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The number of pollutants released into freshwater and marine environments has increased due to the widespread use of nanoparticles. Nickel oxide nanoparticles (NiO-NPs) were tested for genotoxicity in fish fingerlings of the species Ctenopharyngodon idella. For 7, 14, and 21 days, fingerlings were exposed to NiO-NPs with each increasing concentrations of 2.25 mg/L, 4.50 mg/L, and 6.75 mg/L, respectively. The micronuclei assay and comet assay were used to evaluate the DNA damage. The experiment revealed that with the increase in nanoparticle concentration and exposure duration, the level of DNA damage also increased. The experiment resulted to be time and dose dependent, and the damage was found as follows: 6.75 mg/L > 4.50 mg/L > 2.25 mg/L against each exposure period. In terms of comet assay, the results showed that after 7 days, the level of DNA damage in all the concentrations was highly significant (P < 0.001). Increased DNA damage was calculated at the higher administered dose of 6.75 mg/L for 21 days of exposition, followed by 14 and 7 days, respectively. The second high toxic effect was observed in the fish blood at the exposure concentration of 4.50 mg/L for 21 days, followed by 14 and 7 days, respectively. The micronuclei induction in the nanoparticle's administered blood could be detected only for a 7-day exposition period. Whereas for the exposed duration of 14 and 21 days, the entire red blood cells of the grass carp were completely destroyed demonstrating the ability of the nanoparticles to cause anomalies in aquatic life., Competing Interests: Declarations. Ethical Approval: Ethical approval for the study was taken from the Ethical Committee, University of Peshawar. Competing Interests: The authors declare no conflict of interest., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Grass carp (Ctenopharyngodon idella) was exposed to nanoparticles of different concentrations, i.e., copper oxide nanoparticles (CuO-NPs), silver nanoparticles (Ag-NPs), and green synthetic gold nanoparticles (Au-NPs).The administered doses of the three concentration groups were 20mg L -1 , 30 mg L -1 , and 40mg L -1 each for a period of 14 and 28 days, respectively. The DNA damage in the erythrocytes of grass carp was detected through a comet assay technique. The values of total comet score (TCS) were noted for the exposed concentrations with a significant increasing trend (p < 0.05) and ordered as CuO-NPs > Ag-NPs > Au-NPs. The highest TCS value of the exposed erythrocytes was recorded for CuO-NPs at 40 mg L -1 after 14 days of exposition period. Comparatively, TCS values of erythrocytes exposed to green synthetic Au-NPs at all the concentrations and exposed time were less as compared to the Ag-NPs and CuO-NPs. The study confirmed the adverse effects of nanoparticles on the genetic material of fish cells., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Intellectual disability (ID) is a large group of neurodevelopmental disorders characterized by a congenital limitation in intellectual functioning (reasoning, learning, and problem solving), adaptive behavior (conceptual, social, and practical skills), originated at birth and manifested before the age of 18. By whole exome sequencing of five consanguineous Pakistani families presenting hallmark features of ID, global developmental delay, aggressive and self-injurious behaviors, microcephaly, febrile seizures and facial dysmorphic features, we identified three novel homozygous missense variants (NM&#95;024298.5: c.588G > T; p.Trp196Cys, c.736 T > C; p.Tyr246His and c.524A > C; p. Asp175Ala) and one rare homozygous in-frame deletion variant (c.758&#95;778del;p.Glu253&#95;Ala259del) in membrane-bound O-acyltransferase family member 7 (MBOAT7) gene previously associated with autosomal recessive neurodevelopmental disorder. The segregation of the variants was validated by Sanger sequencing in all family members. In silico homology modeling of wild-type and mutated proteins revealed substantial changes in the structure of both proteins, indicating a possible effect on function. The identification and validation of new pathogenic MBOAT7 variants in five cases of autosomal recessive ID further highlight the importance of this genes in proper brain function and development., (© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Oculocutaneous albinism (OCA) is a group of skin depigmentation disorders. Clinical presentation of OCA includes defects in melanocyte differentiation, melanin biosynthesis, and melanosome maturation and transport., Objectives: A molecular diagnostics study of families presenting oculocutaneous albinism., Methods: In this study, 17 consanguineous OCA families consisting of 93 patients were investigated. Whole Exome Sequencing (WES) of the index patient in each family were performed. Short listed variants of WES were Sanger validated for Mendelian segregation in obligate carriers and other available family members. Variant prioritization and pathogenicity were classified as per the criteria of American College Medical Genetics and Genomics (ACMG). Comparative computational modelling was performed to predict the potential damaging effect of the altered proteins., Results: 15 pathogenic variations: c.132 T > A, c.346C > T, c.488C > G, c.1037G > A in TYR, c.1211C > T, c.1441G > A, c.1706&#95;1707insT, c.2020C > G, c.2402G > C, c.2430del, in OCA2, c.1067G > A in TYRP1 and c.451C > T, c.515G > T, c.766C > T, c.917G > A in MC1R genes were identified. Three variants in OCA2 gene were characterized: c.1706&#95;1707insT, c.2430del, and c.2402G > C, all of which were not reported before in OCA families., Conclusion: A few studies focusing on mutation screening of OCA patients have been reported before; however, this study has uniquely presents the Pakhtun ethnic population residing on the North-Western boarder. It explains that TYR, OCA2, TYRP1, and MC1R variations lead to non-syndromic OCA phenotype The overlapping phenotypes of OCA can precisely be diagnosed for its molecular pathogenicity using WES. This study recommends WES as a first-line molecular diagnostic tool, and provides a basis for developing customized genetic tests i.e. pre-marital screening to reduce the disease burden in the future generations., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Vehicle-wash wastewater (VWW) contains high levels of various petrochemicals such as polycyclic aromatic hydrocarbons (PAHs), a carcinogenic category of organic substances. However, the genotoxic effects of PAHs present in VWW remain largely unknown. We explored the genotoxic effects of PAHs present in VWW on fish grass carp (Ctenopharyngodon idella) and freshwater mussels (Anodonta cygnea). Fish and freshwater mussels were divided into control and exposed groups, the prior groups were treated at weekly intervals with clean water, and the latter with Σ16PAHs contaminated VWW for up to four weeks. The samples of blood from fish and haemolymph from freshwater mussels were collected and analyzed using the comet assay technique. Results exhibited that in control fish and freshwater mussel groups the genotoxicity decreased with every week passing following the order of W1 > W2 > W3 > W4, ranging from 8.33 ± 3.06 to 25.3 ± 4.62 and from 46.0 ± 6.93 to 7.67 ± 3.79, respectively. The exposed fish and freshwater mussel groups indicated an increase in genotoxicity with increasing week intervals with an order of W4 > W3 > W2 > W1, ranging from 55.7 ± 11.9 to 128.3 ± 10.0 and from 112.7 ± 8.50 to 183.3 ± 10.1, respectively. The genotoxic effect of Σ16PAHs on fish was comparatively lower than on freshwater mussels. This study elucidates that VWW is highly genotoxic and should be treated before discharging into aquatic ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Based on newly provided morphological and previously published genetic data, we describe two new distinctive sympatric lacertid lizards of the genus Eremias (subgenus Aspidorhinus) from the arid mountains of northwestern Balochistan Province, Pakistan. The new species, Eremias killasaifullahi sp. nov. and Eremias rafiqi sp. nov. are distinguished from all other species of the subgenus Aspidorhinus (E. afghanistanica, E. fahimii, E. isfahanica, E. kopetdaghica, E. lalezharica, E. montana, E. nikolskii, E. papenfussi, E. persica, E. regeli, E. roborowskii, E. strauchi, E. suphani, and E. velox) by unique morphological characters and genetic differentiation. In the molecular phylogeny of Aspidorhinus, both new taxa are well-supported lineages differentiated from other species of this subgenus by uncorrected p distances from 8.5% to 21.6%, respectively. Both new species belong to E. persica complex where E. rafiqi sp. nov. is partly similar in dorsal color pattern to E. persica but can be distinguished from this species by unique meristic and morphometric characters. Eremias rafiqi sp. nov. is found in Pakistan, Afghanistan, and eastern Iran. Eremias killasaifullahi sp. nov. can be differentiated from E. persica by its distant distribution, dorsal color and pattern, smaller size, and less number of gulars and ventral scales. Eremias killasaifullahi sp. nov. is only known from the type locality and represents the local microendemism, along with other endemic species of reptiles reported from this part of Pakistan. We, however, expect that E. killasaifullahi sp. nov. could have a broader range in northwestern Pakistan and southeastern Afghanistan, which should be an object of following investigations. Our data show that remote areas between Hindu Kush Mountains and Indus River need attention as they most probably represent possible sources of genetic and species diversity in the region.

