1.
Studies On Genetic Diversity Of Labeo Rohita And Cirrhinus Mrigala By Using Molecular Markers In Punjab-Pakistan
by Fayyaz Rasool | Prof. Dr. Naureen Aziz Qureshi | Prof. Dr. Muhammad Ashraf.
Material type: ; Format:
print
Publisher: 2012Dissertation note: The studies on genetic diversity of Labeo rohita and Cirrhinus mrigala by using molecular markers in Punjab-Pakistan were carried out to investigate the genetic structure of said Indian major carps by RAPAD marker and the levels of polymorphism and similarity amongst the different groups of five populations of wild and farmed types. The results obtained from the present study after statistical analyses are presented in section-4 of this dissertation. The samples were collected from the following sites; for farmed fish was collected from UVAS-Fish Hatchery, C-block Ravi campus Pattoki district Kasur and for wild fish; from Trimu Barrage at the junction of River Chenab and Jhelum near district Jhang, Taunsa Barrage at River Indus near tehsil Kot Adu district Muzaffar Garh, Qadirabad Barrage at River Chenab near district Mandi Bahuddin and Baloki Barrage at River Ravi near tehsil Bhai Phero district Kasur. Analysis of variance (ANOVA) for the different morphometric parameters of study and Pearson's correlation among the physico-chemical parameters of water quality was done by Minitab statistical computer software. The XLSTAT 2012 version 1.02 of the computer software was used for the Pearson correlation analysis of the morphometric parameters of study. The same computer program was used for Agglomerative Hierarchical Clustering (AHC) of the different genotype occurrence on the basis of differences in morphometric parameters was done by Agglomeration method by following the Unweighted Pair Group Method with Arithmetic Mean (UPGMA). The Principle Component Analysis (PCA) on the basis of differentiation in morphometric parameters by Eigenvalues and differentiation into factors of the different genotypes from the different environmental conditions was done by correlation bi-plot/coefficient of the correlation (n) method in the same program. This software was also used to analyze the RAPAD data for Jaccard's coefficient by following the Unweighted Pair Group Method with Arithmetic Mean (UPGMA) for Hierarchical Clustering of the similar groups on the basis of similarity amongst the genotypes and the dendrogram generated is presented in the next section. The Principal Component Analysis (PCA) for grouping of the different genotypes from the different environmental conditions was done by Spearman Varimax rotation method for bi-plot generation of the co-occurrence of the same genotypes with similar genetic properties and specificity of different primersin the same program.
Following results were obtained:
Morphometric Parameters
1. Morphometric parameters of L. rohitashowed following trends: body weight, total length and average length of paired pectoral fins were nonsignificantly different (p > 0.05), fork length, dorsal fin length, caudal fin length and average length of paired pectoral fins were highly significantly different (p < 0.01) while anal fin length was significant different (p < 0.05) among the experimental sites. In case of C. mrigala, the body weight was non-significantly different (p > 0.05) while all other parameters were highly significantly different (p < 0.01) except the dorsal fin length which was significantly different (p < 0.05) among the study sites.
2. The results of the Pearson correlation of morphometric parameters showed that body weight of L. rohita developed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters, the fork length of the said species showed a positive and highly significant (p< 0.0001) correlation with all the parameters except with the caudal fin length where the correlation was also positive but non-significant (p = 161). In case of total length of the fish body, the correlation was highly significant (p< 0.0001) and positive with all the parameters of study.The length of the dorsal fin showed highly significant (p< 0.0001) and positive correlation with all the remaining morphometric parameters under study. The caudal fin length of L. rohita showed a positive and highly significant (p< 0.0001) correlation with all the other parameters except the fork length where the correlation was positive but non-significant (p = 161). The correlation of the anal fin length of the fish body showed a highly significant (p< 0.0001) and positive correlation trends. The average length of the paired pectoral fins showed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters of study, the correlation of paired pelvic fins average length showed positive and highly significant (p< 0.0001) correlation with other parameters.
