Olivier_187-199.indd INTRODUCTION The utilization of freshwater fish to supplement pro- tein demand by humans, especially in rural areas, is a topic widely researched. Nevertheless, South Africa has always lagged behind in this effort when compared to densely populated countries such as China, Taiwan and Israel. Many reasons for this state of affairs can be given, like the customary sup- ply of marine fish from South Africa’s long shoreline with its rich, but lately diminishing, fish community. South Africa’s inland and rural communities have also not to a large extent developed an established appetite for freshwater fish. This may be due to the relatively effortless and uncomplicated accessibility to competitively priced alternative sources of pro- tein, such as chicken, small ruminants, pigs and cat- tle. The position may, however, change drastically. De- clining tendencies in marine fisheries output have been reported as long ago as the 1970s (Noble & Hemens 1978; Cram 1980; Allanson & Jackson 1983). Since then more documented evidence of this phenomenon is available. The imbalance be- tween the development of land protein resources on the one hand and population growth linked to rising costs of refrigeration and transport of marine protein products on the other, may, however, force the pen- dulum to move towards utilization of aquaculture products as supplementary food sources. This, to- gether with an increased interest in the develop- ment of aquaculture practices, has spurred the ini- tiation of research efforts in aspects such as intensive freshwater fish husbandry, harvesting of fish stocks from natural freshwater habitats (Roode 1978; Saay- 187 Onderstepoort Journal of Veterinary Research, 76:187–199 (2009) Report on some monogenean and clinostomid infestations of freshwater fish and waterbird hosts in Middle Letaba Dam, Limpopo Province, South Africa P.A.S. OLIVIER1*, WILMIEN J. LUUS-POWELL1 and J.E. SAAYMAN1, 2 ABSTRACT OLIVIER, P.A.S., LUUS-POWELL, WILMIEN J. & SAAYMAN, J.E. 2009. Report on some monoge- nean and clinostomid infestations of freshwater fish and waterbird hosts in Middle Letaba Dam, Limpopo Province, South Africa. Onderstepoort Journal of Veterinary Research, 76:187–199 This report deals with the results of a parasitological study done as part of a post-impoundment eco- logical study of Middle Letaba Dam, Limpopo Province, South Africa. It involved a seasonal survey protocol with particular attention to the diversity and prevalence of the parasitic fauna of the indige- nous fish community of the dam and the role of selected fish-eating birds in the life cycle and distribu- tion of fish helminths. The potential species composition of fish of the dam is provided. Monogenean and clinostomatid parasites encountered are listed and infestation statistics of fish and fish-eating bird hosts are presented. The results of this study also provide information on new distribution and host records of the encountered monogeneans. Keywords: Clinostomidae, Middle Letaba Dam, Monogenea, South Africa * Author to whom correspondence is to be directed: olivierp@ ul.ac.za 1 Department of Biodiversity, University of Limpopo (Turfloop Campus), Private Bag X1106, Sovenga, 0727 South Africa 2 Present address: 7 Protea Street, Albertinia, 6695 South Afri- ca Accepted for publication 1 September 2008—Editor 188 Monogenean and clinostomid infestations of freshwater fi sh and waterbird hosts in Limpopo, South Africa man 1984; Hecht 1985; Hecht, Uys & Britz 1988) and the enhancement of guidelines, aims and objec- tives for aquaculture-supportive research in South Africa (Safriel & Bruton 1984). Prioritizing freshwater fish as a supplementary alter- native supply of protein for humans indisputably ac- centuates the importance of fish health in general. In this regard, the significant role of scientific knowl- edge with regards to indigenous as well as intro- duced fish parasites can hardly be over-emphasized. Over the last decade the importance of studying fish parasites, including aspects such as their diversity, distribution, infestation rates and pathology have been realized by many South African researchers. This report deals with the results of parasitological studies done as part of a multidisciplinary research programme on the post-impoundment ecology of the Middle Letaba Dam. The study was commis- sioned in 1987 by the (then) Department of Devel op- ment Aid with the Government Service of the (then) Gazankulu Government as beneficiary. The study involved a seasonal survey protocol and particular attention was given to the diversity and prevalence of the parasitic fauna of the indigenous fish com- munity of the dam. Attention also focused on the role of selected waterbirds in the life cycle and dis- tribution of some fish helminths (with special refer- ence to members of the Clinostomidae) using pis- civorous birds as final hosts. MATERIALS AND METHODS Study area Middle Letaba Dam is situated at 30°24’ S, 23°16’ E, about 50 km west of the city of Giyani, in the Lim- popo Province of South Africa. The catchment area of the dam includes two river systems: the Sen wa- bathweni River with a catchment area of 757 km2 and