The current study aimed to investigate the relationship between polymorphisms in detoxifying (GSTM1, GSTT1, and GSTP1) genes and their association with colorectal cancer (CRC) in tobacco addicts of Pashtun ethnicity. Polymorphisms in the selected genes were genotyped in a case-control study consisting of 100 histologically confirmed male CRC patients and 100 birth-year and gender-matched healthy controls using the PCR-RFLP method. The GSTM1 null, and GSTT1 null genotypes were significantly contributed to the risk of CRC in the cases (OR= 3.131, 95% CI: 1.451-6.758, P = 0.004, and OR= 3.541, 95% CI: 1.716-7.306, P = 0.001, respectively), whereas the association observed for GSTP1 Val/Val (1.139, 95% CI: 0.356-3.644, P = 0.826) did not show statistical significance. The combined GSTM1 null and GSTT1 null showed a 41-fold increased risk (95% CI: 4.945-351.950, P = 0.001), while, the combined GSTM1 null and GSTP1 Ile/Val or Val/Val variant genotypes exhibited about 3-fold (95% CI: 1.196-7.414, P = 0.019) increased risk to CRC. Similarly, the combined GSTT1 null and GSTP1 Ile/Val or Val/Val variant genotypes showed about a 3-fold (95% CI: 1.285-8.101, P = 0.013) increased risk of CRC. In the combination of three GST genotypes, the GSTM1 null, GSTT1 null, and GSTP1 Ile/Val or Val/Val variant genotypes demonstrated a more than a 22-fold (95% CI: 2.441-212.106, P = 0.006) increased risk of CRC. Our findings suggest that GSTM1 and GSTT1 polymorphism and its combination with GSTP1 may be associated with CRC susceptibility in the Naswar addicted Pashtun population of Khyber Pakhtunkhwa, Pakistan.