3. The body weight of C. mrigala developed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters. The fork length of the said species showed a positive and highly significant (p< 0.0001) correlation with body weight, total length and dorsal fin length while this correlation was positive but non significant with the caudal fin length (p = 0.228), anal fin length (p = 0.168), average length of paired pectoral fins (p = 0.031) and average length of the paired pelvic fins (p = 0.106). In case of total length of the fish body, the correlation was highly significant (p< 0.0001) and positive with all the parameters of study. The length of the dorsal fin showed highly significant (p< 0.0001) and positive correlation with all the remaining morphometric parameters under study. The caudal fin length of C. mrigala showed a positive and highly significant (p< 0.0001) correlation with all the other parameters except the fork length where the correlation was positive but non-significant (p = 0.228).The correlation of the anal fin length of the fish body showed a highly significant (p< 0.0001) and positive correlation trends with all the parameters except the fork length where the correlation was positive but non-significant (p = 0.168). The average length of the paired pectoral fins showed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters of study except the fork length where the correlation was positive but non-significant (p = 0.031). InC. mrigala, the correlation of paired pelvic fins average length showed positive and highly significant (p< 0.0001) correlation with other parameters except the fork length where the correlation was positive but non-significant (p = 0.106).
4. Dendrogram generated on the basis of morphometric parameters of study dividedL. rohita genotype in to five major clusters or classes with 19.24% for within class variation while 80.76% for the between class differences. While the dendrogram developed for C. mrigala divided the genotypes in to four major clusters or classes with 27.28% for within class variation while 72.72% for the between class differences.
5. The results obtained from the PCA for morphometric parameters of L. rohitaand C. mrigalaindicated clearly that the increase in the number of factors or components was correlated with the decrease in eigenvalues. The values showed that its trend reached its maximum at level of second factor. In the same way according to the Kaiser (1958) criterion based upon the eigenvalues greater than one, first two main factors accounted for 80.273% of cumulative variability for L. rohita and 82.558% for C.mrigala. The PCA grouped the tested variables or parameters of the L. rohita,the first group amongst the major two groups accounted for 64.245% of the cumulative variability while the second from these accounted for 16.028% of the cumulative variability. The PCA grouped C. mrigala,also into two groups, the first group amongst the major two groups accounted for 59.323% of the cumulative variability while the second from these accounted for 23.235% of the cumulative variability.
6. The physico-chemical parameters of the water samples of all study sites were analyzed for correlation among them. The results were as follows; the correlation of the pH with water temperature (r= 0.107) and dissolved oxygen (r = 0.905) was positively non-significant while the correlation with electrical conductivity (r = -0.798), salinity (r= -0.888), total dissolved solids (r = -0.857), total alkalinity (r = -0.736) and total hardness (r = -0.499) was negatively non-significant. The correlation of the dissolved oxygen with water temperature (r= 0.313) was positively non-significant while the correlation with electrical conductivity (r = -0.669), salinity (r= -0.828), total dissolved solids (r = -0.809), total alkalinity (r = -0.930) and total hardness (r = -0.300) was negative but also non-significant as like with the water temperature. The electrical conductivity was positively correlated with all the physic-chemical parameters as with water temperature (r= 0.482), salinity (r= 0.925), total dissolved solids (r = 0.889), total alkalinity (r = 0.452) and total hardness (r = 0.906) and this correlation was non significant.The salinity amongst the water parameters was correlated positively with water temperature (r = 193), total alkalinity (r = 0.717) and total hardness (r = 0.734) and it was non-significant but with total dissolved solids (r = 0.994) the correlation was also positive but highly significant (P < 0.001). The total dissolved solids values observed from the study sites were positively correlated with water temperature (r = 0.172), total alkalinity (r = 0.734) and total harness (r = 0.657) and this correlation was non-significant. The correlation between the total alkalinity and total hardness was also positive and non-significant (r = 0.048).
RAPAD Data
1. In case of L. rohita, OPB-1 polymorphism remained as 16.67%, OPB-3 polymorphism remained as 40.00%, OPB-4, polymorphism remained as 16.67%, OPB-5 polymorphism remained as 20.00%, OPB-7 polymorphism was 28.57%, OPB-8 polymorphism was 20.00%, OPB-9 polymorphism was 25.00%, OPB-10 polymorphism was 28.57%, OPC-19 polymorphism was 14.29% and OPD-4 showed 50.00% polymorphism in amplification. In case of C. mrigala, OPB-1 polymorphism remained as 16.67%, OPB-3 polymorphism remained as 16.67%, OPB-4 polymorphism remained as 25.00%, OPB-5 polymorphism remained as 14.29%, OPB-7 polymorphism was 14.29%, OPB-8 polymorphism was 20.00%, OPB-9 polymorphism was 20.00%, OPB-10 polymorphism was 20.00%, OPC-19 polymorphism was 28.57% and OPD-4 polymorphism remained as 33.33% in amplification.
