Spectrum of Zoonotic Fish Parasites in Lahore, Pakistan: An Anatomopathological analysis for tissue damage

doi:10.21203/rs.3.rs-4816928/v1

Download PDF

Research Article

Spectrum of Zoonotic Fish Parasites in Lahore, Pakistan: An Anatomopathological analysis for tissue damage

https://doi.org/10.21203/rs.3.rs-4816928/v1

This work is licensed under a CC BY 4.0 License

You are reading this latest preprint version

Fish parasites are a leading cause of fatality as well as a threat to public health and the economy as well. Different species of parasites are reported in several types of fish. To protect the farm from parasitic infestations, and mitigate the threat to public health, regular surveillance in the fish farms should be performed. Therefore, 100 random samples from the fisheries complex Manawa, Lahore were taken from February 2019 to July 2019 to determine varieties of fish parasites, and for the observation of histopathological changes in fish. The study reveals a 3% prevalence of the fish parasites in the region of surveillance, and two species Anisakis simplex and Lernaea cyprinacea are found in the samples. Moreover, the histopathological changes including liver, skin, gills, and intestines of positive fish samples are observed.

Fish

Parasites

Prevalence

Histopathology

Parasitic infestations not only cause fatality in fish and play a crucial role in economic losses but their zoonotic importance cannot be neglected due to the use of raw, steamed, and semi-cooked fish. According to research, more than 18 million people were infected with fish-borne zoonotic trematodes in 2002 worldwide. In the present study, two zoonotic parasite species were isolated from fish farms that had not been previously reported in this region. Therefore, it is very important to disseminate the outcomes of this novel finding to the research community and the general public. Moreover, the histopathological changes caused by these parasites in fish were studied. As there are only a few studies on fish pathology in Pakistan therefore the novelty of this research must be promulgated.

Anisakis simplex is a nematode parasite that is found all over the world, and its life cycle is characterized by numerous stages in which it parasitizes sea animal species. Third-stage larvae are found in the celomic cavity and viscera of teleost fish and cephalopods. The larvae are 2–4 cm in length and white (Bao et al., 2019). It causes a foodborne zoonotic disease, called anisakiasis. It is a gastrointestinal tract infection of a human being caused by using raw or undercooked seafood containing larvae of A. simplex. It has a life cycle including marine mammals and fish. Humans are considered accidental hosts of this parasite (Golden et al., 2023). Thiel reported 11 cases of larval anisakiasis in human beings with third-stage larvae The most important intermediary host species are squid, mackerel, red mullet, sardine, and salmon. The acute form is caused by third-stage larvae, which are called intestinal or gastric anisakiasis. In this type of infection, the larvae can be found in the mucosa or submucosa after penetration of the host stomach or intestinal wall. It is characterized by nausea, vomiting, and epigastric pain. Chronic anisakiasis is characterized by eosinophilic granulomas, abdominal pain, and eosinophilia diarrhea (Adroher-Auroux and Benítez-Rodríguez, 2020).

To prevent anisakiasis infection, the most effective treatment is taken by the Commission Regulation. Some measures are also taken to avoid the consumption of raw or undercooked fish to inactivate the larvae. These treatments consist of freezing fish at − 35°C for at least 15 hours or − 20°C for at least 24 hours. Another way to inactivate the larvae is by cooking fish at an internal temperature of 65°C for at least 30 sec or at 60°C for at least 1 minute (Debenedetti et al., 2019)

Lernaea cyprinacea is a Maxillopoda parasite that belongs to the genus Lernaea that has a huge impact on the economy due to its various epizootics occurrence in different countries of the world (Raja et al., 2023). It is the most common and opportunistic ectoparasites which appears throughout the world in freshwater fish and cause a disease called lernaeosis. It has been isolated from 11 native fish of Cyprinids and Cichlidae and has been observed on the gills and skin of freshwater fish (Chakona et al., 2019). Adult females of L. cyprinacea attach to body surfaces of fish involving the head, caudal, ventral, and dorsal regions, but commonly to the bases of fins (Rahmati-Holasoo et al., 2023). It is a thermophilic organism commonly occurring in the summer season with temperatures ranging from 25 to 28°C. Due to its thermophilic ability, it grows faster in warm environments than colder (Jerônimo et al., 2022). It generally occurs in cyprinid fish feeding on the host's tissues and blood (Nur et al., 2022).

