Treatment of Lernaeasis in Carps With Thunder: An Organophosphate

See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/259354178 Treatment of Lernaeasis in Carps With Thunder: An Organophosphate Article in Pakistan journal of zoology · March 2012 CITATIONS READS 3 74 3 authors, including: Zafar Iqbal Khalid Pervaiz 53 PUBLICATIONS 77 CITATIONS 24 PUBLICATIONS 27 CITATIONS University of the Punjab, Lahore,Pakistan SEE PROFILE Deapartment of Fisheries Government of the P… SEE PROFILE All content following this page was uploaded by Khalid Pervaiz on 28 October 2014. The user has requested enhancement of the downloaded file. All in-text references underlined in blue are added to the original document and are linked to publications on ResearchGate, letting you access and read them immediately. Pakistan J. Zool., vol. 44 (2), pp. 415-420, 2012. Treatment of Lernaeasis in Carps With Thunder: An Organophosphate Zafar Iqbal*, Iffat Rani and Khalid Pervaiz Department of Zoology, University of the Punjab, New Campus, Lahore. P. code 54590, Pakistan Abstract.- The carps; Labeo rohita Hamilton, 1822, Catla catla Hamilton, 1822, Ctenopharyngodon idella Valenciennes, 1844, Hypophthalmichthys molitrix Richardson, 1845 and Aristichthys nobilis Richardson, 1845 were collected from earthen ponds reared under semi-intensive conditions. The parasitic examination of these fishes showed that all fish species were infected with crustacean copepod parasite Lernaea cyprinacea and other Lernaea spp. C. catla, L. rohita and C. idella had higher prevalence (62.5%, 68.75% and 76%, respectively) and mean intensity (12.1, 8.45 and 8.84 parasite/fish, respectively) compared to H. molitrix (9.1% and 3.0 parasite/fish). A locally available drug “Thunder” an organophosphate (2, 2 dichlorovinyl dimethyl phosphate) was used to treat lernaeasis in these fish species. Three concentrations (0.10, 0.15 and 0.20ppm) of the Thunder were sprayed on the surface of the infected fish ponds to eradicate lernaeasis. The 0.20ppm dose proved to be the most effective in controlling the infection, without harming the fish. The water quality parameters remained in normal range and suitable for the fish after treatment indicating that ‘Thunder’ has no harmful effects on water quality in fish pond. Keywords: carps, Lernaeasis, treatment, Thunder / DDVP. INTRODUCTION The carp fish farming in private sector in some parts of Pakistan especially in Punjab has increased in recent years. In 2010, the Directorate General of Fisheries Punjab (DGOFP) reported establishment of more than 7829 fish farms covering approximately 45650 acres in Punjab. The growth of fish culture has also highlighted many issues of fish health, such as fish diseases and their management. Horvath et al. (1992) discussed various factors for fish disease occurrence in ponds, such as stocking density, aquaculture technology application and pollution discharge. In addition, numerous pathogens, nutritional deficiency, sudden changes in biotic and abiotic factors are causes of diseases in pond fishes (Roberts, 1989). Disease causes mortality, low growth, poor production and loss of fecundity (Bauer et al., 1973). Hoole et al. (2001) gave an account on carp diseases. According to Iqbal et al. (2000, 2001) abdominal dropsy, lernaeasis, anoxia and fungal infection are the most commonly occurring diseases in pond fishes in Punjab. ______________________________ * Corresponding author: dr.zafariqbal_pu@yahoo.com 0030-9923/2012/0002-0415 $ 8.00/0 Copyright 2012 Zoological Society of Pakistan. Adult female anchor worm, Lernaea cyprinacea causes lernaeasis. It lacks host specificity to such an extent that it can infect all freshwater fishes (Schmidth, 1990). The red sores caused by L. cyprinacea on fish skin are prone to secondary bacterial and fungal infection (Putz and Bowen, 1964 and Iqbal unpublished data). The infected fish become listless and weak (Iqbal et al., 2001). In Africa rapid increase in Lernaea burden has been associated with growing environmental stress. Oldewage (1993) reported high prevalence of Lernaea spp. in tilapia of Lake Victoria. Heavy infection with Lernaea spp. has resulted mortality in golden perch, Macquaria ambigua Richardson, 1845 murray cod, Maccullochella peelii Mitchell, 1838 and silver perch, Bidyanus bidyanus Mitchell, 1838, broodstock held in pond (Callinan, 1988). Low level infection in river fishes and mortality due to lernaeasis