a mean annual rainfall of 757 mm, and the Mid- dle Letaba River with a catchment area of 1 042 km2 and a mean annual rainfall of 708 mm. The most important tributaries of these systems are the Koe- does, Brandboontjies and Houtbos Rivers. At full water level, Middle Letaba Dam has a surface area of 1 943 ha and a mean depth of 9.5 m (maximum of 34 m). Water temperatures (day and night tem- peratures) and pH values were frequently meas- ured at various positions in the dam. Precipitation data were obtained from the Weather Bureau Sta- tion in the vicinity of the dam. Water temperatures did not fluctuate much seasonally, with average seasonal day temperatures being 27.5 °C (autumn), 17.5 °C (winter), 22 °C (spring) and 27 °C (summer) and average night temperatures being 26 °C (au- tumn), 16,5 °C (winter), 20 °C (spring) and 25.5 °C (summer). Reasons for these relatively small differ- ences may be due to the fact that the Middle Letaba Dam is situated in a subtropical environment and that at the time of the survey the dam was only about 30 % full. As a result, there was no clear delineation between the epilimnion and hypolimnion resulting in a thorough mixing of the entire water column. For the period of the survey pH values of the impound- ment were relatively constant, with average season- al values ranging between 7.9 (autumn), 8.3 (winter), 7.7 (spring) and 8.1 (summer). Procuring and transport of host specimens Parasitological surveys were done seasonally from January 1984 to January 1989. The majority of fish hosts were captured using gill nets of various stretched mess sizes, ranging from 30–180 mm but beach seine nets were used in shallower water. Where the physical conditions did not allow the use of gill nets fish hosts were collected by means of electro-fishing. The captured fish were transported live to the field laboratory where they were kept in containers with well aerated water. Avian hosts were shot and their beaks were immediately sealed with elastic bands to avoid the escape of any clinos- tomid worms lodged in the buccal cavity and oesophagus. The birds were transported in sealed plastic bags to the field laboratory for immediate au- topsy and parasitic infestation analysis of the com- plete alimentary tract. Examination of hosts for parasites Immediately prior to examination, the fish were killed by severing the spinal cord just posterior to the cranium. Monogenean parasites were collected from the gills of the fish with the aid of a stereo mi- croscope and smears were made from the skin and mucosa of the stomach. Sampled monogeneans were fixed in hot (70 °C) alcohol-formalin-acetic acid (AFA) and stored in 4 % buffered formalin. Some of the monogeneans were mounted on mi- croscope slides in glycerine jelly under slight cover slip pressure, and the cover slip sealed with clear nail varnish. Clinostomids were carefully removed from the buccal cavity and oesophagus of the birds, fixed in hot (70 °C) AFA, and preserved in 80 % ethanol. The clinostomids that were collected were mounted on microscope slides for microscopic in- vestigations or stored in 80 % ethanol for further studies. 189 P.A.S. OLIVIER, W.J. LUUS-POWELL & J.E. SAAYMAN Prevalence of infestation (%) was calculated as the number of infested hosts/number of examined hosts X 100. Mean intensity of infestation was calculated as the total number of parasites/number of infested hosts. Standard deviation was not calculated due to meaningless values as a result of too small sample sizes (Rózsa, Reiczigel & Majoros 2000). For con- sistency, name changes of hosts since the begin- ning of this study are incorporated, following that of Skelton (2001) for fish hosts and Hockey, Dean & Ryan (2005) for bird hosts. RESULTS AND DISCUSSION Fish species composition of Middle Letaba Dam A pilot-survey (done prior to the parasitological sur- vey) of the catchment area of the proposed Middle Letaba Dam was carried out to determine the pos- sible species composition of fish which could even- tually inhabit the dam (Olivier, Kruger, Van der Waal, Viljoen & Viljoen 1986). The latter investigation com- plements a previous study by Gaigher & McPott (1973), and mainly correlates with their findings, ex- cept for Barbus radiatus Peters and Synodontis zam bezensis Peters which the latter authors en- countered in the Middle Letaba River and Glosso- gobius giuris (Hamilton-Buchanan) which they col- lected at the confluence of the Middle Letaba and Klein Letaba Rivers. Based on the results of these two surveys, the potential fish species diversity of the dam is relatively low, limited to about 28 species with Clarias gariepinus Burchell and Oreochromis mossambicus (Peters) the numerically dominant species (Table 1). Of the species mentioned in Table 1, Mesobola bre- vianalis (Boulenger), Opsaridium zambezense (Gil- christ et Thompson), Barbus lineomaculatus Bou- lenger, B. radiatus, Cyprinus carpio L., Micralestes acutidens (Peters), Amphilius uranoscopus (Pfef fer), Schilbe intermedius Rüppell, Chiloglanis pretoriae Van der Horst, S. zambezensis, Micropterus salmoi- des (Lacepède), Tilapia rendalli (Boulenger) and G. giuris were not collected from the dam during the present