It is often called anchor worm and is a parasite of freshwater fish in different regions throughout the world This species commonly occurs in goldfish (Carrassius auratus), Crucian carp (Carrassius carrassius), and common carp fish (Cyprinus carpio) respectively. This parasite is distributed naturally in central Asia, the southern region of Siberia, and Africa, and has also been reported in Israel, Japan, and Europe (Trilles et al., 2011).

Therefore, this research aimed to determine various varieties of fish parasites and to assess the histopathology of the liver, intestine, skin, and gills of infected fish with parasites in Fisheries Complex Manawa, Lahore Pakistan.

Experimental design:

The present research was done for a period of 6 months from February 2019 to July 2019. Since there was no prior study conducted in this region. A total of (n = 100) live fish samples were collected on a random basis from Fisheries Complex Manawa Lahore, Pakistan, and placed in an aquarium containing freshwater with aerators and brought to the Department of Pathology at the University of Veterinary and Animal Sciences, Lahore for the examination of parasites. The total length (standard length) and weight of each fish were calculated and specimens were sexed by observation of gonads and anal fin. They were washed carefully with tap water and were preserved in 10% formalin which was examined for the occurrence of parasites. Each preserved fish specimen before dissection was subjected to tap water washed thoroughly to remove any killing or fixatives and desiccated using a paper towel.

Dissection procedure:

External parasites were examined on the outer surface of the fish and taken out by forceps. For examination of internal parasites, fish were opened dorso-ventrally and examined. The Gills, body cavity, liver, and gastrointestinal tract of fish were removed for examination of parasites, which were washed in a Petri dish containing tap water. The GIT was cut into small pieces e.g. small intestine, large intestine, and stomach. A longitudinal incision was made in the intestine and the intestinal contents were transferred to another Petri dish for finding parasites. A few drops of methanol were placed to loosen the attachment of the parasite and carefully transferred to a bottle containing normal saline. The bottle was shaken thoroughly, which loosened the attachment of the parasite to the gills and skin. The parasite was taken out carefully using force and placed in separate vials.

Parasites identification:

For the identification of parasites, an Aceto-Carmine stain was used. Each preserved parasite was first rehydrated (washed) with tap water and then dehydrated by using 70%, 50%, and 30% alcohol each for 5 minutes. A drop of Aceto-Carmine was put in for 2 minutes and the specimen was washed through using tap water. The parasite was mounted in Canada balsam and a coverslip was kept on it and a permanent slide was made. Now the parasite was ready for identification and kept under a light microscope (100X). All the pictures were taken by using a camera and morphologically identified by the key (Hua et al., 2019);(Innal and Avenant-Oldewage, 2012).

Prevalence rate determination:

The prevalence rate was determined by the following formula as described by (Justus et al., 2012).

Prevalence rate (%) = Number of hosts infected

---------------------------------- × 100

Total number of hosts examined

Histopathological examination:

Tissue samples were collected from fish skin, gills, liver, and intestine for histopathological purposes, that was transferred to formalin jars, which contain 15 times 10% buffered formalin solution, embedded in paraffin wax, sectioned and stained with hematoxylin and eosin (H & E) as followed by (Slaoui and Fiette, 2011). Finally, the prepared slides were examined under a light microscope.

Sample size calculation:

The sample size was calculated as 97 ~ 100 using the formula previously described by (Thrusfield, 2007). \(\:Sample\:size\:\left(n\right)=\frac{{Z}^{2}pq}{{d}^{2}}\) Where Z² = 1.645² at 90% confidence level, p is expected prevalence (10%), q = 1-p (90%), and d is desired absolute precision (5%).

