The Journal of Basic & Applied Zoology (2016) 74, 62–67 H O S T E D BY The Egyptian German Society for Zoology The Journal of Basic & Applied Zoology www.egsz.org www.sciencedirect.com Some ecological and toxicological studies on crayfish, Procambarus clarkii at Sharkia Governorate, Egypt Samah M Abdel-Kader Plant Protection Research Institute, Agric Res Center, Dokki, Giza, Egypt Received 17 June 2016; revised 13 August 2016; accepted 20 August 2016 KEYWORDS Procambarus clarkii; Burrowing behavior; Furadan Abstract The Louisiana red swamp crayfish, Procambarus clarkii, which was introduced into Egypt in the1980s, has since become widespread across the country Burrowing behavior of P clarkii, was investigated in irrigation canal of three different sites (Abou-Kabir, Abou-hammad and Belbies) at Sharkia Governorate from January 2014 to December 2015 Most burrows were simple in their morphology and few of them had two types of chimney (plugged and open) Open chimney was present at 100 cm of the edge side of irrigation canal, while plugged cap chimney was located inside the agriculture soil Result indicated that a great number of P clarkii were observed migrating across roadways to find the main irrigation canal, during their crossing many of them were killed by cars The highest mean numbers of burrows and population density of P clarkii were recorded at Abou-Kabir followed by Abou-hammad while Belbies was the lowest one in this respect during two successive years of 2014 and 2015 Furadan 10G is routinely applied in Egypt rice fields was examined in the laboratory against P clarkii in trial to control its explosive populations in certain closed habitats Data showed that after 96 h, at the higher concentrations (70 ppm) 100 and 90% of mortality were recorded for adult males and females of P clarkii, respectively while at lower concentrations (50 ppm) the mortality was 70 and 60% for both sexes respectively LC50 of furadan was 58.26 and 63.60 ppm for adult males and females, respectively Ó 2016 The Egyptian German Society for Zoology Production and hosting by Elsevier B.V This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/) Introduction The red swamp crayfish, P clarkii (Girard, 1852) native to the south central United States (Louisiana) and north-eastern Mexico, has been transplanted world-wide (Huner, 1988; Gherardi and Holdich, 1999) E-mail address: samah_kader@yahoo.com Peer review under responsibility of The Egyptian German Society for Zoology The introduction of the invasive P clarkii in Egypt in early 1980s for aquaculture (Ibrahim et al., 1995) caused, and is still causing changes in both the structure of ecological communities and the function of ecosystems It has been successfully established in various sites of Nile River and its branches, from northern Delta to Assuit (Saad and Emam, 1998) P clarkii has active dispersal capabilities which enable it to successfully expand its range to new areas, when other environmental conditions are favorable (Correia, 2002), such as a suitable soil type, location and permanence of the water table and food http://dx.doi.org/10.1016/j.jobaz.2016.08.001 2090-9896 Ó 2016 The Egyptian German Society for Zoology Production and hosting by Elsevier B.V This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/) Ecological and toxicological studies on crayfish supply (Hogger, 1988), enough cover vegetation (John and Avault, 1981), and low predators (Correia, 1993), may play an important role in its distributions This crayfish invades most of the wetland areas and rice fields, causing serious damages to drainage systems and rice crop as consequence of its digging activities (Burton et al., 1980; Brown, 1994; Soliman et al., 1998) which result in water leakage, and damage fields (Yue et al., 2010a) Moreover, it causes a lot of damage to fisheries of the Nile River possibly by attacking the fry and young fish and damaging the nets of fishermen (Ibrahim et al., 1995) Additionally, this damage may lead farmers to use pesticides to control P clarkii (Hobbs et al., 1989; Anasta´cio and Marques, 1995; Jime´nez et al., 2003) However, once introduced to a favorable habitat, it is hardly eliminated, (Hobbs et al., 1989; Anasta´cio et al., 2000) Control with chemical substances is still a useful method today, particularly over certain limited aquatic areas against the crayfish expanding populations of these substances is furadan 10G Furadan 10G is routinely applied to Egypt rice fields at the concentration of 4.0 kg/acre (1.62 kg/ha) to control insect pests Moreover, furadan has been found to be normally degraded rapidly (Godan, 1983) However, little information is available about the toxicity of furadan on aquatic