Identification of Lernaea Species in Icthyofauna in District Mardan KPK, Pakistan

Identification of Lernaea Species in Icthyofauna in District Mardan KPK, Pakistan | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Identification of Lernaea Species in Icthyofauna in District Mardan KPK, Pakistan Ashfaq Ahmad This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4127646/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Lernaea cyprinacea, a parasitic cyclopoid copepod, is found to parasitize in the brood stock, nursery and culture system in the freshwater hatchery of Mardan regions. The fish were severely infected on their body surface showed dull and sluggish movements. This research, carried out in the district Mardan Khyber-Pakhtunkhwa, Pakistan, spanning from February to August 2023, delves into the prevalence and infestation dynamics of Lernaea parasites in freshwater fish in river Kalpani. Through a comprehensive analysis of 159 specimens collected from various aquatic sources, the study identifies the presence of five parasite species, notably with Saprolegnia as the predominant one. Utilizing a categorization method, the investigation discerns varying degrees of infestation, underscoring the susceptibility of local fish populations to Lernaea. These findings corroborate with earlier research, accentuating the pressing need for targeted mitigation strategies and robust conservation initiatives. Additionally, the study unravels the profound impact of Lernaea cyprinacea on 103 fish, emphasizing its pivotal role as a primary cause of fish mortality. This research not only contributes crucial insights for shaping effective conservation measures but also proves imperative for sustaining the overall health and biodiversity of freshwater ecosystems in the region. Lernaea hatchery Mardan Pakistan Prevalence Kalpani Parasite and Fish Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 BACKGROUND A variety of problems is caused by parasites in larger organisms which may lie inside or outside of the body during its life cycle (Llewellyn et al., 2014 ). The major groups of ectoparasites are Arthropods (crustaceans), Platyhelminthes (monogenesis), and Protozoans. The protozoans are mostly commensal on fish bodies and free living. Most of it causes harmful diseases and is lethal for the host. Ciliates are one of the most common ectoparasites (Buchmann et al., 2001 ). Lernaea are the copepod parasite which is known as an Anchor worm which may cause infestation and death in a variety of freshwater fish fauna. It may infect the wild species as well as the pond fish. The favorable conditions for the infestation of Lernaea species are the summer season and slow-moving water (stagnant water body). About 110 (hundred and ten) species of the parasite (Lernaea or similar to Lernaea) have been explored up to now. The Lernaea cyprinacea are members of Lernaeid which are found throughout the world. The parasite is common in Cyprinids goldfish, grass carp, and common carp. It can infect the catfish which are mostly stocked in ponds with bighead carp. The Lernaea is the crustacean and copepod that also infects shrimp, crabs, and lobsters in various stages of its life cycles, unlike other free-living copepod species which may not cause disease in the host. The parasites of this group have a complex life cycle, the individual of the group is not passed by the help of an intermediate host organism; whereas it can spread directly from one fish body to the other. The organism needs only a single fish (a member of the class amphibian) to develop from an egg to an adult. The total life cycle of the parasite is completed in 18 to 25 days (Robert et al ., 2001). The host is the fish which are infected by the parasites. Some parasites have suckers for attachment to the host body while the rest of them lay eggs. It can modify its immunity and biochemistry according to the situation, and cannot be killed or digested. The economic losses occur in the Fisheries sector of Pakistan due to infection of Lernaea species, monogenic species, Argulus species, and Dactylogyrus species (Daulat et al ., 1976). Productivity and fish health are affected by parasites among the ectoparasites. The fish migration and behavior patterns are affected by parasitism. The community structure, survival value, and reproduction of the host are negatively affected by the parasites (Luque and Poulin 2008 ). Some of the parasites become pathogenic and lethal for fingerlings and fry while most of them show commensalism (Demopoulos et al ., 2015). Lernea Cyperaceae Linnaeus is an introduced fish parasite that can cause mortality in various habitats including fish hatcheries and ponds, the species can mostly affect high-density areas with higher temperatures (zman and Rayner 1958). The parasite Laernaea cyprineace is most abundant and harmful among the members of the family Larnaea. (Bilal et al., 2021 ). The Lernaeid are the fish parasites that belong to the main group of Crustacea (Copepoda). An extensive morphological fitness (adaptation) is shown by the Lernnaeidae family of cyclopoids which keeps them a little bit distinct from the members of the genus Cyclops and others (Ozel et al .,2014). The parasite was first found in Asia later it may be shifted to another continent of the world with farm fish (Innal and Avenant, 2012). The parasite is recorded from various regions of the world including Asia, Southern Africa, Australia, Europe, and its adjacent areas (Hoffman, 2019 ). The Lernaea cyprinacea is found in Nurseries, brood stocks water hatcheries, and fish ponds of Jessore regions. The parasite ( Lernaea cyprinacea ) may infect perches, catfish, Minor barbs, and major carps (both Chinese and Indian carps) (Hossain et al., 2018 ). A diverse habitat is occupied by individuals of the family Lernaeidae, parasites are found in marine and freshwater. The family is represented by fourteen (14) genera with one hundred and ten (110) species (Ho, 1998 ). The parasites are more in number in the aquatic ecosystem and survive with their host in a dynamic equilibrium (Marcogliese, et al ., 2011). Human health is directly or indirectly affected by infectious. The allergic reaction is caused by either pathogen or living parasites present in the tissues of fish bodies (Buchmann et al., 2001 ). The adult member of Lernaea cyprinacea attached to the skins and muscles of the host leaves an opening to enter the secondary microbe and cause death to the infected fish (Van Duijn et al ., 1973). The pathogenicity of lernaeids largely depends on the size of their host and attachment site preferences. Aggregated infections of the mouth and lips occur in Barbus altianalis with Lernaea barinmiana and in cichlids by Lernaea cyprinacea, fin infections caused by Lernaea lophiara in cichlids (Fryer, 1968 ; Paperna, 1996 ). Fish with large numbers of parasites will be thin and their body color becomes darkened, also extensive abrasions appear with loosing and sloughing on its skin. Severe destruction of the gills occurs as well as losses and mortalities up to 50% (Eissa et al ., 2004). The life cycle of the parasite Lernaea has nine stages, out of which three stages are free-living naupliar, five copepodid, and one stage of an adult. During the fourth stage, copulation of both adult males and females occurs at the first host. When the copulation insures the male is dyed whereas the female metamorphoses and enters the anterior tissues of the host body, laying eggs (Nagasawa et al., 2007 ). A tremendous modification in cephalothorax has been shown by the adult female of the genus Lernaea. The body is vermiform and elongated, and the mouth of the parasite female is much reduced. The head is changed into 4 horn shaped compartments which are now cylindrical and long, the anterior appendages are simple while the two posterior ones are bifurcated. The parasites use the horns for attachment to the host body and it’s buried under the epidermis. The abdomen is bent toward the dorsal and short in length. The egg sacs protrude beyond the body while the genital pore is present near the posterior end of the body. The immature forms of both sexes (male and females) and an adult male are free-living; whereas the mature or an adult female of learned have the capability of parasitism. Lernaea cyprinacea attacked massively with the highest mortality and pathogenicity, mostly in the season of summer. The parasite caused ulceration and hemorrhages along with penetration (Carnevia et al ., 2003). The native as well as introduced species of fish fauna in the South American continent are parasitized by 3 species of Lernea, (Boxshall et al., 1997 ). The parasite was recorded in the fish fauna of Argentina, Brazil, Colombia, Chile, and Uruguay (Habit et al ., 2015). The harmful effects of the concerned parasite were observed both in wild and cultured fish populations (Yanong et al., 2002 ). The species causes serious economic loss in various countries of the world (Rogers, 1979 ). Lernaeosis is a major disease that is caused by the parasite in the hosts (Hua et al., 2019 ). The