DNA bar-coding is a taxonomic method that uses small genetic markers in organisms' mitochondrial ... more DNA bar-coding is a taxonomic method that uses small genetic markers in organisms' mitochondrial DNA (mt DNA) for identification of particular species. It uses sequence diversity in a 658-base pair fragment near the 5 0 end of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene as a tool for species identification. DNA barcoding is more accurate and reliable method as compared with the morphological identification. It is equally useful in juveniles as well as adult stages of fishes. The present study was conducted to identify three farm fish species of Pakistan (Cyprinus carpio, Cirrhinus mrigala, and Ctenopharyngodon idella) genetically. All of them belonged to family cyprinidae. CO1 gene was amplified. PCR products were sequenced and analyzed by bioinformatic software. Conspecific, congenric, and confamilial k2P nucleotide divergence was estimated. From these findings, it was concluded that the gene sequence, CO1, may serve as milestone for the identification of related species at molecular level.
Fish taxonomy plays a fundamental role in the study of biodiversity. However, traditional methods... more Fish taxonomy plays a fundamental role in the study of biodiversity. However, traditional methods of fish taxonomy rely on morphological features, which can lead to confusion due to great similarities between closely related species. To overcome this limitation, modern taxonomy employs DNA barcoding as a species identification method. This involves using a short standardized mitochondrial DNA region as a barcode, specifically a 658 base pair fragment near the 5′ end of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene, to exploit diversity in this region for identification of species. To test the effectiveness and reliability of DNA barcoding, 25 fish specimens from nine different fish species found in various rivers of Pakistan were identified morphologically using a dichotomous key at the start of the study. comprising nine fresh water fish species including Mystus cavasius, Mystus bleekeri, Osteobrama cotio, Labeo rohita, Labeo culbasu, Labeo gonius, Cyprinus carpio, Catla catla and Cirrhinus mrigala from different rivers of Pakistan were used in present study. DNA was extracted from one of the pectoral fins and partial sequence of CO1 gene was amplified using conventional PCR method. Analysis of the barcodes confirmed that genetically identified fishes were the same as those identified morphologically at the beginning of the study. The sequences were also analyzed for biodiversity and phylogenetic studies. Based on the results of the study, it can be concluded that DNA barcoding is an effective and reliable method for studying biodiversity and conducting phylogenetic analysis of different fish species in Pakistan.
DNA barcoding technique involving documentation of animal species based on a partial sequence of ... more DNA barcoding technique involving documentation of animal species based on a partial sequence of cytochrome C Oxidase 1 gene from mitochondrial DNA attracted many scientists at the start of 21st century. The flaws in morphometric methods for identification of larvae of fishes and other animals, their eggs, and damaged specimen with incomplete morphology upraised the question for an alternate method for identification and grouping of many animal species. Partial sequence of CO1 gene used as barcode has been attested as a valuable tool for identification of fish species as well as supportive in computing evolutionary history and genetic diversity. Current study was conducted to identify the fishes by using the DNA barcoding technique resulting partial sequences of CO1 gene, and then use of these sequences in evaluation of the evolutionary history and genetic diversity of Cirrhinus mrigala inhabiting different areas. Short sequences from 5 end of CO1 gene (650 base pair) were amplified, sequenced and analyzed using different soft wares. According to results haplotypes were found which showed genetic variations among fishes from two different habitats. The phylogenetic analysis established a neighboring relationship and a shared ancestor of fish from different habitats. Haplotype diversity of 0.7143 was found in partial sequences of CO1 gene of all experimental fish. From these results it is obvious that fish species sharing identical genus and family but with diverse habitats show genetic variations even though involving a common ancestor.
The cow is a very important mammal from the bovine family renowned for providing milk, meat and h... more The cow is a very important mammal from the bovine family renowned for providing milk, meat and hide around the globe. β-casein is one of the principal proteins found in cow’s milk encoded by the CSN2 gene with 12-13 reported genetic variants present on Chr.6 in the cow’s genome. The current study investigated A1/A2 associated variant g.6:85451298C>A (rs43703011) in exon 7th according to the GenBank ID: M55158.1 position 8101C>A (c.245C>A), which is responsible to change the 67th amino acid of the β-casein polypeptide from proline (CCT, A2) to histidine (CAT, A1). The milk containing A2 protein variant is good while A1 milk has some negative effects on human health due to the production of β-casmomorphin-7. Genetic screening of the subject variant was conducted using ARMS-PCR on 48 native and exotic cows from Pakistan. Our initial results showed that 37.5% of the population is homozygous wild-type (C/C), 56.25% is heterozygous (C/A) and the remaining 6.25% is homozygous mut...