Grass carps were exposed to the established lethal concentration (LC) values of copper (Cu), chromium (Cr), and lead (Pb) each for the exposed period of 24, 48, 72, and 96 h respectively. Concentrations of these metals were determined in the brain, liver, muscles, gills, kidneys, and intestinal tissues of exposed and control fish through the atomic absorption spectrophotometer after the wet digestion process. The metals accumulation inside these tissues confirmed the absorption of metals from media into the tissues of the model organism. The accumulated concentration in fish tissues was confirmed to be concentration-dependent with significant (p < 0.05) elevated mean values seen for the lead followed by chromium and copper as compared with the mean concentration values of their respective control group. Levels of metals were found above the permissible standards suggested by the regulatory authorities in the fish's body. Histological sections of the same targeted organs exposed to the three exposure concentration groups were studied and compared with the sections of the healthy group. The histopathological lesions were scored to rank the deleterious effects of metals. The histopathological changes were recorded in concentration and progressive time-related series where gills had the greatest number of scored lesions followed by the kidneys and intestines, muscles, brain, and finally the liver as the least affected organ. Moreover, the organs were not affected uniformly by the metals; in fact, every studied organ has given mild to severe responses towards the toxic metals where lead had proven to cause more severe lesions as compared with copper and chromium. The histological lesions recorded mostly were thus concentration-dependent as revealed in the bioaccumulation of these metals with the effects ranked as lead > chromium > copper with a few exceptions. The findings can be used as a benchmark for the evaluation of the fate and effects of the toxic metals in the expanded aquaculture production of grass carp nationwide. Further investigations with respect to other potentially toxic metals like arsenic, mercury, and cadmium could address the problem towards additional studies.

The genus Heremites Gray, 1845 is endemic to the Western Palearctic region, containing morphologically similar species with a not well resolved taxonomy. The genus has a broad distribution from North Africa to Central Asia, with the only known record from northeastern Afghanistan. Three species are currently recognized in the genus with one, H. septemtaeniatus (Reuss, 1834), representing populations at the eastern edge of the genus range. During extensive fieldwork, we discovered H. septemtaeniatus from northwestern Pakistan and provisionally suggest that this population could be morphologically defined as H. septemtaeniatus transcaucasicus (Chernov, 1926). This important contribution to the knowledge regarding the family Scincidae in Pakistan, however, needs further investigation using an integrative approach., (Rafaqat Masroor, Muhammad Idrees, Muhammad Khisroon, Qaisar Jamal, Daniel Jablonski.)

Welding is used throughout the world in refineries, thermal power plants, chemical facilities, and pressurized containers, and the welders are exposed to toxic heavy metals, electromagnetic fields, polycyclic aromatic hydrocarbon, and ultraviolet radiations. In the present study, 59 welders and an equal number of control subjects were assessed for DNA damage in the lymphocytes using the comet assay. Heavy metals such as lead (Pb), iron (Fe), nickel (Ni), chromium (Cr), manganese (Mn), and cadmium (Cd) levels in the scalp hair of the subjects were evaluated by using atomic absorption spectroscopy (AAS). The results of the current study showed that DNA damage in the lymphocytes of welders (121.8 ± 10.7) was significantly higher as compared with controls (56.5 ± 17.6) (P < 0.001). Besides, the levels of Pb, Fe, Ni, Cr, Mn, and Cd were remarkably higher in the scalp hair of workers as compared with the control group (P < 0.001). Regression analysis showed a prominent association between the heavy metals and total comet score (TCS) in the exposed subjects. Age and duration of occupational exposure had significant effects (P < 0.05) on TCS values. Our results concluded that occupational exposure to welding fumes may cause DNA damage and can lead to important health hazards in the workers.

Endosulfan is an organochlorine pesticide, which is commonly used throughout the world. It accumulates in the environment and may cause significant damage to the ecosystems, particularly to the aquatic environments. The present study was conducted to evaluate the genotoxic effect of endosulfan on the grass carp (Ctenopharyngodon idella) blood. The fish were exposed to three different concentrations, 0.75 ppb/day, 1.0 ppb/day, and 1.5ppb/day of endosulfan for 7, 14, 21, and 28 days. The study was a randomized control trial and the control group was not exposed to endosulfan. The results showed that after 7 days, the level of DNA damage in all the concentrations was significant (P < 0.05), while after 14, 21, and 28 days' trials, highly significant (P < 0.000) level of DNA damage was observed. Hence, time- and dose-dependent DNA damage was observed in fish DNA by comet assay. It is concluded from our results that with the increase in endosulfan concentration and exposure duration, the level of DNA damage also increased. As the current study showed the severe genotoxic effect of endosulfan in Ctenopharyngodon idella, therefore, the imprudent and indiscriminate use of endosulfan should be controlled and monitored by the concerned government authorities.

The primary aim of the current study was to recognize the biomarker approach as the finest tool to identify the geno-toxicological effects of copper, chromium, and lead inside the blood of grass carp using micronucleus test and comet assay. The induced micronuclei frequency in response to the administered concentrations of LC50 metals was discovered in the erythrocytes of metal-exposed fish at four-time intervals. The genotoxic effect of these metals with respect to the formation of micronuclei was ranked as chromium > lead > copper. Percentages of other cellular and nuclear abnormalities were also determined in the exposed blood films. Equally, the genotoxic studies in terms of comet assay in fish blood revealed significant deviations p < 0.05 against each of the studied metal at their respective time interval as compared with the healthy fish group. However, induced frequency of micronuclei and the calculated DNA damage were not found to be duration dependent. Consequently, copper, chromium, and lead have been explored as cytotoxic elements that can be responsible for inducing genotoxic effects in fish existing aquatic habitats.