parasites may cause huge destruction and dangerous diseases in fish fauna, may lead to the extinction of species, and can damage the health of humans by consuming them as food (Habib et al., 2019 ). Economic losses have been observed in various formed fishes due to lernaeid ectoparasites globally (Kir et al ., 2007). Lernaeid copepods are considered one of the harmful parasites of freshwater ecosystems; a few individuals can kill young fish (Kabata et al ., 1987). AIMS AND OBJECTIVES To find the prevalence of Lernaea species in the study area. MATERIALS AND METHODS Study Area The study was conducted in District Mardan, river Kalpani, ponds of Tehsil Takht Bhai and Shergarh Khyber Pakhtunkhwa Pakistan. District Mardan lies between 34.1986˚ N, latitude and 72.0404˚ E longitude with a total area of 1632 km 2 . Tehsil Takht Bhai lies between 34.3314˚ N, Latitude, and 71.9046˚ E, longitude with a total area of 353.0 km 2 . Shergarh is a city of Tehsil Takht Bhai which lies between 34.39349˚ N latitude and 71.8996˚ E longitude. Figure 3.1: Study area and data collection center for Mardan District (figures were altered using Google.com) Fish sample collection Between February to August 2023, fish samples were collected from 9 ponds of District Mardan, Tehsil Takht Bhai, Shergarh, and river Kalpani Khyber Pakhtunkhwa Pakistan. Fish samples were tallied to determine whether they were positive or negative for Lernaea infection. The number of Lernaea on each fish was counted in positive samples to evaluate the degree of infestation. Fish sample processing The collected fish samples from the study site were brought immediately in oxygenated bags to the lab, department of Zoology GDC Lundkhwar. Morphometric measurement was done by taking total length [TL] fork length [FL] standard length [SL] anal length [AL] pre orbital length [POL] post orbital length [POL] head length [HL] and body depth [BD]. The fish were weighed by using a digital scale and species identification was done by using the method of Hoffman, 2019 . Preliminary Treatment of Fish The fishes were first given serial numbers, and then the length [in cm] of the fishes was measured from the tip of the snout to the end of the caudal fin using a measuring scale. The weight was taken by using a digital weighing scale. The external body surface and the structures, notably, the skin and fins were thoroughly examined for the presence of lernaea parasites. Skin The scales of the scaly fishes were removed softly with the help of a scalpel. They were cleaned thoroughly with running water and then with distilled water to ascertain the presence of lernaea ectoparasite. Mucus and epithelial tissue are also gently scraped wherever necessary. The materials were then transferred to a small drop of water on a glass slide and covered with a small size cover glass. The materials were then examined under a microscope and recorded. Fins The fins were examined under the microscope after they were cut off and transferred to a Petridis with water. Epithelial scrapings from the fins were made and subjected to microscopic examination. Parasitological Study For parasite collection and identification, the fish samples were deeply examined with naked eyes after that they were examined with a hand lens; naked eye parasites including lernaea species were removed by using forceps and preserved in 70% ethanol. For microscopic ectoparasite, examination was conducted using the method of scraping (Kabata, 1985). For further studies, the isolated parasites were collected, counted, and processed following Gussev (1986). Morphological Identification Lernaea was taken from the fish body using tweezers, rinsed with physiological NaCl, and preserved with lacto phenol. The Lernaea samples used for identification were preserved in absolute ethanol. Lernaea was observed in the holdfast, abdomen, position, and number of legs. The body and anchor length were measured. Lernaea was identified using Kabata’s [15] identification key. RESULTS This research aimed to assess the prevalence, intensity, and abundance of Lernaea species within the aquaculture population in the district of Mardan, Pakistan. Over a period of six months, from February to August 2023, a total of 159 fish samples were collected. These samples were sourced from both the Kalpani River and a hatchery in Shankar Mardan. The examination for lernaeid parasites was conducted in the laboratory of the Department of Zoology at GDC Lundkhwar. The overall prevalence of Lernaea species disease in Dist. Mardan aquaculture was found as 64.77%. The prevalence of Lernaea species disease in where much positive in the selected samples summarized in Table 4.1 . Table 4.1 Shows the total prevalence along with positive and Negative samples Sr. No Length Total samples Positive Negative Positive %age Negative %age Total P-Value 1 10-60cm 114 74 40 64.91 35.08 64.77% 2 81-90cm 45 29 16 64.44 35.55 A total of 114 samples were collected from fish ranging from 10 to 60 cm in length. Among these, 74 samples tested positive, indicating a frequency of 64.91%. Additionally, 45 samples were obtained from fish measuring 81 to 90 cm in length, with 29 of them showing positive results, resulting in a prevalence of 64.44%. The above values are graphically explained in Fig. 4.1 . For fish within the 10–20 cm size range, a maximum of 29 samples were collected, with 15 of them yielding positive results, indicating a positivity rate of 51.72%. In the 21–40 cm size category, 35 samples were obtained, and 28 of them tested positive, resulting in a higher positivity rate of 80%. Among the 41–60 cm size group, 50 samples were collected, with 31 showing positive results, yielding a positivity rate of 62%. Lastly, in the 81–90 cm size range, 45 samples were taken, and 29 of them tested positive, indicating a prevalence value of 64.44%. Table 4.2 Length-wise distribution of samples and their disease prevalence value Sr. No Length Total Samples Positive Negative Prevalence value 1 10–20 cm 29 15 14 51.72 2 21–40 cm 35 28 07 80 3 41–60 cm 50 31 19 62 4 81–90 cm 45 29 16 64.44 The distribution of Lernaea cyprinacea on various parts of Nile Tilapia (Oreochromis niloticus) is illustrated in Fig. 4.4 . The figure reveals that the area around the base of the dorsal fin exhibited the highest infestation by these copepods, with a recorded presence of 78 instances of Lernaea cyprinacea. A collection of 159 specimens of freshwater fish fauna was gathered and identified, encompassing seven distinct species. Within the studied area, a total of five parasite species were observed in association with these fish. The parasite species were Carpora arthritis , Saprolegnia , Spherosporaauretus , Lernaeacyprinacea , and Microsporadia . Table 4.3 Parasite species distribution Name Carpora arthritis Saprolegnia Spherosporaauretus Lernaeacyprinacea Microsporadia No of species 25 70 14 23 27 We utilized a categorization system that classifies infestation levels as light (1–5 Lernaea per fish), moderate (6–10 Lernaea per fish), or heavy (more than 10 Lernaea per fish). Table 4.4 Lernaea per fish degree of infestation. Sr. No Lernaea per fish No of infected sample percentage 1 1–5 32 20.12 2 6–10 52 32.70 3 more than 10 75 47.16 A total of 159 fish samples were gathered from various locations within District Mardan. Specifically, 45 samples were collected in Takht Bhai, 49 samples in Shergarh, and 65 samples were obtained from River Kalpani, as detailed in Table 4.5 . Table 4.5 Area wise lernaea parasite prevalence Sr. no Location Number of Samples Positive Positive Percentage 1 Takhtbhai 45 32 71.11 2 Shergarh 49 17 34.69 3 River Kalpani 65 23 35.38 DISCUSSION Aquaculture stands out as one of the globe's most economically significant sectors, and fish, being a crucial nutritional resource, holds immense value for humanity. The socio-economic development aimed at fostering healthy ecosystems and ensuring the survival of life on Earth necessitates a concerted effort to safeguard water resources and the ecosystems connected to them. The study conducted from February to August 2023 aimed to evaluate the prevalence and infestation levels of Lernaea parasites in fish. Sample collection took place in different ponds near the Takhtbhai archaeological site, spanning District Mardan, Tehsil Takhtbhai, Shergarh, and the Kalpani River in Khyber-Pakhtunkhwa, Pakistan. A total of 159 specimens of freshwater fish fauna were collected and identified, representing seven distinct species. Within the specified area, a total of five parasite species were identified in association with these fish. The parasite species were Carpora arthritis , Saprolegnia , Spherosporaauretus , Lernaeacyprinacea and Microsporadia . The saprolignia was more frequent than the rest of four parasites. The parasites infect the whole body of the host including the gills, head and skin. The all entire system of some hosts was badly infected by the parasites. Khan et al., ( 2023 ) collected Seventy-six (76) specimens of fish fauna were collected from seventeen (17) ponds during the period from September to November 2021. The