in farmed fish has been reported by Gabrielli and Orsi (2000). A fairly comprehensive review of control measures against Lernaea spp has been published by Hoffman and Mayer (1974). Most control measures involve application of chemicals, sometime on a large scale in ponds. Chemicals such as Formalin (Putz and Brown, 1964), Dylox (Hoffman and Mayer, 1974), Dimilin (Burtle and Morrison, 1986) and Dipterex (Minhas et al., 2001) are commonly used for the treatment of lernaeasis in 416 Z. IQBAL ET AL. commercial fisheries. Among the existing veterinary drugs and chemicals, the organophasphate (OP) insecticide and acaricide dichlorvos have been extensively used to treat sea lice infestations by Lepeophtheirus salmonis and Caligus elongates in the Atlantic salmon Salmo salar culture. Organophasphate is relatively non-persistent and undergoes fast and complete hydrolysis and metabolizes in most environments (WHO, 1989). Dipterex is very expensive chemical and its availability is not certain especially in remote rural areas in Punjab. The present study was aimed to test a comparatively cheap and locally available drug, “Thunder” an organophosphate, (2, 2 dichlorovinyl dimethyl phosphate or DDVP), to treat Lernaeasis in five species of carp L. rohita, C. catla, C. idella, H. molitrix and A. nobilis in fish ponds. MATERIALS AND METHODS The diseased fishes were collected from four ponds, at the University of the Punjab Research Fish farms during the spring of 2005. Each fish was wrapped in plastic bag laid in crushed ice and brought to laboratory. The fishes were weighed and measured. All fishes were examined thoroughly for the presence of any parasites or lesions visible to naked eye and also with the help of magnifying glass. The parasites were examined under microscope at x10 magnification. Lernaea spp were collected and preserved in specimen bottles in 70% alcohol for subsequent examination. “Thunder” is extensively used insecticide in Pakistan. Its active ingredient is dichlorvos, which contains DDVP (2, 2 dichlorovinyl dimethyl phosphate) and other ingredients (58.33% w/w). Thunder is supplied in 500 ml packing in plastic bottles by Welgreen Chemicals (Pvt.) Ltd. Lahore. Three fish ponds (1, 2, 3; stocking density of 350 fishes in each pond) having infected fishes (infection level in these ponds before treatment is given in Table I) were selected for treatment of lernaeasis. The area of the ponds and volume of pond water was calculated according to Minhas et al. (2001). The dimension of infected ponds were 175 x 85 x 4.5 feet (ponds 1-3) and 175 x 100 x 4.5 feet (pond 4). The desired quantity of “Thunder” was dissolved in 15-litre clean water in hand sprayer and sprayed on the surface of each pond (0.10ppm in pond 1; 0.15ppm in pond 2 and 0.20ppm in pond 3, respectively, according to Burtle and Morrison (1987). The pond 4 was used as control and no treatment was applied. Treatment in these ponds was repeated in seven days interval and continued for three weeks. One dose was given at the start of each week. Dose 1 on first week, dose II on second week, and dose III on the last week respectively, was repeated after every seven days interval and continued for 21 days. The fishes were examined after each treatment. Pond water temperature, dissolved oxygen and pH were measured on site with DO meter YSI (Model 57) and digital pH meter on sampling day before and after the treatment in these ponds. Water samples were taken in sterilized one liter glass bottles for subsequent analysis at water quality management laboratory, Fisheries Research and Training Institute, Lahore. RESULTS Infection of fish with Lernaea cyprinacea In total 71 fish were examined to assess the level of infection in the stock. These fishes were heavily infected with Lernaea cyprinacea and other Lernaea spp. The parasites were attached to skin, fins and abdomen of the infected fish. The attachment of parasite on fish caused lesions on the skin. The infected fish showed discoloration of skin, darkening of the scales and poor growth and health. The detail of the fish examined and infected is given in Table I. Table I indicates that all four ponds were infected and had high infection of Lernaea, viz., pond 1, 62.5%; pond 2, 42.1%; pond 3, 76.5% and pond 4, 63.2 %. Table I also indicates 69% prevalence in L. rohita; 62.5% in C. catla; 76% in C. idella; 9% in H. molitrix and 67% in A. nobilis. The mean intensity was 8.45 in L.rohita; 12.1 in C.catla and 8.84 in C.idella. The number of parasites per infected fish varied from 4 to 20 in L.rohita; 3 to 32 in C.catla and 1 to 26 in C.idella in these ponds. Overall infection was about 61% and mean intensity 9.0. Treatment of lernaeasis The treatment of lernaeasis in these infected LERNAEASIS TREATMENT WITH THUNDER Table I.