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The recent emergence of dengue viruses into new susceptible human populations throughout Asia and the Middle East, driven in part by human travel on both local and global scales, represents a significant global health risk, particularly in areas with changing climatic suitability for the mosquito vector. In Pakistan, dengue has been endemic for decades in the southern port city of Karachi, but large epidemics in the northeast have emerged only since 2011. Pakistan is therefore representative of many countries on the verge of countrywide endemic dengue transmission, where prevention, surveillance, and preparedness are key priorities in previously dengue-free regions. We analyze spatially explicit dengue case data from a large outbreak in Pakistan in 2013 and compare the dynamics of the epidemic to an epidemiological model of dengue virus transmission based on climate and mobility data from ∼40 million mobile phone subscribers. We find that mobile phone-based mobility estimates predict the geographic spread and timing of epidemics in both recently epidemic and emerging locations. We combine transmission suitability maps with estimates of seasonal dengue virus importation to generate fine-scale dynamic risk maps with direct application to dengue containment and epidemic preparedness.

Leishmaniasis, an important neglected vector borne disease, endemic in the northern parts of Pakistan. The disease mainly exist in cutaneous form while sporadic cases of visceral leishmaniasis has also been reported from the affected areas. Present study was designed to know the species composition, relative abundance and seasonal variation of leishmaniasis vector in an emerging endemic focus of Nowshera. Sand flies collected by flit method were identified into 4 species of genus Phlebotomus and 17 species of genus Sergentomyia. P. sergenti, the suspected vector of leishmaniasis was also collected in this survey. Sergentomyia baghdadis and S. babu were the most abundant species and sand fly active season extended from March to November with a unimodal distribution and population reaching its peak during the month of August. A supplementary collection from Charun area of Chitral revealed 12 species of Phlebotomus and four species of Sergentomyia. Among the total 31 species collected from both areas, six species of Phlebotomus (P. angustus, P. comatus, P. kabulensis, P. rupester, P. turanicus and P. newsteadi) collected from Chitral and a single species of Sergentomyia (S. sagodina) collected from Nowshera are new reports to Pakistan. Current study also added subgenus Kasaulius to the Phlebotomus of Pakistan. With the addition of these seven species, the checklist of sand flies of Pakistan has increased to 50 species, checklist provided

ABSTRACT Leishmaniasis is endemic for last more than one decade in the tribal district Khyber located in the north-west of Pakistan near Pak-Afghan border. The disease burden has increased during the time of war against terrorism in the tribal belt of Pakistan bordering Afghanistan. This study was designed to investigate various ecological and entomological aspects of sand flies in war effected district Khyber. Sand flies were collected from seven different localities of the district by flit method, CDC light traps and mouth aspirator. A total of 28 species belonging to two genera i.e., Phlebotomus represented by 12 species and Sergentomyia represented by 16 species were collected during the present study. Phlebotomus mongolensis is reported for the first time from Pakistan. Phlebotomus sergenti (48.77%) was the most abundant species followed by Phlebotomus paptasi (17.89%), Sergentomyia baghdadis (14.53%) and Sergentomyia babu (7.22%). Shannon-Weiner Index was high due to maximum number of specimens belonging to Phlebotomus sergenti. The overall female to male sex ratio was observed to be 1:1.9. Sand fly activity was observed from April to December with population density reaching its peak in the month of August. A positive correlation was observed between sand fly abundance and climatic variables, which was significant with temperature only. The high abundance of Phlebotomus sergenti support the endemicity of anthroponotic cutaneous leishmaniasis in the study area. Presence of species of subgenera Adlerius and Larusius suggest the possibility sporadic visceral leishmaniasis in the region.