sampling locations encompassed various districts, including Swabi, Mardan, Charsadda, and Peshawar in the Khyber-Pakhtunkhwa province. The aim of the study was to investigate the ectoparasites associated with the fish fauna in these regions. The outcomes of our study unveil a significant prevalence of Lernaea infestation within the fish populations in our designated study area. Following the categorization method introduced by Kriswijayanti (2019), our assessment differentiated infestations into three levels: light (1–5 Lernaea per fish), moderate (6–10 Lernaea per fish), and heavy (more than 10 Lernaea per fish). The findings indicate varying degrees of susceptibility to Lernaea infestation among the fish in the Takhtbhai region. Our observations align with previous research by Khan et al., ( 2023 ), who reported a similar prevalence of Lernaea in fish populations in neighboring regions. Our study highlights the crucial need to comprehend the presence and impact of Lernaea parasites to facilitate effective conservation endeavors. Notably, it emphasizes the necessity for targeted mitigation strategies, especially for fish populations experiencing heavy infestations. This emphasis on tailored strategies aligns with the conservation recommendations put forth by Ahmed and Rahman (2017) in their research on parasite management within local aquatic ecosystems. About forty three (43) species of fishes and three (species) of amphibians were identified up to species level in the area which were in infected by Lerneacyprinacea . Charciform was more parasitized among the rest of seven (7) orders in the continent. Ahnelt et al., ( 2018 ) reported that a variety of fish fauna is infected by the parasitic copepod Lernaeacyprinacea . Ectoparasite infestation stands out as a primary contributor to mortality among infected fish species. The prevalence of the Lernaea cyprinacea parasite is notably higher in pelagic species compared to smaller benthic fish species. Tayyab (2017) conducted a study on the River Indus, aiming to investigate parasite infestation within the freshwater ecosystem of the specified study area. The prevalence of Lernaea species was found to be significant, accounting for 39.47% of the observed parasites in the fish fauna. This species demonstrated higher lethality compared to other parasite species. Following Lernaea, Gyrodactylus and Dactylogyrus species constituted 19.73%, Icthyopthyrius multifilis species accounted for 17.10%, thelohanellus species represented 11.84%, Loxophyllum species constituted 10.52%, and Piscinoodinium pillulare species made up 7.89% of the observed parasites. All seven species collected during this study were identified for the first time in the study area. According to Innal et al ., (2020), Lernaea cyprinacea , commonly known as anchor worm, is prevalent in a wide range of hosts and belongs to the genus Lernaea. This species has been documented in various studies in the Mediterranean region, specifically in Turkey. The region encompasses a diverse group of cypriniforms, and the parasites were identified in both native and introduced fish populations within the area. FINDINGS This investigation was conducted in Takhtbhai area, Khyber-pakhtunkhwa, Pakistan, from Feb. 2023-July 2023. The purpose was to know the number of Lernaean parasites living and the level of infestation in freshwater fish. One hundred and fifty-nine specimens belonging to twelve members of seven species of fish from different ponds and the Kalpani River were collected. The result of this research, Trematoda found 5 species of which Saprolegnia were the most abundant. Infected juveniles were observed at higher and lower levels, and the study employed a categorization method, where the degree of severity was noted to determine the difference in the susceptibility of fish to Lernaea infestation. The outcome correlate with past findings meaning highlighting how necessary the required counter-attack techniques and the great conservation solutions is, especially in those fish heavily infected by Lernaea. This research has also been valuable because it has demonstrated the richness of fish and amphibian species in the area, the fact that all 43 of the fish species are infected by Lernaea cyprinacea. A study found that Lernaea infestation in addition to other external parasites was the main factor leading to the fish mortality. Lernaea cyprinacea was revealed as the most commonly occurring parasite whereas in comparison the size of the other organisms was smaller. This research highlights how parasitic infestations of freshwater ecosystems may affect the ecologic dynamics, and the parasitic infestations should be considered in management plan in the region to conserve the system effectively CONCLUSION As a conclusion, this research analysis for district Mardan region discourages the intensive infection of the louse fish parasite within the freshwater fishes and underlines that applicable unsustainable measures are key to remedying the infestation varying from low to grave level. The conclusions provide the evidence of high vulnerability of these fish populations to Lernaea infestation and can be compared with previous studies, negative implication can be made that appropriate conservation measures should be adopted. The data collected through this pilot investigation regarding the different types of fish and amphibians that suffer Lernaea cyprinacea infestation are important to the greater ecological picture on how ectoparasites in freshwater can affect the balance. The summarized study facilitates the accumulation of new helpful information with formulation of conservation strategies that promote the wellbeing and sustainability of water bodies in the Khyber-Pakhtunkhwa province, Pakistan. Abbreviations KPK : Khyber Pakhtunkhwa GDC : Govt Degree college TL : Total length FL : Fork length SL : Standard length AL : Anal length POL : Pre orbital length HL : Head length BD : Body depth Declarations the Ethical Review Committee (ERC) of Abdul Wali Khan University Mardan has granted ethical approval for this study. 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Zoological Stud 43(2):193–205 Piazza RS, Martins ML, Guiraldelli L, Yamashita MM (2006) Parasitic diseases of freshwater ornamental fishes commercialized in Florianópolis. Santa Catarina, Brazil Boletim do Instituto de Pesca 32: 51–57 Robert B, Jackson SR (2001) Water in a changing world. Ecol Appl 11(4):1027–1045 Rogers WA (1979) Principal Diseases of Farm-Raised catfish. Helmintic Parasite South Coop Ser 225:40–50 Sayyadzadeh G * 1 ; Esmaeili H.R.1 ; Ghasemian S.1 ; Mirghiyasi S.1 ; Parsi B.1 ; Zamanpoore Akhlaghi M2 (2016) M.3. Co-invasion of anchor worms Lernaea cyprinacea (Copepoda: Lernaeidae) in some freshwater fishes of the Kor River Basin, Southwest of Iran with some remarks on the ecological aspects of lernaeosis in the country. Iranian Journal of Fisheries Sciences 15(1) 369–389 Silva-Souza A, Almeida T, Machado S (2000) Effect of the infestation by Lernaea cyprinacea Linnaeus, 1758 (Copepoda, Lernaeidae) on the leucocites of Schizodon intermedius Garavello and Britski, 1990 (Osteichthyes, Anostomidae). Revista Brasileira de Biología 60(2):217–220 Tayyab M, Gul S, Rehman UH, Rehman UA, Saeed K, Khan S, Naz N, Ahmad A (2017) J Entomol Zool Stud 5(4):422–426 Tekin-Özan, Kir SI, Barlas M (2008) Helminth parasites of common carp. Cyprinus carrpio L., 1758) in Beyșehir lake and population dynamics related to month and host size. Turk Fish J Aquat Sci, 8: 201–205 Waicheim AM, Arbetman M, Rauque C, Viozzi G (2019) The invasive parasitic copepod Lernaea cyprinacea: updated host-list and distribution, molecular identification and infection rates in Patagonia. Aquat Invasions 14(2):350–364 Woo PTK (2002) Fish Diseases and Disorders. CABI International Wallingford Oxfordshire OX10 8DE, UK. 808 pp Woo PTK (1995) Fish diseases and disorders. CAB, Int. Wallingford, Oxon, Uk Van Duijn J (1973) Diseases of fishes Illffe books London Yanong RP, Francis-Floyd R, Curtis E, Klinger RE, Cichra MF, Berzins (2002) I. K. Algal dermatitis in cichlids Journal of the American Veterinary Kriswijayanati BD, Kismiyati K, Kusnoto K (2019) Identifikasi dan derajat infestasi Lernaea pada ikan maskoki (Carassius auratus) di kabupaten Tulungagung, Jawa Timur (Identification and degrees of Lernaea infestation in Goldfish (Carassius auratus) at Tulungagung, East Jawa). J Aquac Fish health, 3(1): 1–7 Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Ahmad","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/ElEQVRIiWNgGAWjYFACHoYPUBb75x8VQIqZuYGQFsYZUBYbM8MZkBZGUrQwtoFoAlr4+88ebPhRZhfNz9777HHhvNpo/naglh8V23BqkbiRl9jYcy45d2bPcXPjmduO5844zNjA2HPmNm5rbvCYP+BtY87dcCONQYJ327HcBqAWoAtxa5E/f8aw8W9bfe7++8+AWuYcy51PSIvBgRzDZt62w7kbJNjYpHkbanI3ENJieAOoReYc0Atn0pgNZxw7kLsRqOUgPr/IgRz2pqw6t7/9GOODDzV1ufPOHz744EcFHu+DARucdRhMHiCgHkVLHWHFo2AUjIJRMOIAANBsYTEbAEANAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0003-2556-9263","institution":"Abdul wali khan University","correspondingAuthor":true,"prefix":"","firstName":"Ashfaq","middleName":"","lastName":"Ahmad","suffix":""}],"badges":[],"createdAt":"2024-03-19 