Statistical analysis:

Excel 2016 software was used to process the initial data and ensure the entries' precision. A Fisher exact test was performed to determine the difference among the proportions of the variables. The adjusted odds ratio was calculated using the binary logistic regression model. The results were displayed as percentages (%) expressed in 95% confidence intervals (95%CI). Histopathological scoring was performed based on the intensity of lesions in the body tissues and the mean rank scores were compared using the Wilcoxon rank-sum test. The analysis used Mlinitab® 21.4 (Mathews, 2004) and GraphPad Prism 9. Results were considered statistically significant if the p-value was less than 0.05.

The histopathological analysis of tissue samples from Nile Tilapia and Grass Carp showed that there were some qualitative differences in the organ damage of both fish species (Table 1). However, the Wilcoxon rank-sum test did not reveal any significant differences in the mean rank scores of the histological changes in various organs of both fish species. Gill damage was observed, with Nile Tilapia having marked necrosis, while there was comparatively higher congestion in the grass carp. The histological changes observed in hepatic tissue were congestion and hydropic degeneration. However, a non-significant difference in these changes was observed between both species. However, higher mean ranks were observed in the histopathological score in intestinal and cutaneous tissue of Nile tipalia. However, no significant difference in the mean ranks could be observed (Fig. 1)

Table 1. Histopathological scores indicating tissue changes in Nile Tilapia and Grass Carp.

Tissue	Histological Change	Nile Tilapia	Grass Carp

Gills	Necrosis	++	+
Gills	Congestion	-	++
Hepatic tissue	Congestion	++	++
Hepatic tissue	Hydropic Degeneration	+	++
Intestinal tissue	Leukocytic infiltration	++	+
Intestinal tissue	Desquamation	+	++
Cutaneous tissue	Epidermal Necrosis	++	++
Cutaneous tissue	Dermal Necrosis	+	++

(-) denotes mean normal, (+) signifies mild change, (++) indicates moderate change, and (+++) represents severe change.

In the present study, the univariate analysis of the demographic variables revealed that 60% of the total sample was Grass carp, while Nile tilapia accounted for 40%. 62% of the total samples were male and 38% were females. Body length and weight distribution of data revealed that 62% of subjects were 10–15 cm ranging from 200 to 250 grams body weight. Whereas, 38% had a body length of more than 15 cm and had more than 300 grams of body weight (Fig. 2).

The results of the study revealed a total of 3% prevalence of parasites. Out of positive individuals, one Nile tilapia was found positive for Anisakis simplex which was observed macroscopically (Fig. 3). Two Grass carp tested positive for Lernaea cyprinacea. Interestingly, none of Nile tilapia and Grass Carp was found positive for Lernaea cyprinacea and Anisakis simplex, respectively (Fig. 4).

Results of the bivariate analysis revealed that 2 individuals (3.34%) among grass carp tested positive for parasites compared with only 1 (2.5%) fish positive among Nile tilapia. The odds ratio for Grass carp being infested was 2.01, suggesting twofold odds of being positive compared to Nile tilapia. Specifically, among females, 1 (2.64%) tested positive, resulting in an odds ratio of 1.06. In comparison, among males, 60 (96.78%, 95% CI [88.98% were negative and 2 (3.23%) were tested positive.

For fish greater than 15 cm in length, 36 individuals (94.74%, 95% CI [82.72% − 99.07%]) were classified as negative and 2 individuals (5.27%, 95% CI [0.94% − 17.29%]) as positive for the outcome, yielding an odds ratio of 4.18 (95% CI [0.30–58.21]). In contrast, among fish measuring 10–15 cm, 61 were negative (98.39%, 95% CI [91.42% − 99.92%]) and only 1 was positive (1.62%, 95% CI [0.09% − 8.59%]).