The current study determines the possible antitumor and immunomodulatory effects of thymosin against the in vivo and in vitro growth of tumor-derived cell line in mice. Peritoneal phagocytes count, Ehrlich ascites tumor (EAT) cells, T- lymphocytes, and B- lymphocytes activities were determined. In addition, serum level of interleukin 2 (IL-2) and liver functions were measured. In animal testing, thymosin at doses of 0.50 and 1mg activated the phagocytic function of macrophages, as well as T- and B- cell function. Thymosin caused a marked shortage in the proliferation of EAT cells in the peritoneal fluid with dose 0.50g as compared with that of the corresponding control group. Furthermore, treatment with thymosin caused effectively elevate in serum level of IL-2, on the contrary reduce in serum levels of ALT, AST and total proteins. The size of solid Ehrlich tumor was significantly decreased, as measured morphologically with the doses 0.50 and 1 mg (P<0.01). These results confirmed that many biological activities attributed to thymosin and is as an adjuvant for immune enhancement.

Objectives: The objective of this study was to assess the influence of antioxidant gene GSTM1 and GSTT1 on DNA damage in personnel occupationally exposed to volatile anaesthetics (VA)., Methods: The study groups were composed of 50 exposed subjects (anaesthesia workers) and 49 controls. Blood samples were collected from both subjects. DNA damage was analysed through the comet assay technique. Biomarker genes GSTM1 and GSTT1 were inspected through PCR technique for polymorphism., Results: The comet assay technique showed that the Total Comet Score (TCS) in exposed subjects was significantly higher (p=0.0001) than the control. Age and smoking had significant effects on TCS in the study groups (p<0.05). Duration of occupational exposure had significant positive correlation (r=0.755, p<0.001) with DNA damage. The null polymorphism in GSTM1 and GSTT1 gene showed a significant effect (p<0.001 and p<0.000) on the DNA damage., Conclusions: The polymorphism in GSTM1 and GSTT1 gene significantly damage DNA in personnel occupationally exposed to VA., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

Objectives: The study was performed to know about the extent of occupational DNA damage in photocopiers' workers., Methods: Blood samples were collected from 136 exposed group and 74 control group. Comet assay was performed to assess the DNA damage caused by emissions from photocopiers., Results: The results indicated that there was a significant increase (P < 0.05) in DNA damage in persons working in photocopiers (122.1 ± 60.7) than the controls (56.6 ± 17.2). Duration of occupational exposure had positive correlation (r = 0.793, P < 0.001) with DNA damage. Age had significant effects on the total comet score (TCS) of the exposed group as compared to the control group (P < 0.05)., Conclusions: In conclusion, these findings indicate significant genotoxicity in photocopiers' workers.

Purpose: To evaluate the genotoxic effects of gold jewellery fumes and its association with GSTM1 and GSTT1 genetic polymorphisms., Materials and Methods: We examined 94 subjects including 54 gold jewellery workers and 40 controls. The DNA damage was evaluated by alkaline comet assay and genotyping by PCR., Results: The mean total comet score (TCS) in gold jewellery workers was significantly higher as compared to the control subjects (128.0 ± 60.6 versus 47.7 ± 21.4; p  = 0.0001). Duration of occupational exposure had positive correlation ( r  = 0.453, p  < 0.01) with DNA damage. Age and tobacco use had significant effects on the TCS of the exposed group as compared to the control group ( p  < 0.05). The frequency of the GSTM1-null genotype in the exposed group was significant ( p  = 0.004) as compared to the control group. No significant association ( p  > 0.05) between the GSTM1 and GSTT1 genotypes and DNA damage was found., Conclusions: Our results suggest that there is increased DNA damage in gold jewellery workers due to their occupational surroundings. Hence there is a strong need to educate the workers about the adverse health effects of potentially hazardous chemicals and highlight the importance of using protective measures.

Hematological variances were seen in Ctenopharyngodon idella when exposed to (LC50 and LC85) copper, chromium, and lead concentration groups. The metal-exposed blood samples after collection were analyzed for comparative analysis of hematological alterations in the reference and treated groups at the different exposure hours. A significant reduction (p < 0.05) in hemoglobin, RBC, hematocrit, total leucocyte, and lymphocyte counts was observed with the increase in metal concentrations, whereas a significant incline (p < 0.05) in granulocyte and monocyte counts was observed with the administered doses of selected toxic metals as compared with the values obtained from the reference group. More pronounced changes in hematological indices both for the red and white blood cells were noticed at the highest dose of LC85-administered metal groups. The induced hematological changes were independent of the exposed duration pattern. The present investigation has significantly contributed in knowledge assessment of metal toxicity based on inclusive evaluation of dose-response relationships.