- Prevalence and mean intensity of Lernaea spp. in different ponds. Fish Weight (g) Pond 1 No. of fish Exa. Infec. Pond 2 No. of fish Exa. Infec. Pond 3 No. of fish Exa. Infec. Pond 4 No. of fish Exa. Infec. 417 Total Exa. Infec. Prevalence % Mean intensity L. rohita 2431015 4 2 5 3 3 3 4 3 16 11 69 8.45 C. catla 135235 2 1 2 1 7 5 5 3 16 10 625 12.1 C. idella 237786 6 6 8 3 5 4 6 6 25 19 76.0 8.84 H. molitrix 218324 4 1 3 - 1 - 3 - 11 1 9 3 A. nobilis 422781 - - 1 1 1 1 1 - 3 2 67 1 Prevalence Mean intensity 62.5% 42.1% 76.2% 63.2% 60.56% 8.5 8.12 9.40 9.5 9.0 Exa, examined; Infec, infected. ponds was carried by spraying “Thunder” on the surface of the ponds. The first dose of “Thunder” was applied to pond 1, 2, and 3 at a concentration of 0.10ppm, 0.15ppm and 0.20ppm, respectively. The treatment continued for 3 weeks. Table II indicates that fishes were still infected after dose-1 and dose11 of “Thunder” application. The infection was 85.7% in pond 1; 52.6% in pond 2; 50% in pond 3 after dose-1. All fish species showed infection even after application of dose-11 which was 64.2% in pond 1; 32.5% in pond 2 and 18.7% in pond 3. There was no infection after the application of dose-III in pond 2 and pond 3. However, 5.5% infection was recorded in Pond 1. The average number of parasites ranged from 8-19 (pond 1); 6-8 parasites (Pond 2) and 6 parasites per infected fish (Pond 3) after dose I. Table III, shows that infection was present in ponds 1, 2 and 3 even after application of dose II. However, average number of parasites reduced to 6, 5 and 4 in these ponds, respectively. In case of Pond 3, infection reduced after dose II application, only three fish specimens (two C. idella and one C. catla) were found with less infection. The fishes were examined after doseIII application and all the fishes showed no infection. Whereas in control pond, fish infection persisted even mortality of one fish specimen occurred due to lernaeasis. Water quality parameters The water analysis of the ponds before and after treatment (Table III) shows that “Thunder” does not have adverse effect on water quality of the ponds after treatment. DISCUSSION Treatment of L. cyprinacea involving baths and dips in a container is a prolonged and timeconsuming process. Direct application of chemicals in a pond is a convenient and effective method and also avoids fish from handling stress. This study showed that “Thunder” in various concentrations is an effective drug to treat and eradicate lernaeasis in five species of carp. However, 0.20ppm is considered as the most effective concentration and has no harmful effect on fish. The infective stages of Lernaea in the ponds could not survive after repeated application of “Thunder”. Also, it minimized fresh infection and re-infection in the fishes in each pond. The fishes observed after treatment appeared to be healthy. Abidi et al. (2000) reported that Thunder at 0.5ppm is not harmful to fish life, flesh quality or aquatic environment. At 0.5 and 1.5ppm Thunder has no adverse effect on growth of micro-algae in freshwater (Ayub et al., 2000). 418 Table II.- Z. IQBAL ET AL. Prevalence and mean intensity of Lernaea spp. after treatment with “Thunder” in three fish ponds. Pond Fish 1 L. rohita C. catla C. idella H. molitrix A. nobilis Total Prevalence (%) Mean intensity 4 1 6 2 1 14 4 1 5 1 1 12 85.7 7.0 3 2 6 3 14 2 2 5 0 9 64.2 3.5 4 6 5 2 1 18 1 1 5.5 2 2 L. rohita C. catla C. idella H. molitrix A. nobilis Total Prevalence (%) Mean intensity 5 3 9 2 19 4 3 7 1 15 52.6 5.0 4 4 6 2 16 0 2 4 0 6 32.5 3 2 4 6 3 15 0 0 0 3 L. rohita C. catla C. idella H. molitrix A. nobilis Total Prevalence (%) Mean intensity 2 7 4 3 16 1 5 2 0 8 50 3.5 5 1 6 4 16 0 1 2 0 3 18.7 2.0 6 3 5 3 1 18 0 0 0 Dose I (0.10 ppm) Examined Infected Kashara (1962) used organophosphate trichlorophan for effective eradication of anchor worm larva at 0.20ppm in two days and in one day at 0.5ppm. It destroyed free floating larvae and adult parasites. Putz and Bowen (1964) used formalin at 250ppm to treat larval stages of L. cyprinacea. However, this