This research study was performed to know the occurrence of Anisakis simplex and Lernaea cyprinacea parasites in fish as well as to analyze the histopathology of the liver, skin, intestine, and gills of fish that were infected by parasites. In this study, an overall prevalence rate of 06% was noted in Nile tilapia and Grass carp species. It was observed that a significant difference was noted in the prevalence of parasites among fish species at P > 0.05. The prevalence of Lernaea cyprinacea was higher in Grass carp (3.5%) followed by Anisakis simplex in Nile tilapia (2.5%) fish. Our prevalence contradicts the reports of (Koyun et al., 2015) and (Chen and Shih, 2015) noted that the Anisakis spp. larvae showed a prevalence of 39.9% in different fish species and L. cyprinacea showed a prevalence of 5% in Cyprinion macrostomum Heckel fish. In another study done by (Innal and Avenant-Oldewage, 2012), L. cyprinacea showed a prevalence of 15% in mosquitofish. This study compares with the reports given by (Chen and Shih, 2015), that Anisakis larvae showed a high prevalence rate of 72.8% in spotted mackerel Scomber australascius fish. The difference was not statistically significant between sexes, but the prevalence was slightly higher in males (3.4%) than females (2.6%) fish. This study corresponds with the study of (Akinsanya et al., 2008) and (Allumma and Idowu, 2011), regarding sex who found that male fish showed higher prevalence than female fish. This statement agrees with the research of (Innal and Avenant-Oldewage, 2012), who found that male fish were more heavily parasitized than female fish. This finding contradicts the study of (Eissa et al., 2018), who concluded that Anisakis spp. was more prevalent in female fish than male fish. The prevalence of L. cyprinacea and A. simplex parasites was also evaluated based on different length and weight (age) categories. Their prevalence was statistically significant at P < 0.05, which showed that the prevalence was higher in length and weight of fish (> 15 cm, > 300g) as compared to the prevalence of fish (10–15 cm, 200-250g) respectively. It was concluded that a higher parasite infestation occurred in large-size fish than small size. Similarly higher parasitic infestation occurred in adults as compared to juveniles. This finding was compared with the reports of (Tasawar et al., 2009), who studied that underweight fish were heavily parasitized by L. cyprinacea as compared to obese weight. However, parasite infestation was increased with an increase in fish length as compared to small size fish due to their more surface area for attachment. This work is also homologous with the finding of (Raissy et al., 2013), who report that L. cyprinacea with regards to length and weight was not statistically significant at (P > 0.05). This research was done by (Roca-Geronès et al., 2020), who concluded that A. simplex is inversely proportioned to the size of a fish host. This study was homologous with the research of (Eissa et al.,, 2018), who stated that Anisakis spp. was more prevalent in obese weight (age) fish than underweight (Allumma et al., 2011). concluded that, among the organs examined only the skin and gills were found to be infected but, there were no parasites found in the intestine.

In this study, two parasites namely Anisakis simplex (Nematoda) and Lernaea cyprinacea (Maxillopoda) species were identified as belonging to different taxonomic groups from 100 fish samples. A total of 03 parasites were identified from fish samples. In this regard, L. cyprinacea (3.5%) was the most commonly found parasite followed by A. simplex (2.5%). The higher prevalence noted of L. cyprinacea was recognized since this parasite was mostly found in the summer season. This study coincides with the finding of (Tasawar et al., 2009), who reported four species of Lernaeids ectoparasites from fish seed hatchery at Mian Channu, which found that L. cyprinacea was the most common (26.67%) parasite among the other Lernaeids ectoparasites. Similarly, (Perveen and Ullah, 2013) during the study on ectoparasites among indigenous and exotic freshwater carp, reported that L. cyprinacea was the most prevalent ectoparasites. This research agrees with the study of (Tasawar et al.,, 2009), who found that the overall prevalence of lernaeids parasites of Catla catla has been reported for L. cyprinacea was (26.67%), L. polymorpha (25.83%), L. oryzophila (4.17) and L. lophiara (2.50%), respectively. Another research was reported by (Tasawar and Arshad, 2001), who concluded that the prevalence of various parasites species on silver carp was L. cyprinacea (9.16%), L. polymorpha (6.66%), L. ctenopharyngodonis (2.5%), L. arcuata (1.6%), L. oryzophila and L. lophiara (2.5%) and (3.33%) respectively. This study compared with the reports of (Wu, 2019), who stated that the prevalence rate of Anisakis third-stage larvae was found of 75% in bummalo, 33.3% in ribbon fish, 25% in saury, and 10% in small yellow croaker fish respectively. This research is homologous with the finding of (Gaglio et al., 2018), who concluded that the Anisakis spp. has prevalence values were 5.4% in Vinciguerria attenuata fish and 2.9% in Electrona risso fish samples.

