Modelling the occurrence and spatial distribution of screwworm species in Northern Pakistan.

We use binomial kriging to model the spatial distribution of myiasis by three species namely Chrysomya bezziana, Wohlfahrtia magnifica and Lucilia cuprina in the livestock of Khyber Pakhtunkhwa, Pakistan. Traditional species distribution models are usually based on assumption of independence of observations. Species data often come in presence-only form for which background points are generated based on some covariates using statistical and machine learning techniques such as MaxEnt. We assume a symmetric binomial distribution based on the principle of maximum entropy in order to decide the number of pseudo-absences. Our results showed that the spatial models fitted very well and prediction distributions were estimated with excellent accuracy. Moreover kriging maps were more accurate as most of the non-spatial variation has been picked up by external drift with higher values of the sensitivity focusing partial AUC for all the three species. Land-use-land-cover was a common factor significantly affecting spatial distribution of all the three species suggesting that for established species anthropogenic factors such as land use become a strong determinant of their spatial distribution. Our results also revealed that for invading species like W. magnifica elevation acts as a barrier to species dispersal and therefore is more limiting to distribution. Furthermore the higher overall prediction accuracy demonstrated that our models performed well in predicting the distributions of the three species, which would lead to better understanding and management of the larval infestation.

Biomonitoring of DNA Damage in Photocopiers' Workers From Peshawar, Khyber Pakhtunkhwa, Pakistan.

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 ±â€Š60.7) than the controls (56.6 ±â€Š17.2). 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.

Bio-monitoring of DNA damage in matchstick industry workers from Peshawar Khyber Pakhtunkhwa, Pakistan.

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.

Bioaccumulation of Heavy Metals in Water, Sediments, and Tissues and Their Histopathological Effects on Anodonta cygnea (Linea, 1876) in Kabul River, Khyber Pakhtunkhwa, Pakistan.

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.

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.

Spatial modeling of rat bites and prediction of rat infestation in Peshawar valley using binomial kriging with logistic regression.

In this study, we propose to develop a geostatistical computational framework to model the distribution of rat bite infestation of epidemic proportion in Peshawar valley, Pakistan. Two species Rattus norvegicus and Rattus rattus are suspected to spread the infestation. The framework combines strengths of maximum entropy algorithm and binomial kriging with logistic regression to spatially model the distribution of infestation and to determine the individual role of environmental predictors in modeling the distribution trends. Our results demonstrate the significance of a number of social and environmental factors in rat infestations such as (I) high human population density; (II) greater dispersal ability of rodents due to the availability of better connectivity routes such as roads, and (III) temperature and precipitation influencing rodent fecundity and life cycle.

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.

Dataset of traumatic myiasis observed for three dominant screw worm species in North West Pakistan with first report of Wohlfahrtia magnifica (Schiner).

Regional surveys were carried out in different parts of North West Pakistan among domestic animals (N=57,921) including pets and livestock identifying cases of traumatic myiasis (n=1037). A total of four surveys focused general livestock population during Eid ul Adha (Eid surveys; incidence=1.21%) while another four surveys (Miscellaneous surveys; incidence=7.34%) targeted animal population brought to veterinary hospitals and dispensaries. Timeframe spanned four years from 2012 to 2015. Maggots were sampled and location of the wound was recorded for each host. Taxonomic identification used light and electron microscopic techniques. Our dataset shows three species as principle agents of myiasis (n=882) including Chrysomya bezziana Villeneuve (n=394), Wohlfahrtia magnifica (n=244) and Lucilia cuprina Wiedemann (n=244). Others (n=155) including Chrysomya megacephala (Fabricius), Chrysomya rufifacies (Macquart), Lucilia sericata (Meigen), Lucilia illustris (Meigen), Lucilia porphyrina (Walker), Hemipyrellia ligguriens (Wiedemann), Calliphora vicina (Robineau-Desvoidy), Sarcophaga crassipalpalis (Macquart) and Sarcophaga species were identified as species of minor importance. The obligatory screwworm species W. magnifica is a first report from Pakistan. The results based on this dataset are presented in a recent publication "Distribution Modeling of three screwworm species in the ecologically diverse landscape of North West Pakistan" (Zaidi et al., 2016) [1].

Urinogenital Myiasis From Blow Fly in a Pakistani Child.

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.

Distribution Modeling of three screwworm species in the ecologically diverse landscape of North West Pakistan.