concentration is considered unsafe for fish health. Lernaea spp. was killed by weekly application of 0.27ppm of Lexone (Gopalkrishan, 1964). Hoffman and Mayer (1974) listed Benzene Hexachloride as effective against Lernaea parasites on Carassius auratus. Routine application of Masoten by spraying on ponds (0.2-0.5ppm) twice monthly during summer has also been recommended but more as a precautionary measure in Japan (Kabata, 1985). In Indonesia, Dipterex was used successfully at 0.5ppm for three days to treat lenaeasis (Kabata 1985). Chinese aquaculturists eradicated Lernaea after Trichlorophon was used at rate of 2.0ppm for 24 hours (Kabata, 1985). Burtle and Morison (1987) concluded that Dimilin applied Dose II (0.15 ppm) Examined Infected Dose III (0.20 ppm) Examined Infected at 10ppb in ponds was effective to kill Lernaea and other copepods in golden shiner, Notemigonus crysoleucas Mitchell, 1814. Minhas et al. (2001) applied two doses of Dipterex at 0.2ppm in 2 weeks intervals. This dose was found effective to eliminate L. cyprinacea in culturable carps. The application of ‘Thunder’ at 0.20ppm in the present study is comparable to Minhas et al. (2001) and is evident from low infection in pond 3 after dose-II and no infection after dose-III. The effect of “Thunder” at 0.20ppm to H. molitrix seems to be very quick as compared to other fish species as observed in this study (Table II). Hemaprasanth et al. (2008) suggested a single intramuscular administration of the drug Doramectin at 200 ppb/kg body weight for treatment of heavily infected adults and brood stock of carps with L. cyprinacea. Although Lernaea induced fish kill are not common (Shariff and Roberts, 1989), however, presence of Lernaea has been observed in case of LERNAEASIS TREATMENT WITH THUNDER 419 Table III.- Water analysis of fish pond water before and after treatment. Pond Temp. (°C) P-1 * ** P-2* ** P-3* ** P4* *** 24 30.5 24 30 23 30 23 30.5 pH DO (mg/ L) Bicarbonate as CaCO3 (mg/L) Total alkalinity as CaCO3 (mg/L) Calcium as CaCO3 (mg/L) Total hardness as CaCO3 (mg/L) Chloride (mg/L) TDS (mg/L) 7.6 7.8 7.4 7.5 7.6 7.5 7.5 7.7 5.6 4.9 5.5 5.1 5.6 5.0 5.5 4.9 483 457 470 486 474 441 337 436 483 491 470 520 474 499 337 470 108 115 100 111 104 108 92 123 211 204 200 204 207 200 180 230 55 79 68 94 78 69 72 64 715 618 650 618 715 618 650 585 *Before treatment, **After treatment, *** Analysis after three weeks in P-4 (control pond). culturable carps in Punjab (Iqbal et al., 2001) and in the present study. To maximize fish production in an intensive and semi-intensive culture system the relationship between fish species cultured and crustacean copepod causing infection and diseases must be understood (Piasecki et al., 2004). “Thunder” is ecologically safe due to its biodegradability. It remains in water for 36h before it disintegrates and completely disappears from water (Abidi et al., 2000). “Thunder” at a dose of 0.5 and 1.5ppm has no adverse effects on growth of an important micro-algae Scenedesmus quardicauda, natural food of filter feeding fish (Ayub et al., 2000). All water quality parameters in our study were suitable for fish culture and within the normal range for warm water fish culture as defined by Boyd and Tucker (1998). The treatment of lernaeasis with “Thunder’ at 0.20 ppm (dose-III) has shown the promising results in three weeks. There was no indication that fish were likely to be adversely affected by the use of “Thunder” at this level. Further trial with higher doses may probably reduce the treatment time, however, this remains to be tested. Since, ‘Thunder” has been reported to be safe for aquaculture industry at a dose of 0.5ppm (Abidi et al., 2000) we suggest that precautionary measures be taken at this level of application during fish rearing to minimize infection and further, this treatment level may increase stress to fish in pond. In our study, crustacean copepod parasites L. cyprinacea and other Lernaea spp. are more sensitive to “Thunder” than fish and differences in sensitivity impact the characterization of risk. The actual extent of mortality would depend on the rate at which “Thunder” is applied, its concentration, the degree of mixing that occurs in the water and the rate of breakdown and dissipation. These processes suggest that adverse effects in sensitive species are plausible. No effects are likely in less sensitive species. 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