira alarmante, a prática e as atitudes da população em geral não foram satisfatórias, pois cerca da metade (49,8%) da população adotou algum método de controle. Este estudo apela ao aumento da conscientização por meio de campanhas de educação em saúde para reduzir o risco de surtos de leishmaniose cutânea no futuro.

Abstract Malakand region is an endemic area for cutaneous leishmaniasis (CL). However, there are limited number of studies of this disease in Pakistan. Therefore, a study was conducted to understand the level of awareness attitude and practice among the residents of Makaland towards CL and the disease vectors. This study adopted a cross-sectional approach with a total of 400 respondents (n=93 rural and n= 307 urban). Overall, the population in Malakand region (61.2%) were well-informed in the role of sand fly in transmitting diseases, but most lack knowledge on the vector’s behavior and almost a quarter (24.5%) were unable to provide knowledge on proper control measures. Alarmingly, the practice and attitudes of the general population was not satisfactory as close to half (49.8%) of the population did adopt any control method. This study calls for increase in awareness through health education campaign to reduce the risk of cutaneous leishmaniasis outbreaks in the future.

Resumo A síntese verde foi introduzida como uma alternativa à síntese química devido às graves consequências. As nanopartículas metálicas sintetizadas através da abordagem verde têm diferentes aplicações farmacêuticas, médicas e agrícolas. O presente estudo seguiu uma rota verde e simples para a preparação de nanopartículas de ouro potencialmente bioativas (Au NPs). As NPs de Au foram preparadas via abordagem de síntese verde usando a porção alcaloide básica bruta do tubérculo de Delphinium chitralense. As NPs de Au sintetizadas verdes foram caracterizadas por microscopia eletrônica de transmissão (TEM), microscopia eletrônica de varredura (MEV), difração de raios X (DRX), infravermelho com transformada de Fourier (FTIR) e espectrofotômetro UV-Visível. A análise morfológica mostra que as NPs de Au possuem geometria cúbica com tamanhos diferentes. A análise espectroscópica UV-Vis confirmou a síntese de Au NPs enquanto a XRD provou sua fase cristalina pura. O Au NPs mostrou inibição dependente da dose promissora de AChE e BChE em comparação com a droga bruta e padrão.

Abstract Green synthesis has been introduced as an alternative to chemical synthesis due to the serious consequences. Metal nanoparticles synthesized through green approach have different pharmaceutical, medical and agricultural applications. The present study followed a green and simple route for the preparation of potentially bioactive gold nanoparticles (Au NPs). Au NPs were prepared via green synthesis approach using crude basic alkaloidal portion of the tuber of Delphinium chitralense. The green synthesized Au NPs were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD) fourier transform infrared (FTIR), and UV-Visible spectrophotometer. Morphological analysis shows that Au NPs have cubic geometry with different sizes. UV-Vis spectroscopic analysis confirmed the synthesis of Au NPs while XRD proved their pure crystalline phase. The Au NPs showed promising dose dependent inhibition of both AChE and BChE as compared to the crude as well as standard drug.

ABSTRACT Objective: Despite the presence of multitude of synthetic drugs against fever and inflammation, none has been proven entirely safe. In contrast, the accepted safety of plant derived natural products is inspiring the world. Based on this fact as well as in view of the diversified activities reported from the genus Gymnosporia, the present study was designed to evaluate the antipyretic and anti-inflammatory activity of Gymnosporia royleana (G royleana). Methods: The methanolic extract of the aerial parts of G royleana was screened for in-vivo antipyretic activity using the brewer's yeast-induced pyrexia mice model and for anti-inflammatory activity using the carrageenan-induced paw oedema and xylene-induced ear oedema mice model. Results: In the antipyretic assay, G royleana extract showed considerable antipyretic activity in a dose dependent fashion. Statistically significant antipyretic effects (p < 0.05) were observed at the end of the second hour of administration for all doses of extract and remained significant until the end of the experiment. The plant extract also displayed promising anti-inflammatory activity, in a dose dependent fashion, in both models of inflammation ie carrageenan- and xylene-induced oedema models, when compared to the controls. In the carrageenan-induced oedema model, significant effects (p < 0.01) were observed for 300 and 600 mg/kg doses after 60 minutes of xylene administration (ie 55.51% and 65.88% inhibition of oedema, respectively). Conclusion: The study provided evidence supporting the antipyretic and anti-inflammatory activity of the G royleana methanolic extract.

