Austin_227-234.indd INTRODUCTION In the genus Lamproglena Von Nordmann, 1832 only the adult females are gill parasites of fishes. The majority of species occur in freshwater and the only exception, Lamproglena lichiae Von Nordmann, 1832 was collected from the Red Sea (Fryer 1968) on the doublespotted queenfish, Scomberoides ly- san. Six of the 39 species of this genus recorded are from southern Africa (Moll & Avenant-Oldewage 2008) and include Lamproglena monodi Capart, 1944; Lamproglena clariae Fryer, 1956; Lampro- glena barbicola Fryer, 1961; Lamproglena cornuta Fryer, 1964; Lamproglena hoi Dippenaar, Luus- 227 Onderstepoort Journal of Veterinary Research, 76:227–234 (2009) Ecological parameters of Lamproglena hoi (Copepoda: Lernaeidae) infection on the Bushveld smallscale yellowfish, Labeobarbus polylepis (Boulenger, 1907) A. AUSTIN and A. AVENANT-OLDEWAGE* Department of Zoology, University of Johannesburg, Kingsway Campus, P.O. Box 524, Auckland Park, Johannesburg, 2006 South Africa * E-mail address: aoldewage@uj.ac.za Accepted for publication 12 November 2008—Editor ABSTRACT AUSTIN, A. & AVENANT-OLDEWAGE, A. 2009. Ecological parameters of Lamproglena hoi (Cope- poda: Lernaeidae) infection on the Bushveld smallscale yellowfish, Labeobarbus polylepis (Bou- lenger, 1907). Onderstepoort Journal of Veterinary Research,76:227–234 This study describes the distribution and aspects of the ecology of Lamproglena hoi. Bushveld small- scale yellowfish, Labeobarbus polylepis (Boulenger, 1907) were collected during June 2006 from the Phongolo and Assegaai rivers, March 2005 and October 2006 from the Elands River, and January 2007 and June 2008 from the Komati River in Mpumalanga, South Africa and examined for the pres- ence of parasites. Lamproglena hoi specimens were collected from the gill filaments of the host. Specimens were fixed with warm AFA (alcohol-formaldehyde-acetic acid) and preserved in 70 % ethanol. The identification of parasites took place in the laboratories of the University of Johannesburg. Twenty-five copepods (prevalence 21 %, mean intensity = 4.17, abundance = 0.86) were collected on 29 fish in the Phongolo River and 46 copepods (prevalence 40 %, mean intensity = 3.83, abundance = 1.53) were collected on 30 fish in the Assegaai River. One hundred and sixty eight copepods (prevalence 52 %, mean intensity = 12.92, abundance = 6.72) were collected on 25 fish in 2005, and 527 copepods (prevalence 95 %, mean intensity = 27.74, abundance = 26.35) were collected on 20 fish in the Elands River. One hundred and sixteen copepods (prevalence 75 %, mean intensity = 7.73, abundance = 5.80) were collected on 20 fish in 2007, and 273 copepods (prevalence 63 %, mean intensity = 16.06, abundance = 10.11) were collected on 27 fish in 2008 in the Komati River. Labe o- barbus polylepis from these four rivers was found to have a relatively high L. hoi infection. Inseminated L. hoi females (immature) attach to the host in winter and their ovaries become con- spicuous (mature). In spring fertilized eggs are stored in egg sacs hanging from the body (gravid), indicating that fertilized eggs start to hatch in spring and continued hatching into summer. Parasites prefer the median part of the second gill arch for attachment. No correlation exists between the number of parasites recorded on the gills and the sizes (total lengths) of yellowfish sampled. Keywords: Copepoda, Crustacea, ecology, Labeobarbus polylepis, Lamproglena hoi, yellowfish 228 Lamproglena hoi infection on Bushveld Smallscale Yellowfi sh, Labeobarbus polylepis (Boulenger, 1907) Powell & Roux, 2001; Lamproglena hepsetii Van As & Van As, 2006. Lamproglena hoi was described from specimens col- lected in the Spekboom River, Mpumalanga (Dip pe- naar, Luus-Powell & Roux 2001). It is an ectopara- site on the gills of the Lowveld largescale yellowfish, Labeobarbus marequensis and the Bush veld small- scale yellowfish, Labeobarbus polylepis. Labeobar- bus polylepis is an indigenous species endemic in the southern tributaries of the Limpopo River System, as well as the Inkomati and Phongolo sys- tem, above altitudes of 600 m and prefers perma- nent rivers with deep pools, riffles and runs (Skelton 2001). The only publication that exists on L. hoi was by Dippenaar et al. (2001) and described the morpho- logical features of this species by means of scan- ning electron micrographs and drawings. The aim of this study is to contribute to the understanding of the ecology of this parasite on L. polylepis. MATERIALS AND METHODS Sampling sites and hosts Specimens of four Labeobarbus polylepis (Bou- lenger, 1907) populations were collected from In ko- mati and Phongolo River systems within the natural geographical distribution range of the species. Twenty nine specimens were collected from the Phon golo River (27°22’17.93” S, 30°35’24.50” E) and 30 specimens from the Assegaai River (27°4’48.22” S, 30°49’15.09” E) during June 2006. In the Inkomati system, 25 specimens were sam- pled from the Elands River (25°36’56.09” S, 30°30’55.29” E) in March 2005 and 20 in October 2006. Twenty specimens were collected from the Komati River (25°53’40.94” S, 30°17’1.19” E) in January 2007 and 27 in June 2008 (Fig. 1). Collection of fish and parasites Fish specimens were captured