2. The dendrogram generated by UPGMA of RAPAD data of L. rohita by the randomly selected individuals with high scorable bands of the five populations grouped themselves in the first class/cluster while a single sample designated as Indus2 from the population from River Indus collected from Taunsa Barrage represents the second class/cluster and in same way only single individual designated as Ravi2 collected from River Ravi from the Baloki Barrage represents the third class. The dendrogram generated by UPGMA of RAPAD data of C. mrigala by the randomly selected individuals of the five populations grouped themselves in the first class/cluster and two samples designated as Indus2 and Qad2 from the populations from River Indus collected from Taunsa Barrage and River Chenab from Qadirabad Barrage represents the second class/cluster while one individual from the Trimu Barrage at the junction of Jhelum and Chenab Rivers designated as Trimu2 represents the third class and in the same way only single individual designated as Ravi2 collected from River Ravi from the Baloki Barrage represents the third class.
3. The PCA resultsfor L. rohitait can be assumedthat PCA grouped the tested variables or parameters of the fish RAPAD amplification data into two main components which all together accounted for 58.177% of the cumulative variation among the factors. The first group (F1) amongst the major two groups accounted for 33.327% of the cumulative variability while the second (F2) from these accounted for 24.850% of the cumulative variability. These results were also confirmed after the varimax rotation. By the PCA resultsfor C. mrigalawe can assume after observing the results that the PCA grouped the tested variables or parameters of the fish RAPAD amplification data into two main components which all together accounted for 70.866% of the cumulative variation among the factors. The first group (F1) amongst the major two groups accounted for 51.115% of the cumulative variability while the second (F2) from these accounted for 19.751% of the cumulative variability.
This study in this way has provided the genetic information of the present fish species and how evolutionary processes are affecting the fish fauna. So this study along with the strengthening of the academic research area has also proven an applied research which will help the breeders to the chose most fit candidates for the breeding program in the Pakistan.
Availability: Items available for loan: UVAS Library [Call number: 1504,T] (1).
2.
Intraspecific Genetic Variations Amongst The Populations Of Labeo Rohita
by Syed shafat hussain | Dr. Fayyaz rasool | Dr. Noor khan | Mr. Hamid mustafa.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1782,T] (1).
3.
Mapping The Genetic Diversity Of The Populations Of Cirrhinus Mrigala Through Mitochondrial Atpase 6/8 Genes From Indus Riverine System Of Pakistan
by Noor Muhammad (2014-VA-543) | Dr. Fayyaz Rasool | Mrs. Shakeela Parveen | Dr. Muhammad Yasir Zahoor.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: This study was mapping the genetic diversity of Cirrhinus mrigala by using molecular markers for the ATPase 6/8 gene region. Samples of the target fish specie were collected from the different sites at the Indus riverine system of the Pakistan i.e Chenab (Head Qadirabad), Jhelum(Head Rasul), Ravi(Head Balloki) and Indus (Head Taunsa). Morphommetric parameters viz., Body weight, Girth, Total Length, Fork Length,Head length, Lengths of Caudal, Pectoral, Ventral Fins were recorded manually using measuring scale. After recording morphometric parameters blood from fish samples was drawn from caudal fin of the Fish, collected in Vautainer and immediately transferred into the Lab of IBBT where it is preserved in -80C for future use in DNA extraction. Phenol Chloroform Method of Genomic DNA extraction was used to extract whole blood DNA. The extracted DNA of the samples was run on 0.8% agarose gel by using electrophoresis of the DNA. Nanodrop was used to find out the concentration of DNA after that DNA was diluted according to the Need with deionized water. The PCR of ATPase 6/8 primers were used to amplify the target gene. The PCR product was confirm on 1.2% agarose gel. These gels were visualized in UV light and photographs were taken by Gel Documentation System (BIO-RAD, Gel-DOC EZ imager). After the visualizing of quality of band confirmation the samples were sent to be sequencing. The Analysis of Variance (ANOVA) for the different morphometric parameters was done by XLSTAT 2012 version 1.02. ANOVA results showed that all the morphomteric parameters were highly significant (p<0.01) within as well as between the groups. Pearson correlation analysis off overall population show that the body weight is positively correlated with the Fork Length, Total Length, Ventral Fin length, caudal Fin Length, Girth, with significant difference. Fork length is positively correlated with Total length, Ventral
Fin Length, Caudal Fin length, Pectoral Fin Length, with significant difference among other parameters. Total Length is positively correlated with Girth, Pectoral Fin Length, Ventral Fin Length, Caudal Fin Length, Girth, and Weight with significant difference among them. NJ methods suggested that existed relationship among different species located at different geographical locations. They all have a common ancestry. A strong relationship exists between fish species collected from different locations at Indus riverine system of Pakistan.