In the current study, chlorpyrifos was used as a test chemical to evaluate its possible toxicological effect on birds. A total of 45 adult male Japanese quails were divided into five groups (A to E). Each group, containing 9 birds was further divided into 3 sub-groups (containing 3 birds each). Group A served as control, while all other groups and sub-groups were exposed to selected pesticide for different trial periods. Chlorpyrifos sub-lethal doses were orally administered daily at the rate of 3, 6, 9, and 12 mg/kg body weight per day to group B, C, D, and E, respectively. Birds were kept under observation for behavioral changes throughout the trial periods. Clinical signs, histological alterations, genotoxicity, and blood biochemical alterations were recorded after each 15-day trial. Mild to moderate clinical signs like staggering gait, tremors, diarrhea, dullness, less frequency of crowing, and decrease foam production were observed in group D and E throughout the study. The changes in the body weight gain and blood biochemical parameters among different groups at a given trial period were insignificant. The appearance of micronuclei in group E birds was more significant, indicating that nucleus damage was dose-dependent while to lesser extent duration-dependent. The comet assay showed significant dose- and duration-dependent DNA damage among various groups. In comparison with control group, extensive histological degenerative alterations in the liver, testes, and kidneys were observed in birds of group D and E, where mild to severe alteration like congestion, vacuolation, necrosis, apoptosis, karyopyknosis, extensive degeneration, and alteration in many cellular structures were noticeable.

A new, morphologically distinctive lacertid lizard of the genus Eremias (Rhabderemias) is described from the arid mountains of northwestern Balochistan Province in Pakistan. Eremias kakari sp. nov. has an isolated distribution and can be easily distinguished from all other species of mainly desert subgenus Rhabderemias (E. andersoni, E. cholistanica, E. fasciata, E. lineolata, E. pleskei, E. scripta, E. vermiculata). Apart from other differences, E. kakari sp. nov. can be distinguished from geographically close members of the subgenus Rhabderemias (E. cholistanica, E. fasciata, and E. scripta) by having a single row of subdigital lamellae and a complete row of lateral scales and hence three scales around the penultimate phalanx of 4th toe. The new species is morphologically (dorsal pattern) very similar to E. fasciata but can be distinguished from this species for having 22-26 subdigital lamellae under 4th toe, 48-55 dorsal scales across midbody, ventrals in 11-14 oblique longitudinal series across the belly, 17-21 femoral pores and 17-21 scales in the 9th-10th annulus posterior to the postcloacal granules. The new species is currently known only from the type locality situated in the Toba Kakar Range, near to Tanishpa village. However, we expect that Eremias kakari sp. nov. would have a broader range in northwestern Pakistan and southeastern Afghanistan. An identification key for the Pakistani Eremias, together with other remarks to the new species, is presented.

Members of the dwarf geckos of the genus Microgecko Nikolsky, 1907 are distributed from western Iran to northwestern India, with seven currently recognized species. Three taxa have been reported from Pakistan, M. depressus, M. persicus persicus and M. p. euphorbiacola. The former is the only endemic species restricted to Pakistan. Herein, we describe a new species, Microgecko tanishpaensis sp. nov., on the basis of four specimens collected from the remote area of the Toba Kakar Range in northwestern Balochistan. The type locality lies in an isolated valley in mountainous terrain known for the occurrence of other endemic reptile species, including geckos. Microgecko tanishpaensis sp. nov. is differentiated from the morphologically similar species M. depressus by possessing larger size, five scales bordering the nostril, internasals (supranasals) scales in contact with nostril, two large pairs of postmentals, higher numbers of interorbitals (27-30), scales around midbody (76-84), ventral scales from the postmental to vent (144-156) and scales along dorsal midline from axilla to groin (75-86). A morphological comparison of M. tanishpaensis sp. nov. with other species of the genus and an updated identification key for the genus Microgecko are presented.

Health and environmental problems arising from metals present in the aquatic ecosystem are very well known. The present study investigated toxicological effects of LC 15 of metals such as copper, chromium, and lead for 24, 48, 72, and 96 h on hematological indices, RBC nucleus and cell morphology, and gill and muscle tissues of grass carp ( Ctenopharyngodon idella ). Experimental dose concentrations of copper were 1.5, 1.4, 1.2, and 1 mgL -1 . Similarly, dose concentrations of chromium were 25.5, 22.5, 20, and 18 mgL -1 while those of lead were 250, 235, 225, and 216 mgL -1 , respectively. Maximum decrease in the concentration of Hb, RBCs, and monocytes was observed against chromium, while maximum increase in the concentration of lymphocytes was reported against lead. Abnormalities such as single and double micronuclei, deformed nucleus, nuclear shift, irregular nucleus, deformed cells, microcyte cells, and vacuolated and swollen cells were observed. Gill tissues absorbed maximum concentration of lead followed by chromium and copper. Muscle tissues also absorbed maximum concentration of lead followed by chromium and copper, respectively. Histological alterations such as epithelial lifting, interlamellar spaces, club gill filaments, gill bridging, curling filaments, swelling and fusion of cells, irregular cells, destruction of epithelial cells, cellular necrosis, and inflammatory cells were observed in gill tissues while inflammation and necrosis of muscle fibers, degeneration of muscle fibers, edema of muscle bundles, zig-zag of muscle fibers, and lesions were observed in muscle tissues of fish exposed with different doses of these heavy metals, indicating the toxicity of metals to aquatic fauna as well as to human being via food chain., Competing Interests: The authors declare that they have no conflicts of interest with respect to the work presented here., (Copyright © 2020 Nazish Shah et al.)

