The plant glycosyltransferase clone collection for functional genomics.

The glycosyltransferases (GTs) are an important and functionally diverse family of enzymes involved in glycan and glycoside biosynthesis. Plants have evolved large families of GTs which undertake the array of glycosylation reactions that occur during plant development and growth. Based on the Carbohydrate-Active enZymes (CAZy) database, the genome of the reference plant Arabidopsis thaliana codes for over 450 GTs, while the rice genome (Oryza sativa) contains over 600 members. Collectively, GTs from these reference plants can be classified into over 40 distinct GT families. Although these enzymes are involved in many important plant specific processes such as cell-wall and secondary metabolite biosynthesis, few have been functionally characterized. We have sought to develop a plant GTs clone resource that will enable functional genomic approaches to be undertaken by the plant research community. In total, 403 (88%) of CAZy defined Arabidopsis GTs have been cloned, while 96 (15%) of the GTs coded by rice have been cloned. The collection resulted in the update of a number of Arabidopsis GT gene models. The clones represent full-length coding sequences without termination codons and are Gateway® compatible. To demonstrate the utility of this JBEI GT Collection, a set of efficient particle bombardment plasmids (pBullet) was also constructed with markers for the endomembrane. The utility of the pBullet collection was demonstrated by localizing all members of the Arabidopsis GT14 family to the Golgi apparatus or the endoplasmic reticulum (ER). Updates to these resources are available at the JBEI GT Collection website http://www.addgene.org/.

A ß-glucuronosyltransferase from Arabidopsis thaliana involved in biosynthesis of type II arabinogalactan has a role in cell elongation during seedling growth.

We have characterized a ß-glucuronosyltransferase (AtGlcAT14A) from Arabidopsis thaliana that is involved in the biosynthesis of type II arabinogalactan (AG). This enzyme belongs to the Carbohydrate Active Enzyme database glycosyltransferase family 14 (GT14). The protein was localized to the Golgi apparatus when transiently expressed in Nicotiana benthamiana. The soluble catalytic domain expressed in Pichia pastoris transferred glucuronic acid (GlcA) to ß-1,6-galactooligosaccharides with degrees of polymerization (DP) ranging from 3-11, and to ß-1,3-galactooligosaccharides of DP5 and 7, indicating that the enzyme is a glucuronosyltransferase that modifies both the ß-1,6- and ß-1,3-galactan present in type II AG. Two allelic T-DNA insertion mutant lines showed 20-35% enhanced cell elongation during seedling growth compared to wild-type. Analyses of AG isolated from the mutants revealed a reduction of GlcA substitution on Gal-ß-1,6-Gal and ß-1,3-Gal, indicating an in vivo role of AtGlcAT14A in synthesis of those structures in type II AG. Moreover, a relative increase in the levels of 3-, 6- and 3,6-linked galactose (Gal) and reduced levels of 3-, 2- and 2,5-linked arabinose (Ara) were seen, suggesting that the mutation in AtGlcAT14A results in a relative increase of the longer and branched ß-1,3- and ß-1,6-galactans. This increase of galactosylation in the mutants is most likely caused by increased availability of the O6 position of Gal, which is a shared acceptor site for AtGlcAT14A and galactosyltransferases in synthesis of type II AG, and thus addition of GlcA may terminate Gal chain extension. We discuss a role for the glucuronosyltransferase in the biosynthesis of type II AG, with a biological role during seedling growth.

Three members of the Arabidopsis glycosyltransferase family 8 are xylan glucuronosyltransferases.

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 ß(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.

Isolation and proteomic characterization of the Arabidopsis Golgi defines functional and novel components involved in plant cell wall biosynthesis.

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 thaliana) 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.

Glycoside hydrolases from a targeted compost metagenome, activity-screening and functional characterization.