06:06:54","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4127646/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4127646/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":55102683,"identity":"a9dd7c5d-bf7f-4d4b-88a4-02a77e093588","added_by":"auto","created_at":"2024-04-22 15:51:57","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":48388,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 3.2 Study Area\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Picture1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4127646/v1/16c71b4ac01535dafa0afc7c.jpg"},{"id":55101972,"identity":"98b7c8ff-4147-4715-8b30-f1d7de425ca2","added_by":"auto","created_at":"2024-04-22 15:35:57","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":127699,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 3.2 Study Area\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Picture2.png","url":"https://assets-eu.researchsquare.com/files/rs-4127646/v1/9728860d0a980da11f69141f.png"},{"id":55101973,"identity":"537f59d9-a714-4da7-8044-6039daec058d","added_by":"auto","created_at":"2024-04-22 15:35:57","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":154053,"visible":true,"origin":"","legend":"\u003cp\u003eFigure 4.1 Lernaea Positive Fish Samples\u003c/p\u003e","description":"","filename":"Picture4.png","url":"https://assets-eu.researchsquare.com/files/rs-4127646/v1/3d0406ab05da12d80d04377d.png"},{"id":55101974,"identity":"98d570ab-b30a-4de5-b9a1-e07ca3030fee","added_by":"auto","created_at":"2024-04-22 15:35:57","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":209003,"visible":true,"origin":"","legend":"\u003cp\u003eFigure 4.2 Lernaea Negative Fish Samples\u003c/p\u003e","description":"","filename":"Picture5.png","url":"https://assets-eu.researchsquare.com/files/rs-4127646/v1/9a28e1149c93e1c6b181747d.png"},{"id":55101977,"identity":"6e4d41a5-d5a2-404c-8d29-b7f56192b260","added_by":"auto","created_at":"2024-04-22 15:35:57","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":244497,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 4.3 Length-wise distribution.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Picture6.png","url":"https://assets-eu.researchsquare.com/files/rs-4127646/v1/fd3c1a5e4904ecb99f2d9d5b.png"},{"id":55102293,"identity":"b03a6066-5938-46a0-a718-f035b81c9bc3","added_by":"auto","created_at":"2024-04-22 15:43:57","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":132347,"visible":true,"origin":"","legend":"\u003cp\u003eFigure 4.4 Distribution of Lernaea cyprinacea on different parts of the body\u003c/p\u003e","description":"","filename":"Picture7.png","url":"https://assets-eu.researchsquare.com/files/rs-4127646/v1/76c492619442a819cd9bf03b.png"},{"id":55102291,"identity":"598f38e4-ddcb-4348-b52a-ab976a959377","added_by":"auto","created_at":"2024-04-22 15:43:57","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":198847,"visible":true,"origin":"","legend":"\u003cp\u003eFigure 4.5: Parasite species distribution\u003c/p\u003e","description":"","filename":"Picture8.png","url":"https://assets-eu.researchsquare.com/files/rs-4127646/v1/b27c8b6b027b0fa255135180.png"},{"id":55101982,"identity":"b8804391-8a0f-4344-9e30-25ff2adf3417","added_by":"auto","created_at":"2024-04-22 15:35:58","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":288756,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 4.6: Lernaea per fish degree of infestation.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Picture9.png","url":"https://assets-eu.researchsquare.com/files/rs-4127646/v1/858a54bb2e597db1aa0601da.png"},{"id":55101980,"identity":"bf36fb78-923f-44f3-a3bb-0e1b648e4cc3","added_by":"auto","created_at":"2024-04-22 15:35:58","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":294000,"visible":true,"origin":"","legend":"\u003cp\u003eFigure 4.7: Area wise lernaea parasite prevalence\u003c/p\u003e","description":"","filename":"Picture10.png","url":"https://assets-eu.researchsquare.com/files/rs-4127646/v1/4b602752c2bbb6e0b11658a3.png"},{"id":55101979,"identity":"11761e17-9dae-475e-9191-40ebaa952c4c","added_by":"auto","created_at":"2024-04-22 15:35:57","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":765775,"visible":true,"origin":"","legend":"\u003cp\u003eNile Tilapia fish (Oreochromis niloticus)\u003c/p\u003e","description":"","filename":"Picture3.png","url":"https://assets-eu.researchsquare.com/files/rs-4127646/v1/0aba735f85f0dc2d68c74364.png"},{"id":86568972,"identity":"a1803ee6-3f95-4764-9986-f73efd2f871d","added_by":"auto","created_at":"2025-07-12 13:11:50","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4840016,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4127646/v1/76567295-abf1-4877-aa05-2131c0fc7962.pdf"}],"financialInterests":"","formattedTitle":"Identification of Lernaea Species in Icthyofauna in District Mardan KPK, Pakistan","fulltext":[{"header":"BACKGROUND","content":"\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eA variety of problems is caused by parasites in larger organisms which may lie inside or outside of the body during its life cycle (Llewellyn et al., \u003cspan class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eThe major groups of ectoparasites are Arthropods (crustaceans), Platyhelminthes (monogenesis), and Protozoans. The protozoans are mostly commensal on fish bodies and free living. Most of it causes harmful diseases and is lethal for the host. Ciliates are one of the most common ectoparasites (Buchmann et al., \u003cspan class=\"CitationRef\"\u003e2001\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eLernaea are the copepod parasite which is known as an Anchor worm which may cause infestation and death in a variety of freshwater fish fauna. It may infect the wild species as well as the pond fish. The favorable conditions for the infestation of Lernaea species are the summer season and slow-moving water (stagnant water body). About 110 (hundred and ten) species of the parasite (Lernaea or similar to Lernaea) have been explored up to now. The \u003cem\u003eLernaea cyprinacea\u003c/em\u003e are members of Lernaeid which are found throughout the world. The parasite is common in Cyprinids goldfish, grass carp, and common carp. It can infect the catfish which are mostly stocked in ponds with bighead carp. The Lernaea is the crustacean and copepod that also infects shrimp, crabs, and lobsters in various stages of its life cycles, unlike other free-living copepod species which may not cause disease in the host. The parasites of this group have a complex life cycle, the individual of the group is not passed by the help of an intermediate host organism; whereas it can spread directly from one fish body to the other. The organism needs only a single fish (a member of the class amphibian) to develop from an egg to an adult. The total life cycle of the parasite is completed in 18 to 25 days (Robert \u003cem\u003eet al\u003c/em\u003e., 2001).\u003c/p\u003e\n\u003cp\u003eThe host is the fish which are infected by the parasites. Some parasites have suckers for attachment to the host body while the rest of them lay eggs. It can modify its immunity and biochemistry according to the situation, and cannot be killed or digested. The economic losses occur in the Fisheries sector of Pakistan due to infection of Lernaea species, monogenic species, Argulus species, and Dactylogyrus species (Daulat \u003cem\u003eet al\u003c/em\u003e., 1976).\u003c/p\u003e\n\u003cp\u003eProductivity and fish health are affected by parasites among the ectoparasites. The fish migration and behavior patterns are affected by parasitism. The community structure, survival value, and reproduction of the host are negatively affected by the parasites (Luque and Poulin \u003cspan class=\"CitationRef\"\u003e2008\u003c/span\u003e). Some of the parasites become pathogenic and lethal for fingerlings and fry while most of them show commensalism (Demopoulos \u003cem\u003eet al\u003c/em\u003e., 2015).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eLernea Cyperaceae\u003c/em\u003e Linnaeus is an introduced fish parasite that can cause mortality in various habitats including fish hatcheries and ponds, the species can mostly affect high-density areas with higher temperatures (zman and Rayner 1958). The parasite Laernaea cyprineace is most abundant and harmful among the members of the family Larnaea. (Bilal et al., \u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e). The Lernaeid are the fish parasites that belong to the main group of Crustacea (Copepoda). An extensive morphological fitness (adaptation) is shown by the Lernnaeidae family of cyclopoids which keeps them a little bit distinct from the members of the genus Cyclops and others (Ozel \u003cem\u003eet al\u003c/em\u003e .,2014).