RESUMEN Objetivo: A pesar de la presencia de multitud de fármacos sintéticos en el arsenal contra la fiebre y la inflamación, ninguno ha dado pruebas de ser completamente seguro. En contraste con ello, la seguridad aceptada de los productos naturales derivados de las plantas inspira al mundo. Sobre la base de este hecho, así como en vista de las actividades diversificadas que se reportan con respecto al género Gymnosporia, el presente estudio se diseñó con el objeto de evaluar el potencial antipirético y antiinflamatorio de Gymnosporia royleana (G royleana). Métodos: El extracto de metanol de las partes aéreas de G royleana fue tamizado en busca de actividad antipirética in vivo, utilizando el modelo de pirexia inducida por levadura de cerveza en ratones, y de actividad antiinflamatoria utilizando modelos de ratones con oedema de las patas inducido mediante carragenina, y oedema de las orejas inducido mediante xileno. Resultados: En el ensayo antipirético, el extracto de G royleana mostró una actividad antipirética considerable en forma dependiente de la dosis. Se observó un efecto antipirético estadísticamente significativo (p < 0.05) en el transcurso de la segunda hora de administración para todas las dosis de extracto y se mantuvo significativo hasta el final del experimento. El extracto de la planta también mostró una actividad antiinflamatoria prometedora, de una manera dependiente de la dosis, en ambos modelos de inflamación, es decir, modelos de oedema inducido por carragenina y xileno, en comparación con el control. En el modelo de oedema inducido por carragenina, se observó un efecto significativo (p < 0.01) para dosis de 300 y 600 mg / kg después de 60 minutos de administración de xileno (es decir, 55.51% y 65.88% de inhibición del oedema, respectivamente). Conclusión: El estudio proporcionó pruebas suficientes sobre el potencial antipirético y antiinflamatorio del extracto de G royleana.

ABSTRACT Objective: In the present study, five new source compounds isolated from aerial parts of Gymnosporia royleana (G royleana) were screened for antibacterial and antifungal activities. Methods: Extraction from plant material was carried out using cold maceration technique. Isolation of pure compounds was accomplished through repeated column chromatography of different fractions obtained from crude extract and using silica gel as stationary phase. Their structures were established via advanced spectroscopic techniques along with the spectral data previously reported for these compounds. Dilution method was used for the evaluation of antimicrobial potential of the compounds against various microbial strains. Results: Among the tested compounds, Gymnosporin B displayed moderate antimicrobial activity against Escherichia Coli (E coli), Staphylococcus aureus (S aureus), Candida albicans (C albicans) and Aspergillus flavus (A flavus) [minimum inhibitory concentration (MIC) range; 32–64 μg/mL]. Similarly, Gymnosporin C also showed moderate activity against E coli and S aureus (MIC; 32 μg/mL each) as well as weak activity against C albicans and A flavus (MIC; 64 μg/mL each). However, Royaflavone showed moderate antibacterial activity against S aureus only (MIC; 32 μg/mL). Antimicrobial activity of the rest of the compounds was weak and negligible. Conclusion: The present study has provided fascinating results of antimicrobial activities of the isolated compounds. However, the broad antimicrobial spectrum of Gymnosporin B and Gymnosporin C demands for further exploration of these triterpenes, both on the basis of mechanism and quantitative structure-activity relationship.

RESUMEN Objetivo: En el presente estudio, cinco nuevos compuestos de origen aislados de partes aéreas de Gymnosporia royleana (G royleana) fueron tamizados en sus actividades antibacterianas y antifúngicas. Métodos: La extracción de material vegetal se realizó mediante la técnica de maceración en frío. El aislamiento de compuestos puros se logró a través de la cromatografía en columna repetida de diversas fracciones obtenidas del extracto crudo y usando gel de silicona como fase estacionaria. Sus estructuras fueron establecidas mediante técnicas espectroscópicas avanzadas junto con los datos espectrales previamente reportados para estos compuestos. El método de dilución fue usado para evaluar el potencial antimicrobiano de los compuestos contra diversas cepas microbianas. Resultados: Entre los compuestos sometidos a prueba, Gymnosporina B mostró una actividad antimicrobiana moderada contra Escherichia Coli (E coli), Staphylococcus aureus (S aureus), Candida Albicans (C albicans) y Aspergillus flavus (A flavus) [rango de concentración inhibitoria mínima (CIM); 32 – 64 μg/mL]. De manera similar, Gymnosporina C también mostró actividad moderada contra E coli y S aureus (CIM; 32 μg/mL cada uno) así como débil actividad frente a C albicans y A flavus (CIM; 64 μg/mL cada uno). Sin embargo, Royaflavone mostró actividad antibacteriana moderada sólo frente a S aureus (CIM; 32 μg/mL). La actividad antimicrobiana del resto de los compuestos fue débil e insignificante. Conclusión: El presente estudio ha proporcionado resultados interesantes acerca de las actividades antimicrobianas de los compuestos aislados. Sin embargo, el amplio espectro antimicrobiano de la Gymnosporina B y la Gymnosporina C exige una mayor exploración de estos triterpenos, tanto sobre la base del mecanismo como a partir de la relación cuantitativa estructura-actividad.