macro-invertebrates (Alves et al., 2002) The objectives of these research were (1) studying the expansion and population density of P clarkii in Sharkia Governorate and discussing some aspects dealt with its spatial distribution (2) to investigate, the toxicity of carbamate pesticide furadanÒ 10G on adults crayfish, P clarkii (males and females) in the laboratory and to determine the lowest lethal concentration of furadan producing death for a population of P clarkii Study areas and methods Description of the study areas The field experiments were carried out in irrigation canal of Abou-Kabir (Gynabeit Bahr Fakous), Abou-hammad (kafer Abou-ayad) and Belbies (El-Damshyshia) districts at Sharkia Governorate from January 2014 to December 2015 AbouKabir canal is approximately about km in length, its width m with depth varied between and m The bottom is sandy clayey loam with extensive field crops and had an abundant bank cover vegetation due to the permanence of the water table (Gynabeit Bahr Fakous) such as Salix tetrasperma, Pulchea dioscroidis, Desmotachya bipinnata, Ipomoea carnea as well as Bermuda grass, Cynodon dactylon L and Vossia cuspidata The length of Abou-hammad irrigation canal is about km, its width ranges from to ms with depth varying between and m The bottom is characterized by sandy clayey silt in sediment nature which is surrounded by green crop fields with a little grass communities (Ludwigia stolonifera, Cyperus alopecuroides, Ipomoea carnea and Mentha microphylla) around the edges of the canal While, the Belbies irrigation canal has very low water levels or is completely dry during a large part of the year 2015 and it is approximately about km in length, the water depth average m and its width from to m This area is clay in nature and usually cultivated with the different types of field crops and vegetables, but had no cover vegetations around its edge The range of water quality conditions where P clarkii is found are: pH 63 ranged between 7.3 and 7.8, dissolved oxygen concentration 400–1400 ppm, salinity between 100 and 300 ppm, and water temperature 14–26 °C Collection of specimens The number of burrows (per m2) and population density (Gobbia/m2) were monthly determined in three selected sites at Sharkia Governorate between January 2014 and December 2015 Specimens were captured using small-mesh (15 mm in diameter), cylindrical traps (Gobbia) set across stream channel The traps were set in the afternoon and retrieved 24 h later All catches recorded were used to estimate the population density The collected specimens were transferred to the laboratory for further investigation Laboratory procedure Tested pesticide Furadan 10G is a granular carbamate insecticide with a 10% concentration of the active ingredient carbofuran (CF, 2,3-di hydro-2,2-dimethyl-7-benzo-furanil methyl carbamate) which was obtained from Al-Gomhouria Chemical Co Tested animals and acclimation procedures Adults of red crayfish P clarkii (22–29 gm average weight and 8.5–9 cm total length) were collected from the same study areas and carried immediately to the laboratory in well-aerated large plastic tanks (30 Â 50 Â 30 cm) and acclimated for one week Acclimation tanks maintained at a temperature of 21–23, a photo period of 12 h light and housed in 10 cm deep aged tap water with a daily diet of cucumber Water was changed once every day Feeding was suspended 48 prior to the experiment Acute toxicity determination Furadan 10G is routinely applied to Egypt rice fields at the concentration of 4.0 kg/acre to control insect pests Our tests were conducted to determine crayfish mortality starting with the recommended concentration applied in rice fields followed by a geometric series of different pesticide concentrations to determine the median lethal concentration (LC50) for both adult males and females of P clarkii To determine the median lethal concentration (LC50) of furadan, stock solution was prepared using dechlorinated tap water as a solvent to achieve desired concentrations Adult crayfish of P clarkii were exposed to three concentrations of furadan (50, 60 and 70 ppm) These concentrations were chosen on the basis of range finding tests The experiment was repeated times, which exposed (10 Â 3) P clarkii to each test concentration, plus control (10 Â 1), for 96 h Crayfish mortality increased with increasing furadan concentration and exposure time The test containers were checked every h over 96 h to record and remove dead animals as soon as observed The criterion for death was the failure of crayfish specimen to respond to prodding with a blunt glass probe The concentration 64 S.M Abdel-Kader causing 50% mortality of the tested animals (LC50) and 