\u003c/p\u003e\n\u003cp\u003eThe parasite was first found in Asia later it may be shifted to another continent of the world with farm fish (Innal and Avenant, 2012). The parasite is recorded from various regions of the world including Asia, Southern Africa, Australia, Europe, and its adjacent areas (Hoffman, \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e). The \u003cem\u003eLernaea cyprinacea\u003c/em\u003e is found in Nurseries, brood stocks water hatcheries, and fish ponds of Jessore regions. The parasite (\u003cem\u003eLernaea cyprinacea\u003c/em\u003e) may infect perches, catfish, Minor barbs, and major carps (both Chinese and Indian carps) (Hossain et al., \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eA diverse habitat is occupied by individuals of the family Lernaeidae, parasites are found in marine and freshwater. The family is represented by fourteen (14) genera with one hundred and ten (110) species (Ho, \u003cspan class=\"CitationRef\"\u003e1998\u003c/span\u003e). The parasites are more in number in the aquatic ecosystem and survive with their host in a dynamic equilibrium (Marcogliese, \u003cem\u003eet al\u003c/em\u003e., 2011).\u003c/p\u003e\n\u003cp\u003eHuman health is directly or indirectly affected by infectious. The allergic reaction is caused by either pathogen or living parasites present in the tissues of fish bodies (Buchmann et al., \u003cspan class=\"CitationRef\"\u003e2001\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eThe adult member of Lernaea cyprinacea attached to the skins and muscles of the host leaves an opening to enter the secondary microbe and cause death to the infected fish (Van Duijn \u003cem\u003eet al\u003c/em\u003e., 1973).\u003c/p\u003e\n\u003cp\u003eThe pathogenicity of lernaeids largely depends on the size of their host and attachment site preferences. Aggregated infections of the mouth and lips occur in Barbus altianalis with Lernaea barinmiana and in cichlids by Lernaea cyprinacea, fin infections caused by Lernaea lophiara in cichlids (Fryer, \u003cspan class=\"CitationRef\"\u003e1968\u003c/span\u003e; Paperna, \u003cspan class=\"CitationRef\"\u003e1996\u003c/span\u003e). Fish with large numbers of parasites will be thin and their body color becomes darkened, also extensive abrasions appear with loosing and sloughing on its skin. Severe destruction of the gills occurs as well as losses and mortalities up to 50% (Eissa \u003cem\u003eet al\u003c/em\u003e., 2004).\u003c/p\u003e\n\u003cp\u003eThe life cycle of the parasite Lernaea has nine stages, out of which three stages are free-living naupliar, five copepodid, and one stage of an adult. During the fourth stage, copulation of both adult males and females occurs at the first host. When the copulation insures the male is dyed whereas the female metamorphoses and enters the anterior tissues of the host body, laying eggs (Nagasawa et al., \u003cspan class=\"CitationRef\"\u003e2007\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eA tremendous modification in cephalothorax has been shown by the adult female of the genus Lernaea. The body is vermiform and elongated, and the mouth of the parasite female is much reduced. The head is changed into 4 horn shaped compartments which are now cylindrical and long, the anterior appendages are simple while the two posterior ones are bifurcated. The parasites use the horns for attachment to the host body and it’s buried under the epidermis. The abdomen is bent toward the dorsal and short in length. The egg sacs protrude beyond the body while the genital pore is present near the posterior end of the body. The immature forms of both sexes (male and females) and an adult male are free-living; whereas the mature or an adult female of learned have the capability of parasitism. \u003cem\u003eLernaea cyprinacea\u003c/em\u003e attacked massively with the highest mortality and pathogenicity, mostly in the season of summer. The parasite caused ulceration and hemorrhages along with penetration (Carnevia \u003cem\u003eet al\u003c/em\u003e., 2003).\u003c/p\u003e\n\u003cp\u003eThe native as well as introduced species of fish fauna in the South American continent are parasitized by 3 species of Lernea, (Boxshall et al., \u003cspan class=\"CitationRef\"\u003e1997\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eThe parasite was recorded in the fish fauna of Argentina, Brazil, Colombia, Chile, and Uruguay (Habit \u003cem\u003eet al\u003c/em\u003e., 2015).\u003c/p\u003e\n\u003cp\u003eThe harmful effects of the concerned parasite were observed both in wild and cultured fish populations (Yanong et al., \u003cspan class=\"CitationRef\"\u003e2002\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eThe species causes serious economic loss in various countries of the world (Rogers, \u003cspan class=\"CitationRef\"\u003e1979\u003c/span\u003e). Lernaeosis is a major disease that is caused by the parasite in the hosts (Hua et al., \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eThe parasites may cause huge destruction and dangerous diseases in fish fauna, may lead to the extinction of species, and can damage the health of humans by consuming them as food (Habib et al., \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eEconomic losses have been observed in various formed fishes due to lernaeid ectoparasites globally (Kir \u003cem\u003eet al\u003c/em\u003e., 2007).\u003c/p\u003e\n\u003cp\u003eLernaeid copepods are considered one of the harmful parasites of freshwater ecosystems; a few individuals can kill young fish (Kabata \u003cem\u003eet al\u003c/em\u003e., 1987).\u003c/p\u003e\n\u003c/div\u003e\n\n"},{"header":"AIMS AND OBJECTIVES","content":"\u003col\u003e\n\u003cli\u003e\n\u003cp\u003eTo find the prevalence of Lernaea species in the study area.\u003c/p\u003e\n\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n\u003ch2\u003eStudy Area\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eThe study was conducted in District Mardan, river Kalpani, ponds of Tehsil Takht Bhai and\u003c/p\u003e\n\u003cp\u003eShergarh Khyber Pakhtunkhwa Pakistan. District Mardan lies between 34.1986˚ N, latitude and\u003c/p\u003e\n\u003cp\u003e72.0404˚ E longitude with a total area of 1632 km\u003csup\u003e2\u003c/sup\u003e. Tehsil Takht Bhai lies between 34.3314˚ N, Latitude, and 71.9046˚ E, longitude with a total area of 353.0 km\u003csup\u003e2\u003c/sup\u003e. Shergarh is a city of Tehsil Takht Bhai which lies between 34.39349˚ N latitude and 71.8996˚ E longitude.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eFigure 3.1: Study area and data collection center for Mardan District (figures were altered using Google.com)\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n\u003ch2\u003eFish sample collection\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eBetween February to August 2023, fish samples were collected from 9 ponds of District Mardan, Tehsil Takht Bhai, Shergarh, and river Kalpani Khyber Pakhtunkhwa Pakistan.\u003c/p\u003e\n\u003cp\u003eFish samples were tallied to determine whether they were positive or negative for Lernaea infection. The number of Lernaea on each fish was counted in positive samples to evaluate the degree of infestation.\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n\u003ch2\u003eFish sample processing\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eThe collected fish samples from the study site were brought immediately in oxygenated bags to the lab, department of Zoology GDC Lundkhwar. Morphometric measurement was done by taking total length [TL] fork length [FL] standard length [SL] anal length [AL] pre orbital length [POL] post orbital length [POL] head length [HL] and body depth [BD]. The fish were weighed by using a digital scale and species identification was done by using the method of Hoffman, \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n\u003ch2\u003ePreliminary Treatment of Fish\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eThe fishes were first given serial numbers, and then the length [in cm] of the fishes was measured from the tip of the snout to the end of the caudal fin using a measuring scale. The weight was taken by using a digital weighing scale. The external body surface and the structures, notably, the skin and fins were thoroughly examined for the presence of lernaea parasites.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n\u003ch2\u003eSkin\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eThe scales of the scaly fishes were removed softly with the help of a scalpel. They were cleaned thoroughly with running water and then with distilled water to ascertain the presence of lernaea ectoparasite. Mucus and epithelial tissue are also gently scraped wherever necessary. The materials were then transferred to a small drop of water on a glass slide and covered with a small size cover glass. The materials were then examined under a microscope and recorded.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n\u003ch2\u003eFins\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eThe fins were examined under the microscope after they were cut off and transferred to a Petridis with water. Epithelial scrapings from the fins were made and subjected to microscopic examination.