parasitological survey. Some of these may TABLE 1 Potential species composition of fish of Middle Letaba Dam and its catchment area Gaigher & McPott 1973 n Olivier et al. 1986 Sampled during this study Anguilla mossambica Peters Mesobola brevianalis (Boulenger) Opsaridium peringueyi (Gilchrist & Thompson) Barbus lineomaculatus Boulenger Barbus unitaeniatus Günther Barbus bifrenatus Fowler Barbus viviparus Weber Barbus toppini Boulenger Barbus radiatus Peters Barbus trimaculatus Peters Barbus paludinosus Peters Labeobarbus marequensis (A. Smith) Labeo rosae Steindachner Labeo ruddi Boulenger Labeo cylindricus Peters Labeo molybdinus Du Plessis Cyprinus carpio Linnaeus Micralestes acutidens (Peters) Amphilius uranoscopus (Pfeffer) Schilbe intermedius (Rüppell) Clarias gariepinus (Burchell) Chiloglanis pretoriae Van der Horst Synodontis zambezensis Peters Micropterus salmoides (Lacepède) Pseudocrenilabrus philander (Weber) Tilapia rendalli (Boulenger) Oreochromis mossambicus (Peters) Glossogobius giuris (Hamilton-Buchanan) 1 – – 82 13 46 68 – 89 83 32 24 64 68 4 – – – – 111 – – – 69 – 177 – + + + + + + + + X + + + + + + + + + + + + + X + + + + X + X X X + + + + X + + + + + + + X X X X + X X X + X + X n = number of fish sampled for parasitological analyses + = species present X = species not collected 190 Monogenean and clinostomid infestations of freshwater fi sh and waterbird hosts in Limpopo, South Africa still successfully colonize Middle Letaba Dam. How- ever, C. pretoriae and O. zambezense are consid- ered to prefer rivers as their favoured habitat and will, in all probability, not establish themselves in the dam (Polling, Mokgalong & Saayman 1983). Syno- dontis zambezensis and G. giuris might also not colonize the dam as they naturally occur only in downstream habitats (Gaigher 1973). The unique construction of the overflow of the dam, forming an impenetrable barrier to upstream migration, will pre- vent these two species from reaching the dam. Of the 28 species of fish listed in Table 1, only 15 were collected during the parasitological survey and, therefore, could be considered as possible parasite hosts (only a single specimen of Anguilla mossam- bica Peters was collected during the summer sur- vey of 1984, but revealed no parasites). Monogenean parasites To date, very few records of Monogenea of South African freshwater fish appeared in the published literature (e.g. Paperna 1980; Mashego 1983; Khalil & Polling 1997; Mashego 2000; Luus-Powell, Ma- shego & Khalil 2003; Christison, Shinn & Van As 2005). During this study monogenean parasites were collected from a total of 831 hosts specimens of as many as 14 different fish species (Table 2). TABLE 2 Host/parasite checklist of monogenean parasites from Middle Letaba Dam with an indication of host specificity for each species of parasite Host n Parasite Host range Barbus unitaeniatus Günther 73 Dactylogyrus sp. 1 3 Barbus bifrenatus Fowler 13 Dactylogyrus sp. 4 3 Barbus viviparous Weber 46 Dactylogyrus sp. 4 3 Barbus toppini Boulenger 68 Dactylogyrus sp. 4 3 Barbus trimaculatus Peters 70 Dactylogyrus afrolongicornis Paperna Dactylogyrus afrolongicornis alberti Paperna Dactylogyrus afrosclerovaginatus Paperna Dactylogyrus allolongionchus Paperna Dactylogyrus meyersi Price, McClellan, Druckenmiller & Jacobs Dactylogyrus sp. 1 Dactylogyrus sp. 2 Dactylogyrus sp. 3 1 1 1 1 1 3 2 1 Barbus paludinosus Peters 83 Dactylogyrus dominici Mashego Dactylogyrus teresae Mashego 1 1 Labeobarbus mareqensis (A. Smith) 32 Dactylogyrus spinicirrus (Paperna & Thurston) Dactylogyrus sp. 1 Dactylogyrus sp. 2 1 3 2 Labeo rosae Steindacher 24 Dactylogyrus sp. 7 Dactylogyrus sp. 8 Dactylogyrus sp. 9 Dactylogyrus sp. 10 Dogielius sp. 1 2 2 2 2 1 Labeo ruddi Boulenger 64 Dactylogyrus sp. 7 Dactylogyrus sp. 9 Dactylogyrus sp. 9 Dactylogyrus sp. J Dactylogyrus sp. 10 Dactylogyrus sp. 12 Dogielius sp. 2 Dogielius sp. 3 Dogielius sp. 4 2 2 2 2 1 1 1 1 1 Labeo cylindricus Peters 68 Dactylogyrus sp. 5 2 Labeo molybdinus Du Plessis 4 Dactylogyrus sp. 5 Dactylogyrus sp. 6 2 1 191 P.A.S. OLIVIER, W.J. LUUS-POWELL & J.E. SAAYMAN Host n Parasite Host range Clarias gariepinus (Burchell) 111 Quadriacanthus allobychowskiella Paperna Quadriacanthus aegypticus El-Naggar & Serag Quadriacanthus clariadis Paperna Quadriacanthus sp. 1 Quadriacanthus sp. 2 Quadriacanthus sp. 3 Quadriacanthus sp. 4 Quadriacanthus sp. 5 Quadriacanthus sp. 6 Quadriacanthus sp. 7 Quadriacanthus sp. 8 Quadriacanthus sp. 9 Quadriacanthus sp. 10 Gyrodactylus rysavyi Ergens Macrogyrodactylus clarrii Gussev Macrogyrodactylus karibae Douellou & Chishawa 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Pseudocrenilabrus philander (Weber) 69 Cichlidogyrus tilapiae Enterogyrus cichlidarium 2 2 Oreochromis mossambicus (Peters) 105 Cichlidogyrus halli (Price & Kirk) Cichlidogyrus sclerosus Paperna & Thurston Cichlidogyrus tilapiae Paperna Cichlidogyrus sp. 1 Cichlidogyrus sp. 2 Enterogyrus cichlidarum Paperna Scutogyrus gravivaginus (Paperna & Thurston) 1 1 2 1 1 2 1 n = number of hosts examined 1 = one-host parasite 2 = two-host parasite 3 = three-host parasite TABLE 2 (cont.) Although some of the parasites encountered are of uncertain