95% confidence limits were calculated according to Proban Program (1991) Results and discussion Number of burrows P clarkii constructs deep burrows in every soil type unrelated to time of the year along the shoreline of the irrigation canals (Fig 1) Burrow is simple in its morphology, with one tunnel ranging from 40 to 90 cm and a single entrance, each 1–5 in diameter These findings were quite similar to those previously described by Hogger (1988), Ilhe´u (1996) and Soliman et al (1998) Burrow is marked by two types of burrow cap (open chimney cm in height and plugged cap cm in height) Open chimney present at 100 cm of the edge side of irrigation canal This chimney is thought to minimize water loss from the burrow (Fig 2) A similar observation was recorded for other species, Cambarus fodiens (Cottle) by Williams et al (1974), who found that, the burrows with a chimney are located at a greater distance from the water surface than simple burrows How- Figure Showing a burrow of P clarkii at the lower side of banks of irrigation canals Figure ever, plugged cap chimney is located inside the agriculture soil, where the plugged cap prevents water evaporation from the burrow, which would result in increased p clarkii survival during drought, as previously suggested for both types of open and closed chimneys by Barbaresi et al.,(2004) The number of burrows which is considerably different from one area to another during the two years of the study (2014 and 2015 years) is shown in Table Abou-Kabir district harbored the highest mean number of burrows in winter (67and76 burrows per 50 m2) during 2014 and 2015 , respectively) in comparison with the other two tested areas as 26–34 and 22–12 burrows (per 50 m2) at Abou-hammad and Belbies respectively, while the lowest mean numbers were recorded in summer at the three sites (22–16; 7–10 & 0–0 burrows) during 2014 and 2015, respectively This burrowing activity allows the species to avoid extreme environmental conditions such as low and high temperatures and dehydration also provides refuge from predation (Gherardi et al., 2002; Ilhe´u et al., 2003) or to lay and incubate eggs (Huner and Barr, 1991; Soliman et al., 1998) Population density The seasonal variation in population density of P clarkii in irrigation canals of the three tested districts during 2014 and 2015 are illustrated in Table The obtained data revealed that the abundance of P clarkii is positively correlated with the water permanent (Gynabeit Bahr Fakous), as in Abou-Kabir district which recorded the highest populations of animals (306 & 370 per Gobbia/m2) during 2014 & 2015, where, dense cover of bank vegetation with a low density of predators This finding is in agreement with Grow and Merchant,(1979), Grow (1982) and Hobbs and Whiteman (1991), who assured that the presence of free water has an additional documented effect on the crayfish’s burrowing ability Also, Huner and Barr (1991) reported that a dense beds of grass with a low density of predators has positive effects on the abundance of P clarkii, because erect stems of emergent plants provide food and shelter (Lodge and Hill, 1994; Foster, 1995; Naura and Robinson, 1998) In early spring, we noticed after the dawn, a very large numbers of P clarkii moved leaving their burrows of the field into overland crossing the road way (distance about 12 m) to Showing P clarkii construction of open chimney or burrows cap around the complete burrow Ecological and toxicological studies on crayfish Table Mean number of P clarkii burrows at the three tested areas (per 50 m2) during different seasons of 2014 to 2015 years Locations 2014 Abou-Kabir Abou-Hammad Belbies Grand total Table 65 Total Winter Spring Summer Autumn 67 26 22 36 20 22 40 18 10 165 74 37 276 2015 Total Winter Spring Summer Autumn 76 34 12 44 14 16 10 56 65 192 79 12 283 Mean number of population density of P clarkii at the three tested areas during the period from 2014 to 2015 Locations Seasons Population density/Gobbia Jan Feb Mar Apr Ma Jun Jul Aug Sep Oct Nov Dec Total Average Abou-Kabir 2014 2015 0 0 0 10 43 57 96 104 87 89 35 55 24 33 10 20 306 370 25.4 30.8 Abou-Hammad 2014 2015 0 0 0 15 27 38 82 I00 36 43 26 33 10 22 13 202 262 16.8 21.8 Belbies 2014 2015 0 0 0 0 0 2 10 40 20 90 10 7.5 0.8 Table Showing the effect of different concentrations of furadan insecticide against adult males and females of P clarkii Conc.ppm No of animals Mortality of adults/h Males 50 60 70 Control (0) Total mortality % Mortality Females # $ 24 48 72 96 24 48 72 96 # $ # $ 10 10 10 10 10 10 10 10 3 3 2 0 1 2 10 70 90 100 60 70 90 find the main water course (Gynabeit Bahr Fakous), and to complete the maturation of newly hatched crayfish The maturation