ABSTRACT Present study aimed at determining the in vitro life cycle of Leishmania tropica. Temporal variability in the morphology and various morphotypes of the promastigotes were noted. Acquisition and propagation of axenic amastigotes was assessed. During promastigote culture, log, mid-log, and late-log phases were observed respectively on day 4, 5 and 6. The stationary phase was observed on day 7. The day following inoculation, most of the promastigotes were nectomonads having long and slender bodies with roughly uniform width from anterior to near posterior end, having as long flagella as one and a half to twice the body. The nectomonads changed to leptomonads with a ratio of 5% and 44% on day 2 and 3 respectively. The body to flagellum ratio decreased in leptomonads giving a wider and shorter appearance than the nectomonads. On day 4, the log phase, the metacyclic promastigotes appeared in the culture. The metacyclic to leptomonad to nectomonad ratio was 27%, 43% and 29% respectively. The ratio of metacyclics steadily increased towards the mid-log (53%), late-log (67%), and stationary phase (83%). Metacyclics had long flagella and short stumpy cell bodies with anterior end much broader than the posterior as compared to that in leptomonads and nectomonads. The posterior end of the body was bearing a slender tail like extension. During logarithmic growth, 17% of promastigotes were found dividing. During a 10 days transformation of axenic amastigotes, the day following inoculation to the day 3 different morphological forms were observed. The day 7 onward only rounded no flagellum (RNF=41-47%) and oval no flagellum (ONF= 53-59%) forms were present. Viability remained 96-98% during transformation. Bulk growth of the promastigotes for prolong (over a week) duration resulted in the acidification of medium and change to viable amastigotes that could be successfully retransformed to promastigotes. Use of orthophosphoric acid for acquiring acidic medium to transform promastigotes to amastigotes p roved effective.

is a significant malaria vector in Pakistan, and understanding its feeding behavior is necessary to control the spread of malaria. However, limited information is available on the host preferences of in Pakistan. Therefore, we aimed to explore the feeding behavior of , a malaria vector, in the District Khyber, Khyber Pakhtunkhwa, Pakistan. A total of 7462 mosquitoes were collected between March and September 2021, with 1674 (22.4%) identified as (952 female and 722 male). Among the female , 495 (52%) were blood-fed. DNA was extracted from the blood-fed female mosquitoes using the Ammonium Acetate Precipitation Method followed by PCR analysis, blood meal sources were identified. Nested PCR on 191 pooled samples was used to detect and . Cattle blood meals were predominant (73%), followed by human (20%) and chicken (7%), with no dog blood meals detected. All individual mosquito samples were negative for , while two pooled samples (out of 191) tested positive for . in Khyber District primarily displayed anthropophagic feeding behavior, with a small portion of the population infected with . The results underscore the importance of targeted vector control strategies, environmental management, community engagement and continuous monitoring to suppress malaria transmission.

ABSTRACT Taxonomically ants are the most neglected group of insects to be studied in Pakistan. The current study was conducted to collect ants from all three tehsils: Charsadda, Tangi and Shabqadar of district Charsadda. Ants were collected randomly from indoor(rooms, kitchen and washrooms) and outdoor(streets, crop fields, barren ground and grave yards) sites by using buccal aspirator, pitfall traps and hand pick method. A total of 19 species representing 11 genera and three subfamilies namely Formicinae, Myrmicinae and Dolichoderinae were identified during the present study. Myrmicinae was the most diverse subfamily with five genera(Crematogaster, Meranoplus, Messor, Monomorium and Pheidole) and 11 species(Crematogaster subnuda, Meranoplus bicolor, Messor instabilis, Monomorium aberrans, Mo. dichroum, Mo. indicum, Pheidole binghami, Ph. fergusoni, Ph. indica, Ph. latinoda, Ph. sulcaticeps). Fromicinae was presented by seven species placed in five genera: Camponotus having three species(C. compressus, C. oblongus and C. sericeus) and one species each of Cataglyphis(Ca. setipes), Lepisiota(L. frauenfeldi), Paratrechina(P. longicornis) and Polyrachis(Po. hauxwelli) while subfamily Dolichoderinae was represented by a single species i.e. Tapinoma melanocephalum. The current study reports one subfamily(Dolichoderinae) seven genera(Cataglyphis, Cremotogaster, Monomorium, Messor, Pheidole, Polyrachis and Tapinoma) with 13 species representing new country records for Pakistan.