Contamination of fresh water bodies like riverine system is utmost concerned environmental issue. This study was aimed to assess the concentration of heavy metals in River Kabul and their bioaccumulation by freshwater mussel. Comet assay was used to evaluate the genotoxic effect of heavy metals on hemocytes of freshwater mussel. The concentration of heavy metals in water was in the order of Zn > Pb > Ni > Cu > Mn > Fe > Cr > Cd, in sediments were Fe > Zn > Cr > Ni > Mn > Pb > Cu > Cd and in the soft tissues of freshwater mussel were in order of Fe > Zn > Mn > Pb > Cu > Cr > Ni > Cd. The hemocytes of mussels from polluted sites showed significantly higher (p < 0.05) DNA damage as compared to reference site. The study showed that pollutants from industries, municipal, domestic and agricultural sources cause heavy metals contamination in River Kabul.

Background: Safety protocols are usually neglected in most of the matchstick industries rendering the laborer prone to various occupational hazards., Objective: The present study highlights DNA damage among matchstick factory workers (n = 92) against a control group (n = 48) of healthy individuals., Methods: Genotoxicity was measured in peripheral blood lymphocytes of the test subjects using a Single Cell Gel Electrophoresis assay (SCGE/comet assay)., Results: Our results substantiate a high Total Comet Score (TCS) for factory workers (74.5 ± 47.0) when compared to the control group (53.0 ± 25.0) (P ≤ 0.001). Age and duration of occupational exposure had no significant effect (P > 0.05) on TCS value. As for job function, the TCS value was greatest in sweepers (91.0 ± 56.1) and lowest in box-making operators (26.0 ± 25.0) indicating that waste disposal poses the higher risk of DNA damage., Conclusions: Our study corroborates that matchstick chemicals can potentially damage the DNA of exposed subjects.

This study was conducted to assess the amount of DNA damage caused by Oxcarbazepine (OXC) through single cell gel electrophoresis (SCGE) technique/comet assay. OXC derived from dibenzazepine series is an effective second generation antiepileptic drug (AED) for both children and adults. Side effects like genotoxic effects of AEDs are of prime importance resulting from toxic metabolites, free radicals and reactive oxygen species (ROS). Forty Eight adult male Bagg's albino mice (BALB/c) were randomly classified into eight groups, each comprising of six animals. Two of these groups were control and six were tested groups. Control groups were injected with 1% tween 80 while tested groups were injected with 10, 20, and 40 mg/kg-day OXC for seven days (acute therapy) and 28 days (subchronic therapy) in peritoneal cavity. Blood samples were collected by cardiac puncture and subjected to comet assay for the analysis of DNA damage. Per sample 100 cells were scored and classified according to comet tail length. The results showed that OXC in acute and long term therapies had significantly higher (p < 0.05) genotoxicity in treated groups as compared to control groups. Our study suggests that OXC may cause significant DNA damage in both acute as well as in subchronic therapies.

Contemporary studies demonstrate that rodent bites do not occur frequently. However, a huge number of cases were reported from Peshawar vale, Pakistan during 2016. Two species, the local black rat Rattus rattus (Linnaeus, 1758) and the invasive brown rat Rattus norvegicus (Berkenhout, 1769) might be the suspected cause. Several studies indicated the invasion of brown rats into Pakistan presumably via port city of Karachi. In this study, we modeled geospatial distribution of rodent bites for risk assessment in the region. Bite cases reported to tertiary care lady reading hospital were monitored from January 1 to August 31, 2016. Among 1747 cases, statistically informative data (n = 1295) was used for analyses. MaxEnt algorithm was employed for geospatial modeling, taking into account various environmental variables (temperature, precipitation, humidity, and elevation) and anthropogenic factors (human population density, distance from roads, distance from water channels, and land use/land cover). MaxEnt results revealed that urban slums (84.5%) are at highest risk followed by croplands (10.9%) and shrublands (2.7%). Anthropogenic factors affecting incidence of rodent bites included host density (contribution: 34.7), distance from water channels (3.2), land use/land cover (2.8), and distance from roads (2). Most of the cases occurred within a radius of 0.3 km from roads and 5 km from water channels. Rodent bite incidence is currently at its peak in Peshawar vale. Factors significantly affecting rodents' bite activity and their distribution and dispersal include urbanization, distance from roads, and water channels. Further studies are needed to determine the impact of invasion by brown rat on bite incidence.

The present investigation aimed to assess the concentrations of selected heavy metals in water and sediments and their bioaccumulation in tissues of freshwater mussels and their histopathological effects on the digestive gland, gills, and gonads of Anodonta cygnea . Water, sediments, and freshwater mussel samples were collected at four sites, that is, reference and polluted sites, along the Kabul River, Khyber Pakhtunkhwa. The polluted sites were receiving effluents from the industrial, agricultural, municipal, and domestic sources. The order of metals in the water was Zn > Pb > Ni > Cu > Mn > Fe > Cr > Cd, in sediments the order was Fe > Zn > Cr > Ni > Mn > Pb > Cu > Cd, and in the soft tissues the order was Fe > Zn > Mn > Pb > Cu > Cr > Ni > Cd. Histopathological alterations observed in polluted sites of Kabul River were inflammation, hydropic vacuolation, and lipofuscin pigments (in digestive gland), gill lamellar fusion, dilated hemolymphatic sinus, clumping, and generation of cilia and hemocytic infiltration (in gills), and atresia, necrosis, granulocytoma, hemocytic infiltration, and lipofuscin pigments (in gonads). The histopathological alterations in the organs of Anodonta cygnea can be considered as reliable biomarkers in biomonitoring of heavy metal pollution in aquatic ecosystems.