The current research also showed the histopathological examination of the skin, gills, intestine, and liver of infected and non-infected fish. Histopathological findings were examined in N. tilapia-infected fish by A. simplex, including sloughed-off gill’s epithelium and necrosis in gills lamella, congestion in blood vessels, and hydrophic degeneration of the liver (Fig. 5), Leucocytes infiltration in the intestine (Fig. 6), desquamation of epidermis and necrosis of dermis of skin were observed (Fig. 7). This finding contradicts the reports of (Asgharnia et al., 2022), who concluded that degenerative changes and atrophy occurred in the liver of fish due to Anisakis larvae. Another study was conducted by (Buchmann and Mehrdana, 2016), who reported that the North Atlantic cod liver was heavily infected with third-stage larvae of A. simplex. Infected G. carp fish by L. cyprinacea, showed histopathological findings, including congestion in blood vessels of gills; hydropic degeneration and pyknotic nucleus of the liver, intestine showing normal, desquamation of epidermis and dermis of skin were examined histo-pathologically. This research was compared with the study of (Tufail et al., 2017), who reported that L. cyprinacea-infected fish skin sowed ulceration. The skin bursts up to lower connective tissues from the epidermis and cutis with the infiltration of leukocytes involving lymphocytes, neutrophils, and eosinophils (Furtado et al., 2019). Contradictory to the previous research, most of the internal organs of small fish were invaded by L. cyprinacea. It showed that L. cyprinacea is a more dangerous parasite, especially in juvenile C. catla fish. (Bilal et al., 2021) and (Jerônimo et al.,, 2022) corresponded to our results that the L. cyprinacea-infected fish was traumatic. The parasite penetration attachment site was red and necrotic.

From our results, it was concluded that the present research revealed the occurrence of A. simplex and L. cyprinacea. In the case of L. cyprinacea, relatively high-water temperature provides a suitable condition for reproduction. L. cyprinacea parasites showed slightly a higher prevalence rate as compared to A. simplex. It was found that both parasites occurred at any stage of fish life. They occurred in both male and female fishes. This study determined that 02 parasitic species occurred in fish, which were not earlier observed parasitological in this region. Thus, the findings of this research are estimated to contribute to future studies to stop illegal hunting as overhunting puts fish under stress, thus prone them to parasitic attack. They give Special education to fish biologists for the identification, prevention, and treatment of the infected fish as well as the water body.

Conflict of Interests:

The authors declared that there is no conflict of interest.

Author’s contribution

R. Akhtar and N.U. khan has designed the study. R. Akhtar, N.U. khan and I.A. Khan conducted the experiments. R. Akhtar, N.U. khan and W. Ahmed drafted the manuscript and then S. Sharif, M.H. Ahmad, and I.A. Khan analyzed the data. All authors critically reviewed the manuscript for intellectual content and gave final approval for the final version.

ACKNOWLEDGMENTS

We are thankful to the Higher Education Commission, who supports this research and I also extend thanks to the Department of Pathology, University of Veterinary and Animals Sciences Lahore, Pakistan. This is approved by the ethical Committee and not duplicated anywere.