Deltamethrin is a broad spectrum synthetic pyrethroid pesticide. It is considered less toxic as c... more Deltamethrin is a broad spectrum synthetic pyrethroid pesticide. It is considered less toxic as compared to organochlorine and organo-phosphate pesticides. Deltamethrin was selected to study the biochemical and histological alterations in liver of silver carp. The LC50 1.6 μg/L was determined. Fish (n=6) were dividing into 4 groups A (control) B, C and D (experimental). Experimental groups B, C and D were exposed to 25%, 50% and 75% of LC50 respectively for acute exposure. The blood and liver tissue sampling was done after 96hrs for histology and biochemical analysis [alanine amino transferase (ALT), and aspartate amino-transferase (AST)]. Liver histology revealed that necrosis, nuclear pycnosis, hypertrophy of hepatocytes, vacuolization, nuclear atrophy and congestion of blood vessels were observed as compare to control group. This result was also supported by the significant increase in levels of hepatic enzymes AST and ALT in blood plasma of exposed fish as compared to control fi...
DNA barcoding technique involving documentation of animal species based on a partial sequence of ... more DNA barcoding technique involving documentation of animal species based on a partial sequence of cytochrome C Oxidase 1 gene from mitochondrial DNA attracted many scientists at the start of 21 century. The flaws in morphometric methods for identification of larvae of fishes and other animals, their eggs, and damaged specimen with incomplete morphology upraised the question for an alternate method for identification and grouping of many animal species. Partial sequence of CO1 gene used as barcode has been attested as a valuable tool for identification of fish species as well as supportive in computing evolutionary history and genetic diversity. Current study was conducted to identify the fishes by using the DNA barcoding technique resulting partial sequences of CO1 gene, and then use of these sequences in evaluation of the evolutionary history and genetic diversity of Cirrhinus mrigala inhabiting different areas. Short sequences from 5 end of CO1 gene (650 base pair) were amplified, ...
Organisms have naturally build defense mechanisms against infections and diseases caused by patho... more Organisms have naturally build defense mechanisms against infections and diseases caused by pathogens. This system known as immune system comprises of innate immunity and adaptive immunity. Innate immunity is non specific and produces a quick response to the invading pathogens. Macrophages play central role in innate immunity, also called natural immunity. The other part of immunity is rather specific and it takes some time to show its response, this type of immunity is called adaptive immunity. Adaptive immunity can be either T cell mediated or B cell mediated in response to intra cellular or extra cellular pathogens respectively. When it is T cell mediated it will lead to the secretion of cytokines including interferon along with other substances, which help to fight intra cellular pathogens especially viruses by activating natural killer cells and macrophages. B cell mediated immune system leads to production of antibodies in response to extra cellular pathogens in blood or other...
DNA bar-coding is a taxonomic method that uses small genetic markers in organisms&amp... more DNA bar-coding is a taxonomic method that uses small genetic markers in organisms' mitochondrial DNA (mt DNA) for identification of particular species. It uses sequence diversity in a 658-base pair fragment near the 5' end of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene as a tool for species identification. DNA barcoding is more accurate and reliable method as compared with the morphological identification. It is equally useful in juveniles as well as adult stages of fishes. The present study was conducted to identify three farm fish species of Pakistan (Cyprinus carpio, Cirrhinus mrigala, and Ctenopharyngodon idella) genetically. All of them belonged to family cyprinidae. CO1 gene was amplified. PCR products were sequenced and analyzed by bioinformatic software. Conspecific, congenric, and confamilial k2P nucleotide divergence was estimated. From these findings, it was concluded that the gene sequence, CO1, may serve as milestone for the identification of related species at molecular level.
Taxonomy has always been a challenging job for the scientists. Traditional methods for identifica... more Taxonomy has always been a challenging job for the scientists. Traditional methods for identification of which sometimes create great confusion due to great morphological similarities in closed species. DNA barcoding is a modern species identification method in which a short standardized mitochondrial DNA region is u fragment near the 5 (CO1 capable enough for identification of processed, fishes as compared to traditional morphometric methods and is also appropriate for rapid identification of juveniles as well as adult sta fishes. Present study is Labeogonius of CO1 and same fishes which were identified morphologically at the b study with the help of identification key. The phylogenetic tree analysis confirmed the evolutionary relationships between from Pakistan and fishes from other parts of the world. From these findings, it is marker for identification of most of fish species at molecular phylogenetic analysis.