BACKGROUND: Metagenomics approaches provide access to environmental genetic diversity for biotechnology applications, enabling the discovery of new enzymes and pathways for numerous catalytic processes. Discovery of new glycoside hydrolases with improved biocatalytic properties for the efficient conversion of lignocellulosic material to biofuels is a critical challenge in the development of economically viable routes from biomass to fuels and chemicals. RESULTS: Twenty-two putative ORFs (open reading frames) were identified from a switchgrass-adapted compost community based on sequence homology to related gene families. These ORFs were expressed in E. coli and assayed for predicted activities. Seven of the ORFs were demonstrated to encode active enzymes, encompassing five classes of hemicellulases. Four enzymes were over expressed in vivo, purified to homogeneity and subjected to detailed biochemical characterization. Their pH optima ranged between 5.5 - 7.5 and they exhibit moderate thermostability up to ~60-70°C. CONCLUSIONS: Seven active enzymes were identified from this set of ORFs comprising five different hemicellulose activities. These enzymes have been shown to have useful properties, such as moderate thermal stability and broad pH optima, and may serve as the starting points for future protein engineering towards the goal of developing efficient enzyme cocktails for biomass degradation under diverse process conditions.

Engineering microbial biofuel tolerance and export using efflux pumps.

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.

An investigation on protein and amino acid contents in scales and muscles of pomfret Parastromateus niger (Bloch, 1795) and Pampus argenteus (Eupharasen, 1788).

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 these Pomfret 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.

Assessments of some trace metals in water samples of nursery pond of Grass Carp (Ctenopharyngodon idella, Valenciennes, 1844) in Bannu Fish Hatchery of Khyber Pakhtunkhwa, Pakistan.

The present investigation was aimed to examine the concentrations of trace metals including e.g copper (Cu), manganese (Mn), nickel (Ni), and zinc (Zn) in water samples collected from nursery pond of grass carp (Ctenopharyngodon idella) in Bannu Hatchery of Khyber Pakhtunkhwa during the period from April 2018 to January 2019. The temperature and pH of each water sample were measured for the whole study duration. The concentration of Copper (Cu), Manganese (Mn), Nickel (Ni), and Zinc (Zn) in collected water samples were measured in mg/liter by using Atomic Absorption Spectrophotometer. The blank and standard solutions for device calibration Standard solutions i.e., 2.0 mg, 4.0 mg, and 6.0 were used to measure the concentration of these metals in water samples to verify the measurements. The data was statistically analyzed on descriptive statistics (estimation of proportions and standard deviation) used to summarize mean concentration. The results obtained of both temperature and pH of water samples were found in ranged 10 to 36 0C and 7.0 to 8.44; whereas the size of fry stages was ranged from 4.0 to 56.0 mm in total length. The results of investigated metals found in pond water samples are in order of Zn>Mn>Ni>Cu, respectively. As optimum temperature and pH for grass carp were mostly between 15 0C and 30 0C and pH 6.5 to 8.0. It was concluded from obtained results that temperature, pH, and trace metals were found appropriate for the growth of Ctenopharyngodon idella from fry to fingerling stages, but the highest amount of zinc can cause its mortality. It is a preliminary study on grass carp culturing in Bannu fish hatchery so, it would provide useful information for model fish seed production unit in a hatchery.

Monitoring pond water quality to improve the production of Labeo rohita (Hamilton, 1822) in Bannu Fish Hatchery of Bannu district, Khyber Pakhtunkhwa province; An Implications for artificial fish culture.

The water quality parameters of a fish pond are essential to be managing properly under control for successful operations of fish culture. Improper management of pond water quality during the juvenile stages can create stressful conditions to produce various harmful diseases, which may decrease the fish quality and results in low profits. The present study was aimed to monitored important water quality parameters of nursery ponds of Labeo rohita culturing in Bannu fish hatchery. The study duration was 75 days extends from 10th June to 24th August 2019 for the successful culture of this specie which can play a significant role in breeding season. Furthermore, the concentration of some heavy metals like copper (Cu), nickel (Ni), manganese (Mn), iron (Fe), cadmium (Cd), and zinc (Zn) in pond water and fry stages of this species was also determined. The data obtained from all water quality parameters were analyzed expressed as range, mean and standard deviation using MS Excel 2013. The obtained results of 75 days study revealed that the water pH & temperature, electric conductivity, total dissolved solids (TDS), and total dissolved oxygen (DO) of pond water samples were found within a tolerable limit except salinity and dissolved ammonia concentration were not permissible for fish growth. The average concentration of heavy metals in pond water exhibited descending order Fe>Ni>Mn>Zn>Cd>Ni, which was found in acceptable ranges. Whereas, the average values of heavy metals in fry stages were in the order of Zn>Fe>Ni>Cu>Cd>Mn, and found within the recommended values of WHO/FAO. Thus, it was concluded from this study that good water quality is a precondition, maintaining balanced levels of water quality parameters is fundamental for both the health and growth of fish culture which is quite necessary for assuring increased fish productivity. It is recommended to monitor and assess water quality parameters on a routine basis for promoting healthy fish culture.