Data Availability Statement:

The data is available from the corresponding author

Adroher-Auroux, Benítez-Rodríguez (2020) Anisakiasis and Anisakis. An underdiagnosed emerging disease and its main etiological agents 132:535 – 45
Akinsanya H, Adeogun (2008) Gastrointestinal helminth parasites of the fish Synodontis clarias (Siluriformes: Mochokidae) from Lekki lagoon, Lagos, Nigeria. Revista de biología tropical 56(4):2021-6
Allumma, Idowu (2011) Prevalence of gills helminth of Clarias gariepinus in Baga side of Lake Chad. J Appl Sci Environ Manage 15(1)
Allumma I, Management (2011) Prevalence of gills helminth of Clarias gariepinus in Baga side of. Lake Chad 15(1)
Asgharnia M, Roohi et al (2022) ,., First Record of Anisakis sp. Third-Stage Larvae (Nematoda) Occurrence, as a Human Risk, in Pacific Jack Mackerel from a Fish Processing and Packaging Plant in the North of Iran 11(2):95 – 9.
Bao P, Strachan et al (2019) Human health, legislative and socioeconomic issues caused by the fish-borne zoonotic parasite Anisakis. Challenges risk Assess 86:298–310
Bilal A, Atique et al (2021) Lernaeid parasites prevalence in commercial freshwater fish species at various fish. farms Pakistan 84:e253868
Buchmann, Mehrdana (2016) Effects of anisakid nematodes Anisakis simplex (sl), Pseudoterranova decipiens (sl) and Contracaecum osculatum (sl) on fish and consumer health. Food Waterborne Parasitol 4:13–22
Chakona R, Kadye (2019) First record of Lernaea cyprinacea (Copepoda: Lernaeidae) on an imperilled endemic anabantid, Sandelia bainsii (Teleostei: Anabantidae), from the Eastern Cape province. South Afr 44(2):183–187
Chen, Shih (2015) Occurrence and prevalence of fish-borne Anisakis larvae in the spotted mackerel Scomber australasicus from Taiwanese waters. Acta Trop 145:61–67
Debenedetti M, Trelis et al (2019) Prevalence and risk of anisakid larvae in fresh fish frequently consumed in Spain: An Overview. Fishes 4(1):13
Eissa S, Ismail et al (2018) Identification and prevalence of Anisakis pegreffii and A. pegreffii× A. Simplex (ss) hybrid genotype larvae in Atlantic horse Mackerel (Trachurus trachurus) from some North African Mediterranean coasts. Egypt J Aquat Res 44(1):21–27
Furtado C, Figueredo et al (2019) Histological and hematological alterations of silver catfish Rhamdia quelen highly parasitized by. Lernaea cyprinacea 135(2):157–168
Gaglio B, Costa et al (2018) Anisakis spp. larvae in three mesopelagic and bathypelagic fish species of the central Mediterranean Sea. Parasitol Int 67(1):23–28
Golden Araújo, Caldeira et al (2023) ,., Raw fish consumption in Portugal: Commonly consumed fish species and associated risk factors for anisakiosis 145:109457.
Hua Z, Zou et al (2019) Morphology is not a reliable taxonomic tool for the genus Lernaea: molecular data and experimental infection reveal that L. cyprinacea and L. cruciata are conspecific 12:1–13
Innal, Avenant-Oldewage (2012) Occurrence of Lernaea cyprinacea on mosquito fish (Gambusia affinis) from Kundu Estuary (Antalya-Turkey). Bull Eur Assoc Fish Pathol 32(4):140–147
Jerônimo D, Cruz (eds) (2022) Bertaglia, Fish parasites can reflect environmental quality in fish farms 14(3):1558-71
Justus O, Bebe (2012) Determinants of Adoption of Management Interventions in Indigenous Chicken Production in Kenya. Afr J Agricultural Resource Econ (AFJARE) 7:39–50