DNA bar-coding is a taxonomic method that uses small genetic markers in organisms' mitochondrial ... more DNA bar-coding is a taxonomic method that uses small genetic markers in organisms' mitochondrial DNA (mt DNA) for identification of particular species. It uses sequence diversity in a 658-base pair fragment near the 5 0 end of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene as a tool for species identification. DNA barcoding is more accurate and reliable method as compared with the morphological identification. It is equally useful in juveniles as well as adult stages of fishes. The present study was conducted to identify three farm fish species of Pakistan (Cyprinus carpio, Cirrhinus mrigala, and Ctenopharyngodon idella) genetically. All of them belonged to family cyprinidae. CO1 gene was amplified. PCR products were sequenced and analyzed by bioinformatic software. Conspecific, congenric, and confamilial k2P nucleotide divergence was estimated. From these findings, it was concluded that the gene sequence, CO1, may serve as milestone for the identification of related species at molecular level.
Fish taxonomy plays a fundamental role in the study of biodiversity. However, traditional methods... more Fish taxonomy plays a fundamental role in the study of biodiversity. However, traditional methods of fish taxonomy rely on morphological features, which can lead to confusion due to great similarities between closely related species. To overcome this limitation, modern taxonomy employs DNA barcoding as a species identification method. This involves using a short standardized mitochondrial DNA region as a barcode, specifically a 658 base pair fragment near the 5′ end of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene, to exploit diversity in this region for identification of species. To test the effectiveness and reliability of DNA barcoding, 25 fish specimens from nine different fish species found in various rivers of Pakistan were identified morphologically using a dichotomous key at the start of the study. comprising nine fresh water fish species including Mystus cavasius, Mystus bleekeri, Osteobrama cotio, Labeo rohita, Labeo culbasu, Labeo gonius, Cyprinus carpio, Catla catla and Cirrhinus mrigala from different rivers of Pakistan were used in present study. DNA was extracted from one of the pectoral fins and partial sequence of CO1 gene was amplified using conventional PCR method. Analysis of the barcodes confirmed that genetically identified fishes were the same as those identified morphologically at the beginning of the study. The sequences were also analyzed for biodiversity and phylogenetic studies. Based on the results of the study, it can be concluded that DNA barcoding is an effective and reliable method for studying biodiversity and conducting phylogenetic analysis of different fish species in Pakistan.
DNA barcoding technique involving documentation of animal species based on a partial sequence of ... more DNA barcoding technique involving documentation of animal species based on a partial sequence of cytochrome C Oxidase 1 gene from mitochondrial DNA attracted many scientists at the start of 21st century. The flaws in morphometric methods for identification of larvae of fishes and other animals, their eggs, and damaged specimen with incomplete morphology upraised the question for an alternate method for identification and grouping of many animal species. Partial sequence of CO1 gene used as barcode has been attested as a valuable tool for identification of fish species as well as supportive in computing evolutionary history and genetic diversity. Current study was conducted to identify the fishes by using the DNA barcoding technique resulting partial sequences of CO1 gene, and then use of these sequences in evaluation of the evolutionary history and genetic diversity of Cirrhinus mrigala inhabiting different areas. Short sequences from 5 end of CO1 gene (650 base pair) were amplified, sequenced and analyzed using different soft wares. According to results haplotypes were found which showed genetic variations among fishes from two different habitats. The phylogenetic analysis established a neighboring relationship and a shared ancestor of fish from different habitats. Haplotype diversity of 0.7143 was found in partial sequences of CO1 gene of all experimental fish. From these results it is obvious that fish species sharing identical genus and family but with diverse habitats show genetic variations even though involving a common ancestor.
The cow is a very important mammal from the bovine family renowned for providing milk, meat and h... more The cow is a very important mammal from the bovine family renowned for providing milk, meat and hide around the globe. β-casein is one of the principal proteins found in cow’s milk encoded by the CSN2 gene with 12-13 reported genetic variants present on Chr.6 in the cow’s genome. The current study investigated A1/A2 associated variant g.6:85451298C>A (rs43703011) in exon 7th according to the GenBank ID: M55158.1 position 8101C>A (c.245C>A), which is responsible to change the 67th amino acid of the β-casein polypeptide from proline (CCT, A2) to histidine (CAT, A1). The milk containing A2 protein variant is good while A1 milk has some negative effects on human health due to the production of β-casmomorphin-7. Genetic screening of the subject variant was conducted using ARMS-PCR on 48 native and exotic cows from Pakistan. Our initial results showed that 37.5% of the population is homozygous wild-type (C/C), 56.25% is heterozygous (C/A) and the remaining 6.25% is homozygous mut...