specific status, their generic status and host distribution were positively verified. The dactylogyrid species were obtained from the gills of their respective hosts. Gyrodactylid parasites were found to be present on either the skin or the gills of the host. Species of Chichlidogyrus Paperna and Scutogyrus Pariselle & Euzet are harboured on the gills of their hosts, while Enterogyrus chichl- idarum Paperna occurs in the mucosa of the stom- ach of both its hosts (Pseudocrenilabrus philander (Weber) and O. mossambicus). During the current study, members of three mono- genean families, represented by eight genera and 47 species (19 known species and 28 unidentified species), were encountered (Table 2). Species of these families are known to be variably host specif- ic, with host ranges restricted to one or only a few closely related species. This trend is supported by the results of this study (Table 2). In general, mem- bers of the genus Dogielius Bikhovski were found only on two of the four Labeo hosts and species of Quadriacanthus Paperna, Gyrodactylus Paperna and Macrogyrodactylus Malberg were found on C. gariepinus only. Species of Cichlidogyrus were re- stricted to P. philander and O. mossambicus, while Scutogyrus gravivaginus (Paperna & Thurston) was collected from O. mossambicus only. Species of the genus Dactylogyrus Diesing are parasites of Barbus and Labeo species. More specifically, Table 2 clear- ly illustrates the phenomenon of host specificity with no less than 37 of encountered species found on a single host, eight parasitized two-host species and only two were harboured by three different hosts. This corresponds with findings of Mashego (1983) who reported on eight of the parasites collected dur- ing the present study parasitizing the same host species, although from different geographical re- gions in South Africa. These are Dactylogyrus afro- longicornis afrolongicornis Paperna, Dactylogyrus afrolongicornis alberti Paperna, Dactylogyrus afro- sclerovaginatus Paperna, Dactylogyrus allolongion- chus Paperna and Dactylogyrus meyersi Price, Mclellan, Druckenmiller & Jacobs from Barbus tri- maculatus Peters, Dactylogyrus dominici Mashego 192 Monogenean and clinostomid infestations of freshwater fi sh and waterbird hosts in Limpopo, South Africa and Dactylogyrus teresae Mashego from Barbus pa ludinosus Peters, and Dactylogyrus spinicirrus (Paperna & Thurston) from Labeobarbus marequen- sis (Smith, 1841). In general, the intensity of monogenean infestations was found to be very low, on average five parasites per infested host. However, the diversity of mono- geneans on C. gariepinus (with no less than 16 spe- cies), with a more profound and variable intensity of infestation, is noteworthy. Immediately after the clo- sure of the dam in 1984, very low intensities were observed (mean intensity = 1.0) followed by a tre- mendous increase, reaching a peak in the summer of 1986/1987 (mean intensity = 700.0). This was fol- lowed by a steady decline to as low as a mean in- tensity of less than 1.0 in January 1988. The latter could be ascribed to the greater volume of water as a result of good rains in December/January of 1987/ 1988 (more than 250 mm; mean annual for the catchment area is 732 mm) resulting in a decline in host density. Gyrodactylids have no free swimming larval stages and infestation between individual hosts is probably by direct contact (Paperna 1980). During the period of hyperinfestation, infested spec- imens of C. gariepinus all developed a thick layer of greyish-white mucus over the entire skin, in all prob- ability caused by the severe irritation caused by the parasites. Other pathological signs observed in- cluded proliferation of epithelial cells at the point of attachment, erosion of the skin and fading of skin colour, all of which are in accord with similar obser- vations reported by Paperna (1980). Monogeneans encountered from Middle Letaba Dam revealed some species representing either first parasite records, or new host records from South Africa. This can be summarized as follows: 1. Gyrodactylus Von Nordman (Gyrodactylidae). Christison et al. (2005) provides a summary of all the gyrodactylids that have been reported from southern Africa. These include one known, Gyrodactylus transvaalensis Prudhoe & Hussey, and three unidentified species as records from South Africa. None of the latter was collected during this survey. However, the authors agree with Christison et al. (2005) that the records of the three unidentified species referred to by Bragg (1991) and Lombard (1968), should not be considered as part of the South African fauna due to their association with alien fish hosts (rainbow trout and large mouth black bass, re- spectively). However, Gyrodactylus rysavyi Er- gens, described and reported to date only from Egypt (Ergens 1973), was encountered during the present study from C. gariepinus, and repre- sents a new distribution record from South Afri- ca. 2. Macrogyrodactylus Malberg (Gyrodactylidae). During this study two macrogyrodactylid species (Macrogyrodactylus clarii Gussev and Macro- gyrodactylus karibae Douëllou & Chishawa) were encountered from C. gariepinus and have previ- ously been reported on by Khalil & Mashego (1998). The latter authors also reported on Mac- rogyrodactylus congolensis (Prudhoe) collected from C. gariepinus at Mokgoma-Matlala Dam, South Africa. 