needs hormonal induction by the photoperiod, a hydroperiod longer than four months, a temperature above 18 °C, and a pH between and (Gutie´rrez-Yurrita, 1997) This terrestrial behavior was registered by Huner (2002) and Chucholl (2011) who assured that P clarkii can survive the drying up of seasonal water bodies by burrowing or migrating across land to new habitats, in response to poor water quality (McCarthy et al., 2014), or for Seeking reproductive opportunities and dispersal between waterways (Morgan, 1991; Furse et al., 2004) At Abou-hammad, the relatively highest population density was recorded (202 & 262 per Gobbia/m2) during 2014 and 2015, respectively, while, there was a marked reduction in population density through 2014 and 2015 (90 and 10 per Gobbia/m2 respectively) at irrigation canals of Belbies, due to irrigation water leakage (sometimes dried up completely) which resulted in, both young and adult P clarkii failed to dig burrow in dry soil Consequently, they died or became more susceptible to attack by many types of predators, especially Norway rat that infested this site A similar observation was recorded by Emam and Khalil (1995) and Mubarak (2001) The present study shows that the permanent table water, cover vegetation and predation appear to be the most important factor controlling the population density of P clarkii Toxicity test The lethal effects of different concentrations of furadan compound on both sexes of P clarkii at different exposure periods (96 h) are shown in Table Mortality percentages of furadan was dose dependent and the durations of exposure dependent, at the higher concentrations (typical concentration used in Egypt rice fields (4.0 kg/ acre), all of the adult crayfish were killed within one hour The maximum mortality still remained 100 and 90% at concentration 70 ppm for adult males and females respectively, but this activity declined progressively as the concentration decreased (50 ppm) where the mortality was 70 and 60% for both sex respectively No mortality was observed in controls during the experiment LC50 of furadan after 96 h was 58.26 ppm (95% Fuducial Limits (FL): 51.07–63.60) and 63.60 ppm (95% FL: 56.99–65.40) for adult males and females of P clarkii, respectively It was clear that, there was a fairly variation in tolerance between the two sexes of P clarkii This difference may be due to the heterogeneity between tested individual which resulted in difference in their response for toxicity Furadan is known to have a strong toxicity effect against a wide spectrum of agricultural pests through the inhibition of the enzyme cholinesterase (Ware, 1994) 66 The obtained results indicated that the highest toxicity effect was obtained with the typical concentration used in Egypt rice fields (4.0 kg/acre) and at concentration 70 ppm Similar toxic effects were reported by other investigators, Carter and Graves (1973) who found that methyl parathion carbaryl and carbofuran were highly toxic to crayfish, and their 96-h LC50 values were 3.0, 500 and 500 ug/I respectively Ekanem et al (1981) found that, in a water-soil medium, mortality of juvenile Louisiana crayfish was 60% at a concentration of 1.0 kg/acre of active ingredient furadan Rosenthal et al (2005) reported that furadan 5G induced 90% of the adult crayfish mortality that occurred within the first 24 h at 4.0 kg/acre concentrations used in Kenyan rice fields (0.20 kg/acre active ingredient) under laboratory conditions Any comparison between our data with the previous toxicity studies is difficult because there are several commercial formulations of furadan with different concentration of active ingredients of the products or may be due to other methodological differences, or to crayfish size and different test conditions Laboratory studies are not adequate in the control of P clarkii, so field applications must be carried out in the future if needed (in certain closed habitats) to identify a toxicant that exhibits rapid kill with minimum side effects on non-target organisms Appendix A Supplementary material Supplementary data associated with this article can be found, in the online version, at http://dx.doi.org/10.1016/j.jobaz.2016 08.001 References Alves, S.R.C., Patrı´ ci, C.S., Dominique, P.I., Angela, Z.D., Franklin, R.A., Luis, A.S., Alfonso, C.D., 2002 Effects of furadan in the brown mussel Perna perna and in the mangrove oyster Crassostrea rhizophorae Mar Environ Res 54, 241–245 Anasta´cio, P.M., Frias, A.F., Marques, J.C., 2000 Impact of crayfish densities on wet seeded rice and the inefficiency of a non-ionic surfactant as an ecotechnological solution Ecol Eng 15, 17–25 Anasta´cio, P.M., 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