Prevalence of intestinal nematodes infection in school children of urban areas of district Lower Dir, Pakistan.

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.

Study of some morphometric and meristic characteristics of Alepes vari (Cuvier, 1833) collected from the Arabian coast.

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.

Growth performance and survivability of the Asian seabass Lates calcarifer (Bloch, 1790) reared under hyper-saline, hypo-saline and freshwater environments in a closed aquaculture system.

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.

Bioinformatic processing to identify single nucleotide polymorphism that potentially affect Ape1 function.

Inactivation of DNA damage response mechanisms is associated with several disease syndromes, including cancer, aging and neurodegeneration. A major corrective pathway for alkylation or oxidative DNA damage is base excision repair (BER). As part of an effort to identify variation in DNA repair genes, we used the expressed sequence tag (EST) database to identify amino acid variation in Ape1, an essential gene in the BER repair pathway. Nucleotide substitutions were considered valid only if the amino acid changes were observed in at least two independent EST sequencing runs (i.e. two independent EST reports). In total eighty amino acid variants were identified for the Ape1 gene. Using software tools SIFT and PolyPhen, which predict impacts of amino acid substitutions on protein structure and function, twenty-six variants were predicted by both algorithms to be deleterious to protein function. Majority of these intolerant mutations such as V206C and F240S, lie within the core of the protein and may affect the stability and folding of Ape1, or in the case of N212H, N212K, and Y171N, are close to the enzyme's active site and could drastically affect its function. A few of the intolerant mutations, i.e., G178V and E217R, are surface residues and are far from the active site, and as such, the predicted effect on Ape1 stability or function is not evident. These variants are reagents for further protein function studies and molecular epidemiology studies of cancer susceptibility.

Production of tranilast [N-(3',4'-dimethoxycinnamoyl)-anthranilic acid] and its analogs in yeast Saccharomyces cerevisiae.

Biological synthesis of therapeutic drugs beneficial for human health using microbes offers an alternative production strategy to the methods that are commonly employed such as direct extraction from source organisms or chemical synthesis. In this study, we evaluated the potential for yeast (Saccharomyces cerevisiae) to be used as a catalyst for the synthesis of tranilast and various tranilast analogs (cinnamoyl anthranilates). Several studies have demonstrated that these phenolic amides have antioxidant properties and potential therapeutic benefits including antiinflammatory, antiproliferative, and antigenotoxic effects. The few cinnamoyl anthranilates naturally produced in plants such as oats and carnations result from the coupling of various hydroxycinnamoyl-CoAs to anthranilic acid. In order to achieve the microbial production of tranilast and several of its analogs, we engineered a yeast strain to co-express a 4-coumarate/CoA ligase (4CL, EC 6.2.1.12) from Arabidopsis thaliana and a hydroxycinnamoyl/benzoyl-CoA/anthranilate N-hydroxycinnamoyl/benzoyltransferase (HCBT, EC 2.3.1.144) from Dianthus caryophyllus. This modified yeast strain allowed us to produce tranilast and 26 different cinnamoyl anthranilate molecules within a few hours after exogenous supply of various combinations of cinnamic acids and anthranilate derivatives. Our data demonstrate the feasibility of rapidly producing a wide range of defined cinnamoyl anthranilates in yeast and underline a potential for the biological designed synthesis of naturally and non-naturally occurring molecules.

Impact of dietary leaf extracts of Black pepper Piper nigrum L. on the growth, hematological and immunological parameters of Labeo rohita (Hamilton, 1822) cultured in glass aquaria.

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.

Comparative analysis of brain in relation to the body length and weight of common carp (Cyprinus carpio) in captive (hatchery) and wild (river system) populations.