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.

Species Distribution Modelling of Aedes aegypti in two dengue-endemic regions of Pakistan.

OBJECTIVES: Statistical tools are effectively used to determine the distribution of mosquitoes and to make ecological inferences about the vector-borne disease dynamics. In this study, we utilised species distribution models to understand spatial patterns of Aedes aegypti in two dengue-prevalent regions of Pakistan, Lahore and Swat. Species distribution models can potentially indicate the probability of suitability of Ae. aegypti once introduced to new regions like Swat, where invasion of this species is a recent phenomenon. METHODS: The distribution of Ae. aegypti was determined by applying the MaxEnt algorithm on a set of potential environmental factors and species sample records. The ecological dependency of species on each environmental variable was analysed using response curves. We quantified the statistical performance of the models based on accuracy assessment and spatial predictions. RESULTS: Our results suggest that Ae. aegypti is widely distributed in Lahore. Human population density and urban infrastructure are primarily responsible for greater probability of mosquito occurrence in this region. In Swat, Ae. aegypti has clumped distribution, where urban patches provide refuge to the species in an otherwise hostile heterogeneous environment and road networks are assumed to have facilitated in passive-mediated dispersal of species. CONCLUSIONS: In Pakistan, Ae. aegypti is expanding its range northwards; this could be associated with rapid urbanisation, trade and travel. The main implication of this expansion is that more people are at risk of dengue fever in the northern highlands of Pakistan.

Utility of multi-gene loci for forensic species diagnosis of blowflies.

Contemporary studies in forensic entomology exhaustively evaluate gene sequences because these constitute the fastest and most accurate method of species identification. For this purpose single gene segments, cytochrome oxidase subunit I (COI) in particular, are commonly used. However, the limitation of such sequences in identification, especially of closely related species and populations, demand a multi-gene approach. But this raises the question of which group of genes can best fulfill the identification task? In this context the utility of five gene segments was explored among blowfly species from two distinct geographic regions, China and Pakistan. COI, cytochrome b (CYTB), NADH dehydrogenase 5 (ND5), nuclear internal transcribed spacers (ITS1 and ITS2), were sequenced for eight blowfly species including Chrysomya megacephala F. (Diptera: Calliphoidae), Ch. pinguis Walker, Lucilia sericata Meigen L. porphyrina Walker, L. illustris Meigen Hemipyrellia ligurriens Wiedemann, Aldrichina grahami Aldrich, and the housefly, Musca domestica L. (Muscidae), from Hangzhou, China; while COI, CYTB, and ITS2 were sequenced for four species, i.e. Ch. megacephala, Ch. rufifacies, L. cuprina, and the flesh fly, Sarcophaga albiceps Meigen (Sarcophagidae), from Dera Ismail Khan Pakistan. The results demonstrate a universal utility of these gene segments in the molecular identification of flies of forensic importance.

A preliminary survey of carrion breeding insects associated with the Eid ul Azha festival in remote Pakistan.

In Pakistan, the three days of Islamic festival of Eid ul Azha (Eid) are characterized by millions of livestock sacrifices and availability of countless breeding resources for insects of forensic importance. Thus during Eid, December, 2008, in a Pakistani village observing 70 sacrifices, we examined postmortem insect attractions on various sacrificial products (from 23 animals), i.e., blood pools, meat, hides/skins, bones, offal and real time insect succession on goats' offal at five dump sites. Thirty five insect species were collected from 14 indoor and outdoor sites. Offal at the dump sites attracted most insect species both by taxa and in numbers, followed by hides in the court yards, meat (40-70 kg/animal) in kitchens and blood pools in the court yards. Synanthropic calliphorids and muscids were found to be exclusive indoor "first and only arrivals" in comparison to a diverse array of first day insect arrivals at dump sites including calliphorids, sarcophagids, muscids, sphaerocerids, sepsids, empidids, gryllids and formicids. Though burial is usually considered as a barricade between most insect species and carrion yet dump site A with an early burial but also earliest insect colonization showed the fastest rate of decomposition. No vertebrate scavenging at any of the offal dump sites perhaps showed their private feeding inclination, thus implying that in times of mass slaughters most of the dumped winter carrion products in human vicinities are either consumed by insects and/or are buried. Here we also suggest a high rate of fly dispersal during Eid which if true has consequences for forensic investigations in Muslim populace areas especially during the Eid.