Availability: Items available for loan: UVAS Library [Call number: 2479-T] (1).
4.
Amphibians And Reptiles Of Kalabagh, District Mianwali, Punjab
by Muhammad Farooq (2014-VA-554) | Dr. Waseem Ahmad Khan | Dr. Shahzad Ali | Dr. Fayyaz Rasool.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The present study was conducted in the Kalabagh game reserve (KGR) from July 2015 to April 2016. The surveys were conducted with the help of managing and supporting staff from Kalabagh wildlife Foundation in order to know the diversity and various threats to amphibians and reptiles in the study area.
A total 23 species belonging to three orders, 16 families and 23 genera were recorded including six amphibians and 17 reptiles. Amphibians included one toad and five frogs whereas; reptiles included one turtle, ten lizard and six snake species. Three species out of the six recorded snakes are venomous rests all the amphibian and reptiles are non-venomous. All the recorded species were captured directly.
Amphibians were searched actively in their potential breeding areas and resting places like ponds, water channels and along streams during day and night. Simple hand nets or scoop nets were used for amphibian collection; however, calling males are best caught by hands. Hand picking through bare hands or with the help of long forceps or snake clutch has always been the most efficient way of collecting different reptiles. . Snakes were mostly collected using snake catcher and every snake being collected was considered as venomnous in order to avoid any mishap. Road-Killed snakes were also collected.
Following the specific criteria , two species were evaluated as Abundant, eight species as Common, and five species as Fair, one species as Rare and seven species as Occasional.
Out of the recorded 23 species, two reptiles were found among endemic species to Pakistan including one lizard; Salt range ground gecko and one snake; slender blind snake.
Six of the recorded 23 species are CITES Species with one species (Varanus bengalensis) is enlisted in Appendix I, while five species (Hoplobatrachus tigerinus, Lissemcieys punctata, Eryx johnii, Uromastyx hardwickii and Naja naja,) are enlisted in Appendix II of the CITES category 2016. The only one species Varanus bengalensis is having decreasing IUCN population trend.
There are some potential threats to herps in the study area. Some snakes that get trapped are killed by the local residents. The Spiny tailed lizard is also captured by some hunters for sale to local Hakeems to extract oil. Black cobra is captured by snake charmers to entertain people and for extraction of venom. The evidences reveal that the populations of various amphibians and reptiles including snakes in the study area are not facing any decline rather they are stable except spiny tailed lizard and Bengal monitor. Most of the recorded herps from the study area also occupy a vast distribution in the country. No crocodilians and tortoises are found in the study area. The existing herps include; frogs, toads, freshwater turtles, lizards and snakes.
Availability: Items available for loan: UVAS Library [Call number: 2478-T] (1).
5.
Assessment Of Metalic Loads In Water Sediments And Fish Sampled From River Ravi, Pakistan With Study On Their Mitigation Methods
by Muhammad Shakir (2007-VA-320) | Dr. Hamda Azmat | Dr. Fayyaz Rasool | Dr. Arshad Javid.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Present study on the detection of heavy metals in water, sediments and fish, Cirrhinus mrigala
samples was conducted at river Ravi along with its three different sights (Ravi Siphon, Saggian
Bridge and Balloki Headworks). The stations were further divided into two sub-stations viz. (i)
Upstream and (ii) Downstream. Water and sediment samples were collected from three different
points from the sub-stations.