The study explored (I) the concentration of heavy metals in water samples (II) their bioaccumulation in common carp Cyprinus carpio (III) and the subsequent genotoxicity in the selected organs of carp; from river Kabul, Khyber Pakhtunkhwa Pakistan. Except for Mercury (Hg) the water samples had all the heavy metals within permissible limits of recommended dietary allowance (RDA). Nonetheless a number of heavy metals (Zn, Ni, Cr, Cd, Pb and Hg) showed bioaccumulation at levels higher than permissible. Zinc (Zn) was the most while Cadmium (Cd) was the least accumulated metal in all tissue samples analyzed. The metal burden in different organs of C. carpio was in sequence of intestine > skin > liver > gills > muscle. The Comet assay established DNA damage in selected organs to be in accordance with metal burden; the most to least damage being in sequence of blood > intestine > skin > liver > gills > muscle. In conclusion assessment of DNA damage in the organs of C. carpio appears to be a useful bio-marker to evaluate genotoxic effects of heavy metal pollution.

Objective: To provide baseline information about suspected vectors and the incidence, distribution and an active zone of transmission for cutaneous leishmaniasis (CL) in Chitral, Pakistan, using GIS tools; and to investigate the role of environmental factors in the disease dynamics., Method: Two surveys in 2014 and 2016 as a basis for choropleth and environmental risk mapping., Results: A total of 769 captured specimens yielded 14 Phlebotomus and six Sergentomyia species including two potential vectors of CL, i.e. Phlebotomus papatasi and Phlebotomus sergenti. P. papatasi (71%) was dominant, followed by P. sergenti (18%). A choropleth map generated in Arcmap 10.1 based on 1560 CL case reports displayed maximum prevalence (0.92-2.5%) in Ayun, Broz, Charun, Chitral 1 and 2 and Darosh 1 and 2 union councils. An environmental risk map constructed by MaxEnt 3.3.3 defined an active zone of transmission based on leishmaniasis occurrence records (n = 315). The analysis of variable contribution in MaxEnt indicates significance of elevation (54.4%), population density (23.3%) and land use/land cover (6.6%) in CL disease dynamics., Conclusion: The probability of CL increases (0.6-1 on logistic scale) in severely deforested areas, in lowland valleys and in regions with high-population density., (© 2017 John Wiley & Sons Ltd.)

North West Pakistan (NWP) is characterized by four eco-zones: Northern Montane Region, North Western Hills, Submontane Region and Indus Plains. Present study identified 1037 cases of traumatic myiasis in the region during 2012-2015. Screw worm larvae were classified as 12 species: Chrysomya bezziana (Villeneuve), Chryomya megacephala (Fabricius), Chrysomya rufifacies (Macquart), Lucilia cuprina (Wiedemann), Lucilia sericata (Meigen), Lucilia illustris (Meigen), Lucilia porphyrina (Walker), Hemipyrellia ligguriens (Wiedemann), Calliphora vicina (Robineau-Desvoidy), Wohlfahrtia magnifica (Schiner), Sarcophaga crassipalpalis (Macquart), Sarchophaga species. Among these C. bezziana, L. cuprina and W. magnifica with approximately 882 case reports were the principal agents of traumatic myiasis. The species W. magnifica is a first report from Pakistan. In order to investigate spatial distribution of these dominant species we used MaxEnt niche model. Our results revealed a well-established occurrence of C. bezziana and L. cuprina in the four eco-regions while W. magnifica is currently contained in the Submontane Region. Several hot spot areas of infestation were detected all characterized by high human population density showing synanthropic nature of these species. Wohlfahrtia magnifica was excluded from Northern Montane Region with severe winters and Southern Indus Plains with harsh summers revealing that invasive species are initially sensitive to extreme of temperatures. Presence of L. cuprina in the wet areas of North Humid Belt (Maximum annual precipitation: 1641mm) depicted a moisture preference of the species. In perspective of changing climate and future predictions of severe events such as droughts and flooding in NWP, W. magnifica can potentially alter the species composition. Considering these findings in an eco-geographically dynamic region of Pakistan we predict that two factors (1) Growing human population (2) Climatic conditions, equally contribute to range shift of synanthropic species., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The aim of the study was to quantify the accumulation of heavy metals like Zn, Ni, Cr, Cu, Cd, Mn, Fe, and Hg in various organs of Wallago attu. Samples were collected from polluted parts of River Kabul and compared with control fish netted from Warsak Dam. The data indicated that the sequence of metal bioaccumulation in the skin, gills, and muscle was Zn > Cr > Pb > Cu > Ni > Fe > Mn > Hg > Cd, in the intestine was Zn > Pb > Cr > Cu > Ni > Fe > Mn > Hg > Cd, and in the liver was Zn > Pb > Cr > Cu > Ni > Fe > Mn > Hg > Cd. The overall metal burden in different tissues of W. attu was in the sequence of skin > gills > intestine > muscle > liver. The skin being the prime target organ showed that the route of metal uptake was the direct result of fish to metal exposure. The liver accumulated the least level of metals than other organs of the same fish.