3. Dactylogyrus Diesing (Dactylogyridae). Eleven dactylogyrid species were previously reported from South Africa (Khalil & Polling 1997): D. af- ro longicornis Paperna, D. afrolongicornis alberti, D. afrosclerovaginus, D. allolongionchus, D. do- minici, Dactylogyrus enidae Mashego, Dac tyl o- gyrus jubbstrema Price, Korach & McPott, D. myersi, Dactylogyrus pienaari Price, Korach & McPott and D. spinicirrus D. teresae Mashego. Of these, only three species, D. enidae, D. jubb- strema and D. pienaari, were not encountered during this study. Of the remaining seven spe- cies collected, it is worthwhile to note that D. afrosclerovaginus was collected from Barbus trimaculatus Peters (previously reported from Barbus paludinosus Peters), representing a new host record. Additional to the above, 12 unidenti- fied dactylogyrid species were found from six hosts (Table 2). 4. Dogielius Bikhovski (Dactylogyridae). None of the more than 20 described species from Africa (Khalil & Polling 1997) was either sampled dur- ing the present study or is previously known from South Africa. However, four unidentified spe- cies, one from Labeo rosae Steindachner and three from Labeo ruddi Boulenger, were collect- ed. 5. Quadriacanthus Paperna (Dactylogyridae). Three formerly described species (Quadriacanthus ae- gypticus El-Naggar & Serag, Quadriacanthus al- lobychowskilla Paperna and Quadriacanthus clariadis Paperna) and ten unidentified species, all sampled from C. gariepinus and all being first records for the genus from South Africa, were collected. 6. Chichlidogyrus Paperna (Ancyrocephalidae). Only one species, Chichlidogyrus papernastrema Price, Peebles & Bamford is previously known 193 P.A.S. OLIVIER, W.J. LUUS-POWELL & J.E. SAAYMAN from South Africa. This species was not collect- ed during the present survey, possibly due to the absence of the type host, Tilapia sparrmanii Boulenger. However, five chichlidogyrid species were encountered and are new records for South Africa: Chichlidogyrus halli (Price & Kirk), Chi ch- lidogyrus sclerosus Paperna & Thurston, Chichli- dogyrus tilapiae Paperna and two unidentified species. 7. Enterogyrus Paperna (Ancyrocephalidae). En ter- o gyrus cichlidarum Paperna was the only spe- cies collected and represents a first record of the genus from South Africa. 8. Scutogyrus Pariselle & Euzet (Ancyrocephalidae). Scutogyrus gravivaginus (Paperna & Thurston) was the only species collected and it is a first record of the genus from South Africa. Digenean species of the family Clinostomidae Positive species designation of trematodan meta- cercariae is usually difficult due to their undifferenti- ated genitalia, therefore adult stages from the de- finitive host are frequently the only way to achieve positive identification. However, the fact that most trematodes, including their larval stages, are highly host specific (Paperna 1980; Grobler & Mokgalong 2002) can be applied as a useful tool for identifica- tion. Furthermore, diagnosis during this survey was somewhat easier as trematode infestations (espe- cially that of clinostomids) of resident piscivorous birds acting as definitive hosts, were simultaneously investigated (Mokgalong 1996). In the past, many problems were experienced with the specific diagnostic designations of members of the family Clinostomidae (Grobler, Mokgalong & Saayman 1999). Feizullaev & Mirzoeva (1983) pro- vided some clarity on solving this problem when they synonomized 35 species as one, Clinostomum complanatum (Rudolphi). These included Clino sto- mum vanderhorsti Ortlepp and Clinostomum tila- piae Ukoli, both of which prompted much discus- sion, confusion and misidentification. Recent studies using molecular techniques clearly revealed that material previously identified as C. tilapiae, found in the gill chambers of O. mossambicus, is in fact Neutraclinostomum intermedialis (Lamont) (Grobler et al. 1999; Grobler & Mokgalong 2002). The dem- onstration of high host specificity (O. mossambicus as intermediate fish host and the darter, Anhinga rufa (Daudin), as definitive avian host) for N. inter- medialis (Mokgalong 1996) further supports the view of considering C. tilapiae sampled from gill chambers of O. mossambicus, as a junior synonym of N. intermedialis. This approach is followed for the present report. During the present survey three species of the fam- ily Clinostomidae were sampled from six different fish hosts (Table 3). The results concerning these species are discussed separately below and, where applicable, additional data of similar studies from different habitats in South Africa are presented as comparisons. Neutraclinostomum intermedialis (Lamont) This parasite was exclusively procured from the branchial chambers of O. mossambicus and, during this study, a total of 379 specimens were collected. A total of 177 fish hosts were examined and showed a prevalence of 48 % and a mean intensity of 4.4 (Table 3). The largest number of hosts were en- countered during summer (n = 87), with almost 