ABSTRACT Objective: Gymnosporia royleana (G royleana) Wall ex MA Lawson, locally known as (Sur Azghee), is traditionally used for the management of various diseases. In the current investigation, we made an effort to scientifically validate its traditional use in various pathological conditions, such as microbial infections and cancer, and to explore its additional pharmacological activities via random screening against locally accessible pharmacological methods, irrespective of its traditional uses like antidiabetic, haemagglutination and antioxidant assays. Methods: Extraction was carried out using a cold maceration methodology. Dilution method was used for antimicrobial susceptibility testing using different concentrations. Streptozocin (STZ) induced protocol was used to assess antidiabetic activity at a dose level of 200, 400 mg/ kg. Antioxidant activity, haemagglutination activity, and anticancer activities against HepG-2 and MCF-7 cell lines were determined as per established protocols. Similarly, the maximum amount of phenolic content (12.02 mg 100 g) was determined by using Folin Ciocalteu assay. Results: Promising antimicrobial activities in terms of minimum inhibitory concentration (MIC) were noted for crude extract (25–200 µg/mL), n-hexane (100–400 µg/mL), ethyl acetate (50–200 µg/mL) and aqueous (100–400 µg/mL). Antidiabetic potential was significant at a dose level of 200–400 mg/kg bodyweight by reducing the blood glucose level at days 10 and 15. The percentage of 2,2-diphenyl-1-picrylhydrazyl (DPPH) values increase by increasing the concentration of the plant extract (10–100 µg/mL). The methanol extract was found to possess high agglutination activity. Conclusion: It was concluded that this plant species possess significant antimicrobial, antidiabetic, antioxidant, anticancer and haemagglutination activities, which could be attributed to the phenolic content of the extract.

RESUMEN Objetivo: Gymnosporia royleana (G royleana) Wall ex MA Lawson, localmente conocida como “Sur Azghee”, se utiliza tradicionalmente para el tratamiento de diversas enfermedades. En la investigación presente, tratamos de validar científicamente su uso tradicional en varias condiciones, tales como las infecciones microbianas y el cáncer, así como explorar sus actividades farmacológicas adicionales mediante el tamizado aleatorio frente a los métodos farmacológicos localmente accesibles, independientemente de sus usos tradicionales como ensayos antidiabéticos, hemaglutinantes, y antioxidantes. Métodos: La extracción se realizó mediante una metodología de maceración en frío. Un método de dilución se utilizó para la prueba de susceptibilidad antimicrobiana utilizando diferentes concentraciones. Se utilizó el protocolo inducido por estreptozotocina (STZ) para evaluar la actividad antidiabética a un nivel de dosis de 200, 400 mg/kg. La actividad antioxidante, la actividad de hemaglutinación, y las actividades anticancerígenas contra las líneas celulares HepG-2 y MCF-7, se determinaron según los protocolos establecidos. De modo similar. la cantidad máxima de contenido fenólico (12.02 mg 100 g) se determinó mediante el uso del ensayo Folin-Ciocalteu. Resultados: Se observaron actividades antimicrobianas prometedoras en términos de la concentración inhibitoria mínima (CIM) para el extracto crudo (25–200 μg/mL), el n-hexano (100–400 μg/mL), el acetato de etilo (50–200 μg/mL), y el extracto acuoso (100–400 μg/mL). El potencial antidiabético fue significativo a un nivel de dosis de 200–400 mg/kg de peso corporal mediante la reducción del nivel de glucosa en sangre a los 10 y 15 días. El porcentaje de los valores de 2,2-difenil-1-picrilhidracilo (DPPH) se incrementa al aumentar la concentración del extracto de la planta (10–100 μg/mL). Se halló que el extracto de metanol posee una alta actividad de aglutinación. Conclusión: Se concluyó que esta especie de planta posee importantes actividades antimicrobianas, antidiabéticas, antioxidantes, anticancerígenas y hemaglutinantes, que podrían atribuirse al contenido fenólico del extracto.