Herein is reported the first case of urogenital myiasis from Peshawar, Pakistan. Third instar blow fly larvae were recovered from the urinogenital tract of a 5-year old female child. The larvae were identified as Chrysomya bezziana(Villeneuve), using Light and Scanning Electron microscopic techniques. The study brings into focus the subject of human myiasis, about which little is known in Pakistan.

Background: Fuel filling stations workers and automobile workshops mechanics are consistently exposed to gasoline hydrocarbons during their occupation, this may cause DNA damage. Objective of this study was to evaluate the level of DNA damage in subjects occupationally exposed to these hydrocarbons., Methods: Comet assay was performed on blood lymphocytes of exposed subjects to assess the probable DNA damage. 100 cells per individual were scored and graded by comet tail length. Exposed group consisted of 98 subjects (age 25.4 ± 7.2 years), of which 68 were CNG/Petrol filling men and 30 were automobile workshop workers, selected randomly from different service stations and automobile workshops of populated and adjacent cities of Peshawar, Mardan and Nowshera of Khyber Pakhtunkhwa province, while control group included 92 subjects (age 26.7 ± 11.8 years) were also from the same areas., Results: Significantly high level of DNA damage was found in the subjects exposed to gasoline hydrocarbons as compared to control subjects (173.2 ± 50.1 and 61.0 ± 25.0, P = 0.001, respectively). Period of exposure and use of tobacco also showed considerable effects (P < 0.05) on DNA damage, while effect of age and daily working hours on total comet score (TCS) were non-significant (P > 0.05)., Conclusions: The results of our study concluded that petroleum hydrocarbons have the potential to cause DNA damage in the exposed subjects. The study also suggested that protective strategies should be implemented by the concerned authorities to minimize exposure to fuel hydrocarbons.

Objectives: The importance of X-rays as a diagnostic medical tool cannot be denied. However, continuous exposure to X-rays can cause DNA damage. This study aimed to use the comet assay technique to investigate the level of DNA damage in lymphocytes due to X-rays in occupationally exposed personnel., Methods: Blood samples were collected from 74 exposed and 70 control subjects for analysis. A total of 100 randomly captured cells from each slide were examined using an epifluorescent microscope. The comets were analyzed by a visual scoring method according to comet tail length., Results: The results indicated a significant increase (p<0.05) in DNA damage in X-rays technicians (129.8 ± 17.2) as compared with the control group (53.0 ± 25.0). A significant increase (p<0.02) in DNA damage was also observed with an increase in exposure duration of technicians because of their service length., Conclusions: The present study suggests that the exposed radiology personnel should carefully comply with radiation protection procedures such as wearing of lead apron during diagnostic procedures and minimize radiation exposure where possible to avoid potential genotoxic effects due to X-rays.

Associations of GSTT1, GSTM1 and CYP1A1 gene variants with risk of developing oral cancer were evaluated in this study. A case-control study was conducted in Pashtun population of Khyber Pakhtunkhwa province of Pakistan in which 200 hospital based oral cancer cases and 151 population based healthy controls exposed to similar environmental conditions were included. Sociodemographic data were obtained and blood samples were collected with informed consent for analysis. GSTM1 and GSTT1 were analysed through conventional PCR method while specific RT-PCR method was used to detect CYP1A1 polymorphisms. Results were analyzed for conditional logistic regression model by SPSS version 20. The study shows that patients with either GSTM1 or GSTT1 null genotypes have significantly higher risk of oral cancer (adjusted odds (OR): (3.019 (1.861-4.898) and 3.011(1.865-4.862), respectively), which further increased when either one or both null genes were present in combination (adjusted odds (OR): (3.627 (1.981-6.642 and 9.261 (4.495-19.079), respectively). CYP1A1 rs4646903 gene variants individually showed weak association OR: 1.121 (0.717-1.752); however, in the presence of GSTM1 and/or GSTT1 null genotypes further increasing the association (adjusted odds (ORs): 4.576 (2.038-10.273), 5.593 (2.530-12.362) and 16.10 (3.854-67.260 for GSTM/GSTT null and CYP1A1 wild type, GSTM/GSTT either null and CYP1A1 variant alleles, and all 3 gene polymorphisms combinations, respectively). Our findings suggest that presence of GSTM1 and/or GSTT1 null genotypes along with variant alleles of CYP1A1 may be the risk alleles for oral cancer susceptibility in Pashtun population.

The purpose of this study was to evaluate associations of the CYP1A1 gene variant rs4646903 polymorphism with the risk of developing esophageal cancer (EC). A case-control study was carried out in Pashtun population of Khyber Pakhtunkhwa province of Pakistan in which 140 hospital based EC cases and 196 population based healthy controls exposed to similar environmental conditions were included. A specific method based on the real time polymerase chain reaction (RT-PCR) was used to detect genotypes in case and control groups and results were then analyzed with SPSS version 20. In our population, individuals with CC and TC genotypes of the CYP1A1 rs4646903 polymorphism had significantly higher risk of EC (adjusted odds (OR): 15.709, 95%CI: 6.065-40.686, OR: 3.256 95%CI: 1.902-5.574 respectively). The 'C' allele was strongly associated with the disease (p< 0.0001). Adjusted OR was higher (1.5 times in C/C) in case of variant alleles that show the contribution of environmental and nutritional factors towards the development of EC. Our findings suggest that presence of the 'C' allele of rs4646903 (T>C) may be one of the risk alleles for EC susceptibility in Pashtun population.