Exome sequencing identifies homozygous variants in MBOAT7 associated with neurodevelopmental disorder.

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_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_778del;p.Glu253_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.

DNA damage analysis concerning GSTM1 and GSTT1 gene polymorphism in gold jewellery workers from Peshawar Pakistan.

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.

Polymorphism in GSTM1 and GSTT1 genes influence DNA damage in personnel occupationally exposed to volatile anaesthetics (VA), from Peshawar, Pakistan.

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.

Association of Non-Hodgkin lymphoma risk with CYP1A1, GSTM1 and GSTT1 gene variants, in tobacco addicted patients of Pashtun ethnicity of Khyber Pakhtunkhwa, Pakistan.

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.

Bioaccumulation of Heavy Metals and their Genotoxic Effect on Freshwater Mussel.

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.

The genotoxic effect of oxcarbazepine on mice blood lymphocytes.

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.

Rat-bites of an epidemic proportion in Peshawar vale; a GIS based approach in risk assessment.

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.

Bio-monitoring of Tissue Accumulation and Genotoxic Effect of Heavy Metals in Cyprinus carpio from River Kabul Khyber Pakhtunkhwa Pakistan.

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.

Environmental risk modelling and potential sand fly vectors of cutaneous leishmaniasis in Chitral district: a leishmanial focal point of mount Tirich Mir, Pakistan.

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.

Comet Assay and Micronucleus Test in Circulating Erythrocytes of Ctenopharyngodon idella Exposed to Nickel Oxide Nanoparticles.

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.

Unveiling genetics of non-syndromic albinism using whole exome sequencing: A comprehensive study of TYR, TYRP1, OCA2 and MC1R genes in 17 families.

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_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_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.

Genotoxicity of Copper, Silver and Green Synthetic Gold Nanoparticles in Fish (Ctenopharyngodon idella).

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.

Evidence that MEK1 positively promotes interhomologue double-strand break repair.

During meiosis there is an imperative to create sufficient crossovers for homologue segregation. This can be achieved during repair of programmed DNA double-strand breaks (DSBs), which are biased towards using a homologue rather than sister chromatid as a repair template. Various proteins contribute to this bias, one of which is a meiosis specific kinase Mek1. It has been proposed that Mek1 establishes the bias by creating a barrier to sister chromatid repair, as distinct from enforcing strand invasion with the homologue. We looked for evidence that Mek1 positively stimulates strand invasion of the homologue. This was done by analysing repair of DSBs induced by the VMA1-derived endonuclease (VDE) and flanked by directly repeated sequences that can be used for intrachromatid single-strand annealing (SSA). SSA competes with interhomologue strand invasion significantly more successfully when Mek1 function is lost. We suggest the increase in intrachromosomal SSA reflects an opportunistic default repair pathway due to loss of a MEK1 stimulated bias for strand invasion of the homologous chromosome. Making use of an inhibitor sensitive mek1-as1 allele, we found that Mek1 function influences the repair pathway throughout the first4-5 h of meiosis. Perhaps reflecting a particular need to create bias for successful interhomologue events before chromosome pairing is complete.

Appearances often deceive in racerunners: integrative approach reveals two new species of Eremias (Squamata: Lacertidae) from Pakistan.

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.

Assessment of DNA damage induced by endosulfan in grass carp (Ctenopharyngodon idella Valenciennes, 1844).

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.

Metals Distribution, Histopathological Alterations, and Health Risk Assessment in Different Tissues of Fish (Ctenopharyngodon idella).

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.

Out of the blue: The first record of the genus Heremites Gray, 1845 (Squamata, Scincidae) from Pakistan.

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.

Genotoxic Consequences in Common Grass Carp (Ctenopharyngodon idella Valenciennes, 1844) Exposed to Selected Toxic Metals.

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.

Scalp Hair Metal Analysis Concerning DNA Damage in Welders of Peshawar Khyber Pakhtunkhwa Pakistan.

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.

Assessment of copper, chromium, and lead toxicity in fish (Ctenopharyngodon idella Valenciennes, 1844) through hematological biomarkers.

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.