50 % (n = 39) represented by smaller fish (total length < 10 cm). Fish longer than 30 cm were virtually ab- sent from this survey, but the data of a single large specimen is nevertheless included in Table 4. TABLE 3 Species of the family Clinostomidae collected from infested intermediate hosts from Middle Letaba Dam Parasite Host n Prev. Mi. Neutraclinostomum intermedialis (Lamont) Oreochromis mossambicus (Peters) 177 48 4.4 Clinostomum complanatum (Rudolphi) Barbus unitaeniatus Günther Barbus trimaculatus Peters Labeobarbus marequensis (A. Smith) Pseudocrenilabrus philander (Weber) 82 89 13 47 2.4 1.1 7.7 6.4 1 2 1 1 Euclinostomum heterostomum (Rudolphi) Oreochromis mossambicus (Peters) Clarias gariepinus (Burchell) 177 21 24.9 4.7 2.2 9 n = number of hosts examined Prev. = prevalence of infestation (%) Mi. = mean intensity of infestation 194 Monogenean and clinostomid infestations of freshwater fi sh and waterbird hosts in Limpopo, South Africa The highest prevalence (57.1 % for all seasons) was found within the < 10 cm length group with a 63.6 % registered for the winter survey within the same length group (the autumn survey was not tak- en into account due the fact that only two hosts were examined). The lowest prevalence was found for hosts of 20.1–30 cm in length suggesting small- er fish to be more susceptible to infestation. The highest mean intensity (11.3), however, was record- ed during the summer survey for the 20.1–30 cm length group with as many as 20 parasites per host. This length group in fact recorded the highest mean intensity values for all seasonal surveys, except during spring (Table 4). Neutraclinostomum intermedialis does indicate some, yet not pronounced, seasonal variations in prevalence and intensity of infestation. This rela- tively small seasonal variation in infestation rates is to be expected in a young, subtropical impound- ment where seasonal climatic conditions are not sharply demarcated. Furthermore, comparison of infestation rates amongst different sexes of the host failed to illustrate any noteworthy differences. Mokgalong (1996) clearly illustrated A. rufa to be the definitive host for N. intermedialis, with 75 % of the examined hosts from Middle Letaba Dam infest- ed (Table 5). He further reported high infestation values of N. intermedialis for this bird (intensity = 7–72 and mean intensity = 35). These values cor- respond well (prevalence of 50–100 %) if compared with similar data for other water bodies in the Limpopo Province (Mokgalong 1996). The high in- festation values of A. rufa in Middle Letaba Dam, and the fact that it is one of the most abundant resi- TABLE 4 Seasonal analysis, per length group (TL. cm) of Oreochromis mossambicus (Peters), for Neutraclinostomum intermedialis (Lamont) infestations in Middle Letaba Dam Season < 10 cm 10.1–20 cm 20.1–30 cm > 30 cm All size groups Summer Number of hosts examined Prevalence of infestation Intensity of infestation Mean intensity of infestation Number of parasites collected 39 53.9 1–21 4 83 24 45.8 1–7 4.3 47 23 13.1 5–20 11.3 34 1 100 4 4 4 87 41.4 1–21 4.7 168 Autumn Number of hosts examined Prevalence of infestation Intensity of infestation Mean intensity of infestation Number of parasites collected 2 100 3–4 3.5 7 10 80 2–16 5.6 45 3 33.3 – 8 8 – – – – – 15 73.3 2–16 5.5 60 Winter Number of hosts examined Prevalence of infestation Intensity of infestation Mean intensity of infestation Number of parasites collected 11 63.6 1–5 2.4 17 4 75 4–5 4.3 13 25 40 1–17 3.7 37 – – – – – 40 50 1–17 3.4 67 Spring Number of hosts examined Prevalence of infestation Intensity of infestation Mean intensity of infestation Number of parasites collected 18 55.6 1–10 5.5 55 14 50 1–9 4 28 3 33.3 – 1 1 – – – – – 35 51.4 1–10 4.7 84 All seasons Number of hosts examined Prevalence of infestation Intensity of infestation Mean intensity of infestation Number of parasites collected 70 57.1 1–21 4.1 16 52 55.8 1–16 4.6 133 54 27.8 1–20 5.6 80 1 100 4 4 4 177 48 1–21 4.5 379 195 P.A.S. OLIVIER, W.J. LUUS-POWELL & J.E. SAAYMAN dent fish-eating birds (Olivier, Saayman & Polling 1991), tend to indicate that intensity of infestation in the intermediate host (O. mossambicus) may well increase drastically in the near future. Saayman (1986) investigated N. intermedialis infes- tations in O. mossambicus in various other water bodies of the Limpopo and Olifants Rivers Drainage Systems in the Limpopo Province (Table 6). Only three of these water bodies (Nile Flood Pans, Nwanedzi River and Tompi Seleka Fish Station) ex- hibit infestation values comparable to that of Middle Letaba Dam. Values obtained from Tompi Seleka are to be expected as it represents a situation where fish are kept at high densities in production ponds. These ponds were also densely populated with snails, the invertebrate intermediate host for N. in- termedialis, (no data on the species of the snails are available) and aquatic macrophytes. The ratio of shore to water area in production ponds is far great- er than that for impoundments, resulting in en- hanced opportunities for released cercariae to lo- cate suitable hosts. Physical conditions in the Nile Flood Pans and pools in the Nwanedzi