ABSTRACT Although in-feed antibiotics work for broiler chicken´s growth, they are a source of public health hazard. Therefore, there is a need for alternates which can act as growth promoter without deleterious effects on the health of meat consumers. Moringa oleifera is one such phytobiotic which is reported to possess antimicrobial and immuno-modulatory properties. This study investigated the effect of Moringa oleifera leaf powder (MOLP) supplementation on meat quality and bone morphometry of broiler. One-day-old chicks (n=100), divided into five groups (four replicates with n=5), were fed a basal diet (control group) or same diet supplemented with 6, 9, 12 or 15 g/kg MOLP. On d-35, two birds per replicate were euthanized to collect samples of breast muscle, blood and tibia bone. The MOLP supplementation significantly increased pH of breast muscle and ash percentage of tibia bone. The diameter of breast muscle fibres and also weight and weight length index of tibia bone significantly increased with 12 and 15 g/kg MOLP. The water holding capacity (WHC) of breast muscle was significantly higher with 9 and 15 g/kg MOLP; whereas robusticity index of tibia bone significantly decreased with 12 and 15 g/kg MOLP inclusion compared to the control group. In conclusion, dietary supplementation of Moringa oleifera leaf powder (12g/kg) increased pH, water holding capacity, and muscle fibre diameter of breast muscle and also weight, ash percentage and density indices of tibia bone in broiler chickens.

RESUMO: O grão-de-bico é a terceira cultura mais importante das áreas de sequeiro do Paquistão, mas é gravemente afetado pela planta daninha nociva Asphodelus tenuifolius. Por esse motivo, foi realizado um experimento de campo para avaliar o efeito de diferentes extratos alelopáticos de plantas daninhas, herbicidas e coberturas vegetais no controle de A. tenuifolius e outras plantas daninhas associadas ao grão-de-bico no distrito de Karak, na província de Khyber Pakhtunkhwa, Paquistão. O delineamento experimental utilizado foi em blocos casualizados (DBC) com três repetições, usando o cultivar de grão-de-bico Chattan, cultivado para o experimento. Foram utilizados neste estudo nove tratamentos, juntamente com o controle, para fins de comparação: Stomp 330 CE (pré-emergência) a 2,5 L ha-1 , fenoxaprop-p-ethyl, bromoxanil + MCPA (mistura em tanque), Starane-M, cobertura vegetal (folhas de eucalipto), cobertura vegetal (palha de trigo), extrato alelopático de planta daninha (A. tenuifolius) + Stomp, extrato alelopático de planta daninha (Cyperus rotundus) e extrato alelopático de planta daninha (Sorghum halepense), e o tratamento controle. Foram registrados dados sobre a densidade m-2 de A. tenuifolius antes e depois da aplicação das práticas de manejo de plantas daninhas (m-2), peso fresco de A. tenuifolius (m-2), rendimento de sementes (kg ha-1), rendimento biológico (kg ha-1) e relação custo-benefício. A densidade de plantas daninhas antes do manejo foi considerada não significativa. Os dados mostraram que a densidade mínima deA. tenuifolius após o manejo de plantas daninhas e o peso fresco (m-2) foram observados com Stomp 330 EC (6,33 m-2 e 1,98 kg m-2), que é estatisticamente semelhante ao fenoxaprop-p-ethyl 6,9 EC (10,33 m-2 e 3,03 kg m-2), enquanto a densidade máxima de A. tenuifolius e o peso fresco (m-2) foram registrados para as parcelas de controle (74,33 m-2 e 287,46 kg m-2). Os maiores rendimento máximo de sementes (1.781,7 kg ha-1), rendimento biológico (3.823 kg ha-1) e relação custo-benefício (3,47) foram registrados para as parcelas tratadas com Stomp 330 EC, ao passo que os menores rendimento de sementes (851,7 kg ha-1), rendimento biológico (3.126,3 kg ha-1) e relação custo-benefício (1,53) foram observados para a parcela de controle. Entre os parâmetros nutritivos do grão-de-bico, os teores máximos de proteína bruta (17,40%), gordura bruta (4,90%) e óleo (5,98%) foram registrados para parcelas tratadas com Stomp 330 EC, enquanto os menores teores de proteína bruta (16,18%), gordura bruta (4,12%) e óleo (5,01%) foram encontrados nas parcelas de controle. Assim, recomenda-se que os herbicidas Stomp 330 EC e fenoxaprop-p-ethyl sejam utilizados em pré-emergência nas doses recomendadas para o controle de A. tenuifolius e outras plantas daninhas associadas em culturas de grão-de-bico.