Koyun U, Mart (2015) First Record of Lernaea cyprinacea L. 1758 (Copepoda: Cyclopoida) on Cyprinion macrostomus Heckel, 1843 from Eastern Anatolia, Turkey. Biharean Biol 9:44–46
Mathews (2004) Design of Experiments with MINITAB: Quality press
Nur B, Fadli et al (2022) Lernaea cyprinacea Linnaeus, 1758 (Copepoda: Lernaeidae) infection on Betta rubra Perugia, 1893 (Anabantiformes: Osphronemidae) from Aceh Province. Indonesia 31:e020421
Oliveros (2007) VENNY. An interactive tool for comparing lists with Venn Diagrams
Perveen, Ullah (2013) Ectoparasites of indigenous and exotic fresh water carp fish (Cypriniformes: Cyprinidae) from Charbanda and Tarbela, Khyber Pakhtunkhwa, Pakistan. Pak Am J Res Commun 1(9):255–269
Rahmati-Holasoo et al (2023) Marandi, Shokrpoor,., Clinico-histopathological and phylogenetic analysis of protozoan epibiont Epistylis wuhanensis associated with crustacean parasite Lernaea cyprinacea from ornamental fish in Iran 13(1):14065.
Raissy S, Rashedi et al (2013) Investigation of a parasitic outbreak of Lernaea cyprinacea Linnaeus (Crustacea: Copepoda) in Cyprinid fish from Choghakhor lagoon. Iran J Fisheries Sci 12(3):680–688
Raja P, Avunje et al (2023) Natural infestation of an anchor worm, Lernaea sp. in cage culture of Asian Seabass, Lates calcarifer juveniles and its control using an anti-parasitic drug. emamectin benzoate 47(2):306–318
Roca-Geronès S, Godínez-González et al (2022) Anisakis and Hysterothylacium species in Mediterranean and North-East Atlantic fishes commonly consumed in Spain. Epidemiol Mol morphometric discriminant Anal 325:108642
Slaoui, Fiette (2011) Histopathology procedures: from tissue sampling to histopathological evaluation. Springer, Drug safety evaluation, pp 69–82
Tasawar, Arshad (2001) Seasonal prevalence of Copepod Ectoparasites (Lernaea sp.) of silver carp (Hypophtylmicthys molitrix). Punj Uni J Zool 15:49–54
Tasawar N, Lashari et al (2009) Incidence of lenaeid parasite in Catla catla on a fish farm. Sarhad J Agric 25(2):285–289
Tasawar Z, Lashari et al (2009) The prevalence of lernaeid ectoparasites in grass carp. (Ctenopharyngodon idella) 29(2)
Thrusfield (2007) What sample size should be selected. Veterinary Epidemiology 232(8)
Trilles R, Rameshkumar (2011) A checklist of the Cymothoidae (Crustacea, Isopoda) recorded from Indian fishes. Acta Parasitol 56(4):446–459
Tufail Q, Khan et al (2017) Prevalence and effects of Lernaea cyprinacea (anchor worm) on the growth, skin histopathology and hematology of Catla catla. Iran J Fisheries Sci 16(1):341–351
Wu (2019) Identification and occurrence of Anisakis larvae from Marine Fish in Changsha City. Hunan province. China 852–859

Download PDF

Reviewers agreed at journal
04 Aug, 2024
Reviewers invited by journal
30 Jul, 2024
Editor assigned by journal
29 Jul, 2024
First submitted to journal
28 Jul, 2024

You are reading this latest preprint version

Spectrum of Zoonotic Fish Parasites in Lahore, Pakistan: An Anatomopathological analysis for tissue damage

Status:

Version 1

Abstract

Figures

Statement of Novelty

INTRODUCTION

MATERIALS AND METHODS

Experimental design:

Dissection procedure:

Parasites identification:

Prevalence rate determination:

---------------------------------- × 100

Histopathological examination:

Sample size calculation:

Statistical analysis:

RESULTS

DISCUSSION

CONCLUSION

Declarations

Conflict of Interests:

ACKNOWLEDGMENTS

Data Availability Statement:

References

Status:

Version 1