Cyprinus carpio is the member of family cyprinidae commonly called common carp. This study was aimed to find out the comparison of brain of wild (river system) and captive (hatchery reared) population of common carp. A total of thirty samples (15 from hatchery and 15 from river Swat) were collected. All the specimens were examined in Laboratory of Parasitoloy, Zoology Department, University of Malakand. Findings indicated that wild population were greater in brain size and weight as compared to hatchery reared population. The fish samples collected from captive environment (hatchery) were showing more weight and length as compared to wild population of common carps. The mean value of total weight of hatchery fishes 345±48.68 and the mean value of brain weight of hatchery reared fishes 0.28±0.047. The mean value of wild fish's total body weight 195.16±52.58 and the mean value of brain weight of wild fishes are 0.45±0.14. Present research calls for the fact that fish in dependent environmental conditions possess brain larger in size as compared to its captive population, it is due to use and disuse of brain in their environmental requirements.

Evaluation of indigenous plants' extracts for mosquitocidal activity against different stages of Culex quinquefasciatus say (Diptera: Culicidae).

Being vector of West Nile Virus and falariasis the control of Culex quinquefasciatus is likely to be essential. Synthetic insecticide treatment is looking most effective for vectors mosquito control. However, these products are toxic to the environment and non-target organisms. Consequently, ecofriendly control of vectors mosquito is needed. In this regard botanical insecticide is looking more fruitful. Therefore, the present research aimed to investigate the effectiveness of methanolic extract and various fractions, including, n-hexane, ethyl-acetate, chloroform, and aqueous fraction, obtained from methanolic extract of Ailanthus altissima, Artemisia scoparia, and Justicia adhatoda using separating funnel against larval, pupal, and adult stages of Culex quinquefasciatus. The larvae and pupae of Culex quinquefasciatus were exposed to various concentrations (31.25-1000 ppm) of methanolic extract and its fractions for 24 hours of exposure period. For knock-down bioassay (filter paper impregnation bioassay) different concentration of the methanolic extract and its various fractions (i.e. 0.0625, 0.125, 0.25, 0.5 and 1mg/mL) were applied for 1 hour exposure period. The results were statistically analysed using standard deviation, probit analysis, and linear regression. The R2 values of larvae, pupae, and adult range from 0.4 to 0.99. The values of LC50 (concentration causing 50% mortality) for late 3rd instar larvae after 24 hours exposure period range from 93-1856.7 ppm, while LC90 values range from 424 -7635.5ppm. The values of LC50for pupae range form 1326.7-6818.4ppm and and values of LC90 range from 3667.3-17427.9ppm, respectively. The KDT50 range from 0.30 to 2.8% and KDT90 values range from1.2 to 110.8%, respectively. In conclusion, Justicia adhatoda may be effective for controlling populations of vector mosquito.

Prevalence of cutaneous leishmaniasis in the largest populated city Karachi, Pakistan.

The study was undertaken from September 2018 to April 2019 to determine the prevalence of cutaneous leishmaniasis in human beings living in six districts of Karachi. Suspected persons were screened for the disease and positive cases were identified on the basis of skin lesions and blood samples. Samples were observed by mounting their smear. A total of 207 subjects of different ages and sex groups were investigated, however, only 192 (92%) of the suspected cases were found to have the disease 64% of cases were male which were significantly high (p 0.05), than female 36%. The lesion was more frequently detected among the youth ages of 21-30 years (31%) as compared to other groups. In both sexes, legs were found to be more infected (25% male + 20% female) followed by arms (20% male + 0% female) and face (15% male +11% female). The mixed body parts had shown the lowest infections such as (4% in males + 5%) in females. In conclusion, the highest and lowest leishmaniasis infections were observed in District West (23% male + 9% female) followed by District East (15% male + 7% female), District Malir (11% male+ 4% female), District Central (7% male + 5% female), District Korangi (4% male + 7% female) and District South (4% male + 4% female) respectively.

Biodiversity of commercially important finfish species caught by mid-water and bottom trawls from two different coasts of Arabian Sea: Threats and conservation strategies.

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.

Prevalence of intestinal parasitic diseases in school children of rural areas of district Lower Dir, Pakistan.

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.