River are almost similar to the situation at Tompi Seleka and this may explain the high levels of infestation re- corded for these two localities. Clinostomum complanatum (Rudolphi) In Middle Letaba Dam only four of the potential fish host species (Table 1) were infested with metacer- TABLE 5 Parasite/host checklist of definitive hosts (piscivorous birds) at Middle Letaba Dam infested with adult members of Clinostomidae Parasite Host n Prev. In. Mi. Neutraclinostomum intermedialis (Lamont) Anhinga rufa (Daudin) 44 75 7–72 35 Clinostomum complanatum (Rudolphi) Phalacrocorax lucidus (Lichtenstein) Phalacrocorax africanus (Gmelin) Anhinga rufa (Daudin) Ardea cinerea Linnaeus Nycticorax nycticorax (Linnaeus) 11 30 44 3 3 63 60 96 66 66 1–37 1–7 3–190 1–3 2–4 11 4 48 2 3 Euclinostomum heterostomum (Rudolphi) Phalacrocorax luidus (Lichtenstein) Phalacrocorax africanus Anhinga rufa (Daudin) Ardea cinerea Linnaeus Ardea melanocephala Anon Ardea purpurea Linnaeus 11 30 44 3 5 2 88 30 80 100 100 50 1–21 1–10 1–11 4–6 1–8 1 7 6 5 5 5 1 n = number of hosts examined Prev. = prevalence of infestation (%) In. = intensity of infestation Mi. = mean intensity of infestation TABLE 6 Comparison of infestation values of Neutraclinostomum intermedialis (Lamont) in Oreochromis mossambicus (Peters) in different water bodies of the Limpopo and Olifants Rivers Drainage Systems—data from Saayman (1986) Waterbody n Prev. In. Mi. Middle Letaba Dam Glen Alpine Dam Luphephe-Nwanedzi Dams Nwanedzi River Nzhelele Dam Seshego Dam Nile Flood Pans Piet Gouws Dam Lepellane Dam Tompi Seleka Fish Station 177 89 157 66 15 35 58 59 114 25 48 14.6 10.7 54.6 7 38 62.1 35.6 28.9 64 1–21 1–6 1–5 1–8 1 1–5 1–13 1–14 1–6 1–2 4.5 2 1.4 3.2 1 3 3.5 2.8 2.1 1.5 n = number of hosts examined Prev. = prevalence of infestation (%) In. = intensity of infestation Mi. = mean intensity of infestation 196 Monogenean and clinostomid infestations of freshwater fi sh and waterbird hosts in Limpopo, South Africa cariae of C. complanatum (Table 3). In all of the in- fested hosts the parasites were always found en- cysted on the peritoneum of visceral organs with an explicit preference for the peritoneum of the ventral surface of the swim bladder. Extremely low preva- lence and intensity values were recorded (Table 3). Saayman (1986) investigated several indigenous fish species from various localities in the Olifants and Limpopo Drainage Systems for C. complana- tum infestations. From the numerous hosts investi- gated, only five species (including Marcusenius macrolepidotus (Peters) which was not encoun- tered in Middle Letaba Dam) were infested (Table 7). From the infested hosts, a total of 1985 parasites were procured with prevalence and intensity values, as given in Table 7, profoundly higher than those recorded for Middle Letaba Dam. Seven piscivorous birds at Middle Letaba Dam were identified by Mokgalong (1996) as regular final hosts for C. complanatum (Table 5). Although relatively low numbers of birds were examined, the data of Table 5 clearly indicate A. rufa as the predominant definitive host for this parasite. Reed cormorants, Phalacrocorax africanus (Gmelin), grey herons, Ardea cinerea Linnaeus. and black-crowned night herons, Nycticorax nycticorax Linnaeus, seem to act as subsidiary final hosts. If the results from the present study and that of Saayman (1986) are evaluated, the following impor- tant observation needs to be elucidated: S. interme- dius and M. macrolepidotus seem to be the major intermediate hosts for C. complanatum (Table 7). Both were, however, absent from Middle Letaba Dam at the time of the survey, but S. intermedius does occur in the catchment area of the dam (Table 1). It can, therefore, be expected that the latter will colonize the dam at some stage. As the biological requirements for the completion of the life cycle of the parasite are established in the dam, it could well be expected that S. intermedius would immediately become infested after colonization. Euclinostomum heterostomum (Rudolphi) During this study, E. heterostomum was recorded regularly, encysted in muscle tissue of the host. The major intermediate host was identified as O. mos- sambicus from which a total of 96 parasites were collected (Table 8). A single male C. gariepinus (to- TABLE 7 Fish intermediate hosts from various localities in the Olifants (O) and Limpopo (L) Rivers Drainage Systems infested with Clinostomum complanatum (Rudolphi)—data from Saayman (1986) Host n Prev. In. Mi. Locality and drainage system Schilbe intermedius (Rüppell) 399 28.7 1–21 3.4 Luphephe-Nwanedzi Dams (L) Nwanedzi River (L) Glen Alpine Dam (L) Magalakwena River (L) Nzhelele Dam (L) Nile Flood Pans (L) Ga-Selati River (O) Olifants River (O) Letaba Estates (O) Marcusenius macrolepidotus (Peters) 524 54.7 1–122 6.7 Luphephe-Nwanedzi Dams (L) Nwanedzi River (L) Glen Alpine Dam (L) Nzhelele Dam (L) Nile Flood Pans (L) Mohlapitse River (O) Letaba Estates (O) Barbus unitaeniatus Günther 2 50 1 1.0 Letaba Estates (O) Barbus toppini Weber 1 100 8 8.0 Letaba Estates (O) Chiloglanis pretoriae Van der Horst 115 5.2 1–4 2.2 Luvhuvhu River (L) Nwanedzi River (L) Mohlapitse River (O) n = number of hosts examined