Cutaneous Leishmaniasis is endemic in the tribal district of Khyber near the Pak-Afghan border and is caused by Leishmania tropica . In Pakistan, cutaneous leishmaniasis caused by L. tropica is considered anthroponotic and is thought to be maintained by a human-sand fly-human transmission cycle. Along with humans, other mammals may also be acting as reservoir hosts of leishmaniasis in the study area. To investigate the role of non-human mammals in the transmission of leishmaniasis, blood samples were collected from 245 animals from the CL endemic district of Khyber, Pakistan. Leishmania parasite in these samples was detected by amplifying the species-specific sequences in minicircle kinetoplast DNA, using PCR. L. tropica DNA was detected in 18 (7.35%) samples, comprising 11 cows ( Bos taurus ), 6 goats ( Capra hircus ), and 1 dog ( Canus lupus familiaris ). Only a single cow and dog had a leishmaniasis-like lesion, and the remaining positive samples were asymptomatic. None of the tested sheep ( Ovis aries ) and rat ( Rattus rattus , Rattus norvegicus) was positive. The present study reports the first instance of molecular detection of L. tropica in domestic animals. Our study indicates that along with humans’ cows, goats and dogs may also be playing an important role in the transmission of anthroponotic cutaneous leishmaniasis in district Khyber in particular and Pakistan in general.

Background: Anopheles stephensi is a significant malaria vector in Pakistan, and understanding its feeding behavior is necessary to control the spread of malaria. However, limited information is available on the host preferences of A. stephensi in Pakistan. Therefore, we aimed to explore the feeding behavior of A. stephensi, a malaria vector, in the District Khyber, Khyber Pakhtunkhwa, Pakistan. Methods: A total of 7462 mosquitoes were collected between March and September 2021, with 1674 (22.4%) identified as A. stephensi (952 female and 722 male). Among the female A. stephensi, 495 (52%) were blood-fed. DNA was extracted from the blood-fed female A. stephensi mosquitoes using the Ammonium Acetate Precipitation Method followed by PCR analysis, blood meal sources were identified. Nested PCR on 191 pooled samples was used to detect Plasmodium falciparum and Plasmodium vivax. Results: Cattle blood meals were predominant (73%), followed by human (20%) and chicken (7%), with no dog blood meals detected. All individual mosquito samples were negative for Plasmodium falciparum, while two pooled samples (out of 191) tested positive for P. vivax. Conclusion: A. stephensi in Khyber District primarily displayed anthropophagic feeding behavior, with a small portion of the population infected with P. vivax. The results underscore the importance of targeted vector control strategies, environmental management, community engagement and continuous monitoring to suppress malaria transmission.

During the span of August–October, 2017 a major outbreak of Dengue fever happened in Khyber Pakhtunkhwa province of Pakistan. Cases were reported from all the major cities and rural areas, but Peshawar was more severely hit with more than half of the total cases belonging to central Peshawar city. The epidemic patterns reveal that dengue fever cases were mostly reported for plain areas and also low altitude mountainous regions. We employed the principle of maximum entropy to establish the underlying distribution of dengue presences and background data. A geostatistical analysis was conducted by modelling the spatial structure of the dengue fever risk and estimating the prediction maps with corresponding uncertainty taking into account some of the most significant covariates. The prediction maps were created using binomial kriging with a binary logistic drift. The analysis was carried out for the whole province as well as subregions to have a closer look of the spatial distribution at local level. Our results show that our methodology performed well. Vector distribution, population density, and distance to roads were found to significantly affecting the spatial distribution of risk and gives very informative pattern.