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n\u003ch2\u003eParasitological Study\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eFor parasite collection and identification, the fish samples were deeply examined with naked eyes after that they were examined with a hand lens; naked eye parasites including lernaea species were removed by using forceps and preserved in 70% ethanol. For microscopic ectoparasite, examination was conducted using the method of scraping (Kabata, 1985). For further studies, the isolated parasites were collected, counted, and processed following Gussev (1986).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec10\" class=\"Section3\"\u003e\n\u003ch2\u003eMorphological Identification\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eLernaea was taken from the fish body using tweezers, rinsed with physiological NaCl, and preserved with lacto phenol. The Lernaea samples used for identification were preserved in absolute ethanol. Lernaea was observed in the holdfast, abdomen, position, and number of legs. The body and anchor length were measured. Lernaea was identified using Kabata\u0026rsquo;s [15] identification key.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003c/div\u003e"},{"header":"RESULTS","content":"\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eThis research aimed to assess the prevalence, intensity, and abundance of Lernaea species within the aquaculture population in the district of Mardan, Pakistan. Over a period of six months, from February to August 2023, a total of 159 fish samples were collected. These samples were sourced from both the Kalpani River and a hatchery in Shankar Mardan. The examination for lernaeid parasites was conducted in the laboratory of the Department of Zoology at GDC Lundkhwar.\u003c/p\u003e\n\u003cp\u003eThe overall prevalence of Lernaea species disease in Dist. Mardan aquaculture was found as 64.77%.\u003c/p\u003e\n\u003cp\u003eThe prevalence of Lernaea species disease in where much positive in the selected samples summarized in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4.1\u003c/span\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 4.1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eShows the total prevalence along with positive and Negative samples\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSr.\u003c/p\u003e\n\u003cp\u003eNo\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eLength\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eTotal samples\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePositive\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eNegative\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePositive\u003c/p\u003e\n\u003cp\u003e%age\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eNegative\u003c/p\u003e\n\u003cp\u003e%age\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eTotal P-Value\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10-60cm\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e114\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e74\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e40\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e64.91\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e35.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e64.77%\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e81-90cm\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e29\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e16\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e64.44\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e35.55\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eA total of 114 samples were collected from fish ranging from 10 to 60 cm in length. Among these, 74 samples tested positive, indicating a frequency of 64.91%. Additionally, 45 samples were obtained from fish measuring 81 to 90 cm in length, with 29 of them showing positive results, resulting in a prevalence of 64.44%. The above values are graphically explained in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4.1\u003c/span\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eFor fish within the 10\u0026ndash;20 cm size range, a maximum of 29 samples were collected, with 15 of them yielding positive results, indicating a positivity rate of 51.72%. In the 21\u0026ndash;40 cm size category, 35 samples were obtained, and 28 of them tested positive, resulting in a higher positivity rate of 80%. Among the 41\u0026ndash;60 cm size group, 50 samples were collected, with 31 showing positive results, yielding a positivity rate of 62%. Lastly, in the 81\u0026ndash;90 cm size range, 45 samples were taken, and 29 of them tested positive, indicating a prevalence value of 64.44%.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab3\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 4.2\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eLength-wise distribution of samples and their disease prevalence value\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSr.\u003c/p\u003e\n\u003cp\u003eNo\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eLength\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eTotal Samples\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePositive\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eNegative\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePrevalence value\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10\u0026ndash;20 cm\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e29\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e51.72\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e21\u0026ndash;40 cm\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e35\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e28\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e41\u0026ndash;60 cm\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e31\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e19\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e62\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e81\u0026ndash;90 cm\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e29\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e16\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e64.44\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eThe distribution of Lernaea cyprinacea on various parts of Nile Tilapia (Oreochromis niloticus) is illustrated in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4.4\u003c/span\u003e. The figure reveals that the area around the base of the dorsal fin exhibited the highest infestation by these copepods, with a recorded presence of 78 instances of Lernaea cyprinacea.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eA collection of 159 specimens of freshwater fish fauna was gathered and identified, encompassing seven distinct species. Within the studied area, a total of five parasite species were observed in association with these fish. The parasite species were \u003cem\u003eCarpora arthritis\u003c/em\u003e, \u003cem\u003eSaprolegnia\u003c/em\u003e, \u003cem\u003eSpherosporaauretus\u003c/em\u003e, \u003cem\u003eLernaeacyprinacea\u003c/em\u003e, and \u003cem\u003eMicrosporadia\u003c/em\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab4\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 4.3\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eParasite species distribution\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eName\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eCarpora arthritis\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eSaprolegnia\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eSpherosporaauretus\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eLernaeacyprinacea\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eMicrosporadia\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eNo of species\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e25\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e27\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eWe utilized a categorization system that classifies infestation levels as light (1\u0026ndash;5 Lernaea per fish), moderate (6\u0026ndash;10 Lernaea per fish), or heavy (more than 10 Lernaea per fish).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab5\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 4.4\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eLernaea per fish degree of infestation.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSr.\u003c/p\u003e\n\u003cp\u003eNo\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eLernaea per fish\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eNo of infected sample\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003epercentage\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u0026ndash;5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e32\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e20.12\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6\u0026ndash;10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e52\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e32.70\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003emore than 10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e75\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e47.16\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eA total of 159 fish samples were gathered from various locations within District Mardan. Specifically, 45 samples were collected in Takht Bhai, 49 samples in Shergarh, and 65 samples were obtained from River Kalpani, as detailed in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4.5\u003c/span\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab6\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 4.5\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eArea wise lernaea parasite prevalence\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSr. no\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eLocation\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eNumber of\u003c/p\u003e\n\u003cp\u003eSamples\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePositive\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePositive Percentage\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTakhtbhai\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e32\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e71.11\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eShergarh\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e49\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e17\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e34.69\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eRiver Kalpani\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e65\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e35.38\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eAquaculture stands out as one of the globe's most economically significant sectors, and fish, being a crucial nutritional resource, holds immense value for humanity. The socio-economic development aimed at fostering healthy ecosystems and ensuring the survival of life on Earth necessitates a concerted effort to safeguard water resources and the ecosystems connected to them.\u003c/p\u003e\n\u003cp\u003eThe study conducted from February to August 2023 aimed to evaluate the prevalence and infestation levels of Lernaea parasites in fish. Sample collection took place in different ponds near the Takhtbhai archaeological site, spanning District Mardan, Tehsil Takhtbhai, Shergarh, and the Kalpani River in Khyber-Pakhtunkhwa, Pakistan.\u003c/p\u003e\n\u003cp\u003eA total of 159 specimens of freshwater fish fauna were collected and identified, representing seven distinct species. Within the specified area, a total of five parasite species were identified in association with these fish. The parasite species were \u003cem\u003eCarpora arthritis\u003c/em\u003e, \u003cem\u003eSaprolegnia\u003c/em\u003e, \u003cem\u003eSpherosporaauretus\u003c/em\u003e, \u003cem\u003eLernaeacyprinacea\u003c/em\u003e and \u003cem\u003eMicrosporadia\u003c/em\u003e. The \u003cem\u003esaprolignia\u003c/em\u003e was more frequent than the rest of four parasites. The parasites infect the whole body of the host including the gills, head and skin. The all entire system of some hosts was badly infected by the parasites. Khan et al., (\u003cspan class=\"CitationRef\"\u003e2023\u003c/span\u003e) collected Seventy-six (76) specimens of fish fauna were collected from seventeen (17) ponds during the period from September to November 2021. The sampling locations encompassed various districts, including Swabi, Mardan, Charsadda, and Peshawar in the Khyber-Pakhtunkhwa province. The aim of the study was to investigate the ectoparasites associated with the fish fauna in these regions.\u003c/p\u003e\n\u003cp\u003eThe outcomes of our study unveil a significant prevalence of Lernaea infestation within the fish populations in our designated study area. Following the categorization method introduced by Kriswijayanti (2019), our assessment differentiated infestations into three levels: light (1–5 Lernaea per fish), moderate (6–10 Lernaea per fish), and heavy (more than 10 Lernaea per fish).\u003c/p\u003e\n\u003cp\u003eThe findings indicate varying degrees of susceptibility to Lernaea infestation among the fish in the Takhtbhai region. Our observations align with previous research by Khan et al., (\u003cspan class=\"CitationRef\"\u003e2023\u003c/span\u003e), who reported a similar prevalence of Lernaea in fish populations in neighboring regions.\u003c/p\u003e\n\u003cp\u003eOur study highlights the crucial need to comprehend the presence and impact of Lernaea parasites to facilitate effective conservation endeavors. Notably, it emphasizes the necessity for targeted mitigation strategies, especially for fish populations experiencing heavy infestations. This emphasis on tailored strategies aligns with the conservation recommendations put forth by Ahmed and Rahman (2017) in their research on parasite management within local aquatic ecosystems.\u003c/p\u003e\n\u003cp\u003eAbout forty three (43) species of fishes and three (species) of amphibians were identified up to species level in the area which were in infected by \u003cem\u003eLerneacyprinacea\u003c/em\u003e. Charciform was more parasitized among the rest of seven (7) orders in the continent. Ahnelt et al., (\u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e) reported that a variety of fish fauna is infected by the parasitic copepod \u003cem\u003eLernaeacyprinacea\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003eEctoparasite infestation stands out as a primary contributor to mortality among infected fish species. The prevalence of the Lernaea cyprinacea parasite is notably higher in pelagic species compared to smaller benthic fish species. Tayyab (2017) conducted a study on the River Indus, aiming to investigate parasite infestation within the freshwater ecosystem of the specified study area.\u003c/p\u003e\n\u003cp\u003eThe prevalence of Lernaea species was found to be significant, accounting for 39.47% of the observed parasites in the fish fauna. This species demonstrated higher lethality compared to other parasite species. Following Lernaea, Gyrodactylus and Dactylogyrus species constituted 19.73%, Icthyopthyrius multifilis species accounted for 17.10%, thelohanellus species represented 11.84%, Loxophyllum species constituted 10.52%, and Piscinoodinium pillulare species made up 7.89% of the observed parasites.\u003c/p\u003e\n\u003cp\u003eAll seven species collected during this study were identified for the first time in the study area. According to Innal \u003cem\u003eet al\u003c/em\u003e., (2020), \u003cem\u003eLernaea cyprinacea\u003c/em\u003e, commonly known as anchor worm, is prevalent in a wide range of hosts and belongs to the genus Lernaea. This species has been documented in various studies in the Mediterranean region, specifically in Turkey. The region encompasses a diverse group of cypriniforms, and the parasites were identified in both native and introduced fish populations within the area.\u003c/p\u003e\n\u003c/div\u003e\n"},{"header":"FINDINGS","content":"\u003cp\u003eThis investigation was conducted in Takhtbhai area, Khyber-pakhtunkhwa, Pakistan, from Feb. 2023-July 2023. The purpose was to know the number of Lernaean parasites living and the level of infestation in freshwater fish. One hundred and fifty-nine specimens belonging to twelve members of seven species of fish from different ponds and the Kalpani River were collected. The result of this research, Trematoda found 5 species of which Saprolegnia were the most abundant. Infected juveniles were observed at higher and lower levels, and the study employed a categorization method, where the degree of severity was noted to determine the difference in the susceptibility of fish to Lernaea infestation. The outcome correlate with past findings meaning highlighting how necessary the required counter-attack techniques and the great conservation solutions is, especially in those fish heavily infected by Lernaea. This research has also been valuable because it has demonstrated the richness of fish and amphibian species in the area, the fact that all 43 of the fish species are infected by Lernaea cyprinacea. A study found that Lernaea infestation in addition to other external parasites was the main factor leading to the fish mortality. Lernaea cyprinacea was revealed as the most commonly occurring parasite whereas in comparison the size of the other organisms was smaller. This research highlights how parasitic infestations of freshwater ecosystems may affect the ecologic dynamics, and the parasitic infestations should be considered in management plan in the region to conserve the system effectively\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eAs a conclusion, this research analysis for district Mardan region discourages the intensive infection of the louse fish parasite within the freshwater fishes and underlines that applicable unsustainable measures are key to remedying the infestation varying from low to grave level. The conclusions provide the evidence of high vulnerability of these fish populations to Lernaea infestation and can be compared with previous studies, negative implication can be made that appropriate conservation measures should be adopted. The data collected through this pilot investigation regarding the different types of fish and amphibians that suffer Lernaea cyprinacea infestation are important to the greater ecological picture on how ectoparasites in freshwater can affect the balance. The summarized study facilitates the accumulation of new helpful information with formulation of conservation strategies that promote the wellbeing and sustainability of water bodies in the Khyber-Pakhtunkhwa province, Pakistan.