Deltamethrin is a broad spectrum synthetic pyrethroid pesticide. It is considered less toxic as c... more Deltamethrin is a broad spectrum synthetic pyrethroid pesticide. It is considered less toxic as compared to organochlorine and organo-phosphate pesticides. Deltamethrin was selected to study the biochemical and histological alterations in liver of silver carp. The LC50 1.6 μg/L was determined. Fish (n=6) were dividing into 4 groups A (control) B, C and D (experimental). Experimental groups B, C and D were exposed to 25%, 50% and 75% of LC50 respectively for acute exposure. The blood and liver tissue sampling was done after 96hrs for histology and biochemical analysis [alanine amino transferase (ALT), and aspartate amino-transferase (AST)]. Liver histology revealed that necrosis, nuclear pycnosis, hypertrophy of hepatocytes, vacuolization, nuclear atrophy and congestion of blood vessels were observed as compare to control group. This result was also supported by the significant increase in levels of hepatic enzymes AST and ALT in blood plasma of exposed fish as compared to control fi...
DNA barcoding technique involving documentation of animal species based on a partial sequence of ... more DNA barcoding technique involving documentation of animal species based on a partial sequence of cytochrome C Oxidase 1 gene from mitochondrial DNA attracted many scientists at the start of 21 century. The flaws in morphometric methods for identification of larvae of fishes and other animals, their eggs, and damaged specimen with incomplete morphology upraised the question for an alternate method for identification and grouping of many animal species. Partial sequence of CO1 gene used as barcode has been attested as a valuable tool for identification of fish species as well as supportive in computing evolutionary history and genetic diversity. Current study was conducted to identify the fishes by using the DNA barcoding technique resulting partial sequences of CO1 gene, and then use of these sequences in evaluation of the evolutionary history and genetic diversity of Cirrhinus mrigala inhabiting different areas. Short sequences from 5 end of CO1 gene (650 base pair) were amplified, ...
Organisms have naturally build defense mechanisms against infections and diseases caused by patho... more Organisms have naturally build defense mechanisms against infections and diseases caused by pathogens. This system known as immune system comprises of innate immunity and adaptive immunity. Innate immunity is non specific and produces a quick response to the invading pathogens. Macrophages play central role in innate immunity, also called natural immunity. The other part of immunity is rather specific and it takes some time to show its response, this type of immunity is called adaptive immunity. Adaptive immunity can be either T cell mediated or B cell mediated in response to intra cellular or extra cellular pathogens respectively. When it is T cell mediated it will lead to the secretion of cytokines including interferon along with other substances, which help to fight intra cellular pathogens especially viruses by activating natural killer cells and macrophages. B cell mediated immune system leads to production of antibodies in response to extra cellular pathogens in blood or other...
DNA bar-coding is a taxonomic method that uses small genetic markers in organisms&amp... more DNA bar-coding is a taxonomic method that uses small genetic markers in organisms' mitochondrial DNA (mt DNA) for identification of particular species. It uses sequence diversity in a 658-base pair fragment near the 5' end of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene as a tool for species identification. DNA barcoding is more accurate and reliable method as compared with the morphological identification. It is equally useful in juveniles as well as adult stages of fishes. The present study was conducted to identify three farm fish species of Pakistan (Cyprinus carpio, Cirrhinus mrigala, and Ctenopharyngodon idella) genetically. All of them belonged to family cyprinidae. CO1 gene was amplified. PCR products were sequenced and analyzed by bioinformatic software. Conspecific, congenric, and confamilial k2P nucleotide divergence was estimated. From these findings, it was concluded that the gene sequence, CO1, may serve as milestone for the identification of related species at molecular level.
Taxonomy has always been a challenging job for the scientists. Traditional methods for identifica... more Taxonomy has always been a challenging job for the scientists. Traditional methods for identification of which sometimes create great confusion due to great morphological similarities in closed species. DNA barcoding is a modern species identification method in which a short standardized mitochondrial DNA region is u fragment near the 5 (CO1 capable enough for identification of processed, fishes as compared to traditional morphometric methods and is also appropriate for rapid identification of juveniles as well as adult sta fishes. Present study is Labeogonius of CO1 and same fishes which were identified morphologically at the b study with the help of identification key. The phylogenetic tree analysis confirmed the evolutionary relationships between from Pakistan and fishes from other parts of the world. From these findings, it is marker for identification of most of fish species at molecular phylogenetic analysis.
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