Prev. = prevalence of infestation (%) In. = intensity of infestation Mi. = mean intensity of infestation 197 P.A.S. OLIVIER, W.J. LUUS-POWELL & J.E. SAAYMAN tal length = 66,3 cm), from a sample of 21 speci- mens examined, collected during the winter survey, was found harbouring nine specimens of E. heter- ostomum. This may suggest that C. gariepinus, in extreme cases, can act as a reservoir host. The data of Table 8 reflect that all length groups of O. mossambicus may become infested with the par- asite although it appears as if larger fish (total length longer than 20 cm) may be more vulnerable to in- festation. No significant seasonal variations in infes- tation, nor any prevalence amongst different sexes, were observed. In Middle Letaba Dam it was established that mem- bers of the Ardeidae, Phalacrocoracidae and Anhingidae act as final hosts for the adults of E. het- erostomum (Table 5), the latter firmly attached to the lining of the buccal cavity and oesophagus. Except for P. africanus with a prevalence of 30 %, all the other hosts demonstrated higher prevalence values ranging from 50–100 % with intensity values ranging from 1–21. White-breasted cormorants, Phalacrocorax lucidus (Lichtenstein), and A. rufa seem to be the major definitive hosts, but, A. cine- rea, black-headed herons, Ardea melanocephala (Anon), and purple herons, Ardea purpurea Lin- naeus, may well prove to be equally or even more important, pending the examination of larger num- bers of hosts. Although clinostomid metacercarial cysts attain a relatively large size, and notwithstanding the rela- tively high infestation values, it does not appear as if these parasites cause any deleterious affects to semi-adult and/or adult hosts. This observation is supported by Paperna (1980). In fingerlings and TABLE 8 Seasonal analysis per length group (TL, cm) of Oreochromis mossambicus (Peters) for Euclinostomum heterostomum (Rudolphi) infestations in Middle Letaba Dam Season < 10 cm 10.1–20 cm 20.1–30 cm > 30 cm All size groups Summer Number of hosts examined Prevalence of infestation Intensity of infestation Mean intensity of infestation Number of parasites collected 39 12.8 1 1 5 24 33.3 1–9 2.6 21 23 56.5 1–8 2.2 29 1 100 5 5 5 87 31 1–9 2.2 60 Autumn Number of hosts examined Prevalence of infestation Intensity of infestation Mean intensity of infestation Number of parasites collected 2 – – – – 10 20 1–2 1.5 3 3 – – – – – – – – – 15 13.3 1–4 1.5 3 Winter Number of hosts examined Prevalence of infestation Intensity of infestation Mean intensity of infestation Number of parasites collected 11 9.1 1 1 1 4 – – – – 25 36 1–4 2.1 19 – – – – – 40 25 1–4 2 20 Spring Number of hosts examined Prevalence of infestation Intensity of infestation Mean intensity of infestation Number of parasites collected 18 5.6 1 1 1 14 28.6 1–6 3 12 3 – – – – – – – – – 35 14.3 1–6 2.6 13 All seasons Number of hosts examined Prevalence of infestation Intensity of infestation Mean intensity of infestation Number of parasites collected 70 10 1 1 7 52 26.9 1–9 2.6 36 54 43.1 1–8 2.2 48 1 25 1 1 5 177 24.9 1–9 2.2 96 198 Monogenean and clinostomid infestations of freshwater fi sh and waterbird hosts in Limpopo, South Africa smaller fish species, however, severe infestations are likely to be detrimental. Paperna (1980) report- ed mortalities amongst very young (40–60 mm) O. mossambicus, even with intensity values of as low as 3–5 worms. During the present survey a number of juvenile specimens of O. mossambicus infested with E. heterostomum showed definite signs of lo- comotory impairment. Severe infestation with N. in- termedialis is also likely to cause respiratory inhibi- tion. Britz, Saayman & Van As (1984) and Britz, Van As & Saayman (1984) reported considerable physical and histological damage to the oesophageal muco- sa of avian hosts infested with clinostomid worms. This supports earlier reports from Baugh & Pandey (1969) and Ukoli (1970). The latter author observed that if worms are removed from the oesophageal region of the host, part of the host tissue is torn away, leaving a deep lesion which bleeds freely. The study at Middle Letaba Dam confirmed these observations in almost all infested avian hosts. ACKNOWLEDGEMENTS The authors thank the Department of Development Aid, Premier Food Industries Ltd. and the Research Development and Administration of the University of Limpopo for financial assistance. We also thank the (then) Gazankulu Government for their involve- ment and support and the Department of Biodiversity, University of Limpopo for infrastructure and techni- cal support. REFERENCES ALLANSON, R.B. & JACKSON, P.B.N. 1983. Limnology and fisheries potential of Lake PK le Roux. Pretoria: CSIR (South African National Scientific Programmes. Report no. 77). BAUGH, C.H. & PANDEY, K.C. 1969. Studies on clinostome metacercariae. I. A restudy of Clinostomum giganticum. An- gewandte Parasitologie, 10:211–223. BRAGG, R.R. 1991. Health status of salmonids in river systems in Natal. I. Collection of fish and parasitological examination. Onderstepoort Journal of Veterinary Research, 58:59–62. BRITZ, J., SAAYMAN, J.E. & VAN AS, J.G. 1984. 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