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e"},{"header":"Abbreviations","content":"\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"585\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"37.11340206185567%\" valign=\"top\"\u003e\n \u003cp\u003eKPK \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.371134020618557%\" valign=\"top\"\u003e\n \u003cp\u003e:\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50.51546391752577%\" valign=\"top\"\u003e\n \u003cp\u003eKhyber Pakhtunkhwa \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"37.11340206185567%\"\u003e\n \u003cp\u003eGDC \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.371134020618557%\"\u003e\n \u003cp\u003e:\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGovt Degree college\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"37.11340206185567%\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;TL \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.371134020618557%\"\u003e\n \u003cp\u003e:\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50.51546391752577%\"\u003e\n \u003cp\u003eTotal length\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"37.11340206185567%\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;FL \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.371134020618557%\"\u003e\n \u003cp\u003e:\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50.51546391752577%\"\u003e\n \u003cp\u003eFork length\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"37.11340206185567%\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;SL \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.371134020618557%\"\u003e\n \u003cp\u003e:\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50.51546391752577%\"\u003e\n \u003cp\u003eStandard length\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"37.11340206185567%\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;AL \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.371134020618557%\"\u003e\n \u003cp\u003e:\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50.51546391752577%\"\u003e\n \u003cp\u003eAnal length\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"37.11340206185567%\"\u003e\n \u003cp\u003ePOL \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.371134020618557%\"\u003e\n \u003cp\u003e:\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50.51546391752577%\"\u003e\n \u003cp\u003ePre orbital length\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"37.11340206185567%\"\u003e\n \u003cp\u003eHL \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.371134020618557%\"\u003e\n \u003cp\u003e:\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50.51546391752577%\"\u003e\n \u003cp\u003eHead length \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"37.11340206185567%\"\u003e\n \u003cp\u003eBD\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.371134020618557%\" valign=\"top\"\u003e\n \u003cp\u003e:\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50.51546391752577%\" valign=\"top\"\u003e\n \u003cp\u003eBody depth\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Declarations","content":"\u003cp\u003ethe Ethical Review Committee (ERC) of Abdul Wali Khan University Mardan has granted ethical approval for this study.\u003c/p\u003e\n"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAhnelt H, Konecny R, Gabriel A, Bauer A, Pompei L, Lorenzoni M, Sattmann H (2018) First report of the parasitic copepod Lernaea cyprinacea (Copepoda: Lernaeidae) on gobioid fishes (Teleostei: Gobonellidae) in southern Europe. 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Co-invasion of anchor worms Lernaea cyprinacea\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e(Copepoda: Lernaeidae) in some freshwater fishes of the Kor River Basin, Southwest of Iran with some remarks on the ecological aspects of lernaeosis in the country. Iranian Journal of Fisheries Sciences 15(1) 369\u0026ndash;389\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSilva-Souza A, Almeida T, Machado S (2000) Effect of the infestation by Lernaea cyprinacea Linnaeus, 1758 (Copepoda, Lernaeidae) on the leucocites of Schizodon intermedius Garavello and Britski, 1990 (Osteichthyes, Anostomidae). Revista Brasileira de Biolog\u0026iacute;a 60(2):217\u0026ndash;220\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTayyab M, Gul S, Rehman UH, Rehman UA, Saeed K, Khan S, Naz N, Ahmad A (2017) J Entomol Zool Stud 5(4):422\u0026ndash;426\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTekin-\u0026Ouml;zan, Kir SI, Barlas M (2008) Helminth parasites of common carp. Cyprinus carrpio\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eL., 1758) in Beyșehir lake and population dynamics related to month and host size. Turk\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFish J Aquat Sci, 8: 201\u0026ndash;205\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWaicheim AM, Arbetman M, Rauque C, Viozzi G (2019) The invasive parasitic copepod Lernaea cyprinacea: updated host-list and distribution, molecular identification and infection rates in Patagonia. Aquat Invasions 14(2):350\u0026ndash;364\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWoo PTK (2002) Fish Diseases and Disorders. CABI International Wallingford Oxfordshire OX10 8DE, UK. 808 pp\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWoo PTK (1995) Fish diseases and disorders. CAB, Int. Wallingford, Oxon, Uk\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVan Duijn J (1973) Diseases of fishes Illffe books London\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYanong RP, Francis-Floyd R, Curtis E, Klinger RE, Cichra MF, Berzins (2002) I. K.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlgal dermatitis in cichlids Journal of the American Veterinary\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKriswijayanati BD, Kismiyati K, Kusnoto K (2019) Identifikasi dan derajat infestasi\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLernaea pada ikan maskoki (Carassius auratus) di kabupaten Tulungagung, Jawa Timur (Identification and degrees of Lernaea infestation in Goldfish (Carassius auratus) at Tulungagung, East Jawa). J Aquac Fish health, 3(1): 1\u0026ndash;7\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Lernaea, hatchery, Mardan, Pakistan, Prevalence, Kalpani, Parasite, and Fish","lastPublishedDoi":"10.21203/rs.3.rs-4127646/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4127646/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eLernaea cyprinacea, a parasitic cyclopoid copepod, is found to parasitize in the brood stock, nursery and culture system in the freshwater hatchery of Mardan regions. The fish were severely infected on their body surface showed dull and sluggish movements. This research, carried out in the district Mardan Khyber-Pakhtunkhwa, Pakistan, spanning from February to August 2023, delves into the prevalence and infestation dynamics of Lernaea parasites in freshwater fish in river Kalpani. Through a comprehensive analysis of 159 specimens collected from various aquatic sources, the study identifies the presence of five parasite species, notably with Saprolegnia as the predominant one. Utilizing a categorization method, the investigation discerns varying degrees of infestation, underscoring the susceptibility of local fish populations to Lernaea. These findings corroborate with earlier research, accentuating the pressing need for targeted mitigation strategies and robust conservation initiatives. Additionally, the study unravels the profound impact of \u003cem\u003eLernaea cyprinacea\u003c/em\u003e on 103 fish, emphasizing its pivotal role as a primary cause of fish mortality. This research not only contributes crucial insights for shaping effective conservation measures but also proves imperative for sustaining the overall health and biodiversity of freshwater ecosystems in the region.\u003c/p\u003e","manuscriptTitle":"Identification of Lernaea Species in Icthyofauna in District Mardan KPK, Pakistan","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-22 15:35:52","doi":"10.21203/rs.3.rs-4127646/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"8a1e138d-b679-4a50-b37b-531f0564c01c","owner":[],"postedDate":"April 22nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-07-12T13:03:40+00:00","versionOfRecord":[],"versionCreatedAt":"2024-04-22 15:35:52","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4127646","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4127646","identity":"rs-4127646","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}