Water, sediment and fish samples were collected on monthly basis from November, 2015
through February, 2016. Average chromium concentrations in water samples from Baloki
Headworks during November, December, January and February were 0.33±0.17 mgL-1,
0.18±0.03 mgL-1, 0.17±0.06 mgL-1 and 0.17±0.06 mgL-1, respectively. Similarly, average
chromium concentrations from November, 2015 through February, 2016 from Saggian Bridge
was recorded 0.27±0.13 mgL-1, 0.18±0.04 mgL-1, 0.22±0.06 mgL-1 and 0.18±0.10 mgL-1,
respectively. From Shahdera bridge, the chromium concentrations of 0.28±0.12 mgL-1,
0.22±0.04 mgL-1, 0.23±0.06 mgL-1 and 0.16±0.07 mgL-1 were recorded during November,
December, January and February, respectively. Over all mean Cr concentrations in water
samples from Balloki Headworks, Saggian Bridge and Shahdera Bridge from November, 2015
through February, 2016 were recorded as 0.30±0.14 mgL-1, 0.19±0.04 mgL-1, 0.21±0.06 mgL-1
and 0.17±0.08 mgL-1, respectively.
During monthly surveys, average Ni concentrations in water samples collected from Baloki
Headworks were recorded 0.24±0.11 mgL-1, 0.27±0.15 mgL-1, 0.22±0.13 mgL-1 and 0.21±0.10
mgL-1 during November, December, 2015 and January and February, 2016, respectively. Water
samples from Saggian Bridge had Ni concentrations of 0.23±0.14 mgL-1 during November,
2015, 0.25±0.16 mgL-1 during December, 2015, 0.23±0.18 mgL-1 January, 2016 and 0.20±0.16
SUMMARY
36
mgL-1 during February, 2016. Water samples from Shahdera Bridge contained Ni concentrations
of 0.27±0.14 mgL-1, 0.28±0.17 mgL-1, 0.26±0.16 mgL-1 and 0.20±0.16 mgL-1, respectively from
November, 2015 through February, 2016, respectively.
Average Pb concentrations in water samples from Baloki Headworks from November, 2015
through February, 2016 were recorded 0.14±0.12 mgL-1, 0.08±0.04 mgL-1, 0.06±0.05 mgL-1 and
0.16±0.06 mgL-1, respectively. Similarly, average Pb concentrations from Saggian Bridge
sampling stations were recorded 0.23±0.14 mgL-1, 0.25±0.16 mgL-1, 0.23±0.18 mgL-1 and
0.20±0.16 mgL-1, during November, December, January and February, respectively. The Pb
concentrations, from Shahdera Bridge water samples from November, December, January and
February were recorded 0.27±0.14 mgL-1, 0.28±0.17 mgL-1, 0.26±0.16 mgL-1 and 0.20±0.16
mgL-1, respectively.
Average Cr concentrations during the study period in sediment samples from Balloki
Headworks, Saggian Bridge and the Shahdara Bridge were recorded 0.27 mgL-1, 0.22 mgL-1 and
0.29 mgL-1, respectively. Similarly, average Ni concentrations from Balloki Headworks, Saggian
Bridge and Shahdara Bridge were recorded 0.13 mgL-1, 0.132 mgL-1 and the 0.13 mgL-1,
respectively. Average Pb concentrations from Balloki Headworks were recorded 0.20 mgL-1,
from Saggian Bridge 0.22 mgL-1 while it was noted 0.19 mgL-1 from Shahdara Bridge.
During the month of November, 2015 higher Cr concentrations were recorded from muscle as
compare to the other body organs. Similarly, liver showed maximum Pb accumulation while
higher Ni concentrations were observed from liver and muscles. During December, 2015,
maximum Cr concentration was recorded in gills and skin while higher Pb concentrations were
recorded from the liver. Similarly, Ni concentration was higher in liver as compared to the other
body organs. During January, 2016 maximum Cr concentration was recorded from liver while Pb
SUMMARY
37
and Ni concentrations were recorded maximum from skin of the Cirrhinus mrigala. During
February, higher Cr, Pb and Ni concentrations were recorded from liver as compare to all the
other fish organs.
Statistically significant variations in metal ion absorption were recorded between different
sampling stations. Significantly higher (p<0.05) Ni concentrations were absorbed by alkali
treated banana peels from the water samples collected from all the three sampling stations.
Similarly, higher Cr concentrations were absorbed through the alkali treated orange peels from
Baloki Headworks.
Statistically significant (p<0.05) variations in absorption of heavy metals from water samples
was recorded between the sampling stations. Among all the three heavy metals, significantly
lower Pb concentrations were absorbed through acid treated banana and orange peels from all the
three sampling stations. Availability: Items available for loan: UVAS Library [Call number: 2487-T] (1).