ABSTRACT: Chickpea is the third most important crop of rainfed areas of Pakistan, and it is severely affected by noxious weed Asphodelus tenuifolius. Therefore, a field trial was conducted to evaluate the effect of different allelopathic weed extracts, herbicides and mulches in controlling A. tenuifolius and other associated weeds in chickpea at “District Karak” Khyber Pakhtunkhwa-Pakistan. The experiment was carried out in a Randomized Complete Block Design (RCBD) and replicated thrice using a chickpea cultivar (Chattan) which was grown for the experimental trial. There were 9 treatments along with control for comparison. The treatments Stomp 330 EC (Pre-emergence) at 2.5 L ha-1, Fenoxaprop-p-ethyl, Bromoxanil+MCPA (Tank mix), Starane-M, mulching (Eucalyptus leaves), mulching (Wheat straw mulch), allelopathic weed extract (A. tenuifolius) + Stomp, allelopathic weed extract (Cyperus rotundus), allelopathic weed extract (Sorghum halepense) and control treatment were used in this trial. Data were recorded on A. tenuifolius density m-2 before and after application of weed management practices (m-2), A.tenuifolius fresh weight (m-2), seed yield (kg.ha-1), biological yield (kg ha-1) and cost-benefit ratio. Weed density before weed management was found to be non-significant. The data showed that minimum A. tenuifolius density after weed management and fresh weight (m-2) were found for Stomp 330 EC (6.33 m-2 and 1.98 kg m-2), which is statistically similar to Fenoxaprop-p-ethyl 6.9 EC (10.33 m-2 and 3.03 kg m-2) while maximum A. tenuifolius density and fresh weight (m-2) were recorded for control plots (74.33 m-2 and 287.46 kg m-2). Maximum seed yield (1,781.7 kg ha-1), biological yield (3,823 kg ha-1) and cost-benefit ratio (3.47) were recorded for plots treated with Stomp 330 EC, while the lowest seed yield (851.7 kg ha-1), biological yield (3,126.3 kg ha-1) and cost-benefit ratio (1.53) were observed for the control plot. Among the nutritive parameters in chickpea, maximum crude protein (17.40%), crude fat content (4.90%) and oil content (5.98%) were recorded for plots treated with Stomp 330 EC, while minimum crude protein (16.18%), crude fat content (4.12%) and oil content (5.01%) were found in the control plots. Thus, it is recommended that herbicides Stomp 330 EC and Fenoxaprop-p-ethyl should be used at pre-emergence at their recommended doses for control of A.tenuifolius and other associated weeds in chickpea crops.

The dietary supplementation of trace minerals and probiotics has long been practiced to reduce the harmful effects of high environmental temperature in poultry. The study aimed to assess the effect of single or combined supplementation of ZnSO4●H2O and probiotic (Protexin®) on the growth performance, gut biometry and immune organs histomorphology in broilers reared under cyclic heat stress. For this purpose, 280 day-old broiler chicks were randomly divided into seven groups designated as control (basal diet), HS (heat stress + basal diet), HS-Zn30 (HS + 30 mg ZnSO4●H2O/kg feed), HS-Zn60 (HS + 60 mg ZnSO4●H2O/kg feed), HS-Pro (HS + 0.1 g Protexin®/kg feed), HS-Zn30-Pro (HS + 30 mg ZnSO4●H2O/kg feed + 0.1 g Protexin®/kg feed), and HS-Zn60-Pro (HS + 60 mg ZnSO4●H2O/kg feed + 0.1 g Protexin®/kg feed). Each group consisted of five replicates (8 birds per replicate). The birds were fed on starter (0 - 21days) and grower (22 - 42 days) diets. From day 22 onwards, the birds were exposed to cyclic heat stress (temperature and relative humidity at 35 ± 1 °C and 75 ± 5% respectively) for 8 hours. The results revealed that birds exposed to heat stress showed significant decrease in body weight (BW) and increase in feed conversion ratio (FCR) compared to the control group. The dietary supplementation of ZnSO4●H2O and probiotic either singly or in combination improved BW and FCR throughout the experimental period in heat-stressed broilers. Area of lymphatic nodule and lymphoid follicle in caecal tonsils and bursa of Fabricius increased in HS-Pro, HS-Zn30-Pro and HS-Zn60-Pro groups compared to HS group. In conclusion, zinc and probiotics combination partially improved performance and immune organs histomorphology in heat-stressed broilers.