by means of gill nets and killed by severing the spinal cord behind the head. They were weighed and their sizes were de- termined in the field by measuring their total length. The fish were dissected by cutting from the anus towards the head and the sex was determined. The gills were removed, dissected apart, placed in water and examined for parasites with the aid of a dissect- ing microscope. The gill arches were divided into three areas: anterior, median and posterior, and the positions of the parasites on each gill arch were noted. Females of L. hoi were collected and fixed in warm alcohol-formaldehyde-acetic acid (AFA) and preserved in 70 % ethanol. FIG. 1 Map indicating the geographic distribution of Labeobarbus polylepis sampled in four rivers (Elands, Phongolo, Assegaai and Komati rivers) in Mpumalanga, South Africa. Dots indicate the position of the sampling sites South Africa South Africa Nooitgedacht Dam Kwena Dam Elands River Ngodwana Dam Ngodwana River Vygeboom Dam Crocodile River Komati River Phongolopoort Dam Swaziland Mozambique Phongolo River Heyshope Dam Assegaai River Usutu River N 229 A. AUSTIN & A. AVENANT-OLDEWAGE Identification of parasites The identification of the parasites took place in the laboratory of the University of Johannesburg. Para- sites were identified as L. hoi based on their mor- phology (Dippenaar et al. 2001). Three develop- mental stages were observed in L. hoi females. Inseminated females were categorized as immature when attaching to the gill arches of the host. When the ovaries became conspicuous the females were considered mature. Subsequently, fertilized eggs are stored in egg sacs hanging from the body and females were classified as gravid. Data presentation Lamproglena hoi specimens were counted and the data obtained were graphically represented. A com- parison between the infections in the four L. poly- lepis populations was done. Geographical locality, seasonality and gender specificity of L. hoi were de- termined. The location of the parasites on the gills was recorded. The correlations between the total length of the host and the number of parasites were determined. Infection levels were expressed as prevalence, mean intensity and abundance accord- ing to the definitions of Bush, Lafferty, Lotz & Shos- tak (1997). RESULTS The combined surface area of the second and third gills of L. polylepis is larger than that of the first and fourth gills (Fig. 2) and the fact that the former two gills are situated centrally in the gill chamber allows for maximum water flow over this area. Lamproglena hoi congregates on the outside of the gill filaments and attaches close to the gill arches with the posterior end facing away from gill arches (Fig. 3A and 3B). In young females the gut content appears green (Fig. 3B) and no proliferation of gill tissue is apparent. In mature females the gut is filled with a red substance that appears to be blood (Fig. 3A) and proliferation of gill tissue is observed (Fig. 3B). Infection statistics of Lamproglena hoi Prevalence, abundance (relative density) and mean intensity Sampling in the Phongolo and Assegaai rivers was done in winter, while the specimens in the Elands River were collected in autumn and spring and in the Komati River in summer and winter. The prev- alence, mean intensity and abundance of the four rivers show seasonal variance (Fig. 7). The prev- alence of L. hoi on the gills of L. polylepis varied between the different rivers. The prevalence in L. FIG. 2 Gill arches of Labeobarbus polylepis in the gill cham- ber. Numbers indicate the gills’ position in succession from anterior to posterior FIG. 3 (A) Photograph of a gill arch of Labeobarbus polylepis to indicate attachment sites of Lamproglena hoi para- sites. Gill is divided into three sites (anterior (A), medi- an (M) and posterior (P)), and (B) an enlargement to show attachment and proliferation at attachment site. Arrows indicate proliferation A B 230 Lamproglena hoi infection on Bushveld Smallscale Yellowfi sh, Labeobarbus polylepis (Boulenger, 1907) polylepis in the Phongolo River (21 %) and Assegaai River (40 %) were relatively low, compared to the considerable higher prevalence in the Elands River (52 % and 95 %) and Komati River (63 % and 75 %). The highest mean intensity value recorded for L. hoi was in the Elands River and increased from 12.92 in autumn to 27.74 in spring (Fig. 4). The lowest val- ues noted were 3.83 in the Assegaai River and 4.17 Elands River Autumn 2005 Elands River Spring 2006 0 10 20 30 40 50 60 70 80 90 100 V a lu e Prevalance (%) Mean intensity (no.) Abundance (no.) FIG. 4 Graph depicting the prevalence, mean intensity and abundance of Lamproglena hoi on the gills of Labeobarbus polylepis in au- tumn 2005 and spring 2006 in the Elands River in Mpumalanga, South Africa V a lu e Prevalance (%) Mean intensity (no.) Abundance (no.) Phongolo River Winter 2006 Assegaai River Winter 2006 0 5 10 15 20 25 30 35 40 45 FIG. 5 Graph depicting the prevalence, mean intensity and abundance of Lamproglena hoi on the gills of Labeobarbus polylepis in win- ter 2006 in the Phongolo and As se gaai rivers in Mpumalanga, South Africa V a lu e Prevalance (%) Mean intensity (no.) Abundance (no.) Komati River Summer 2007 Komati River Winter 2008 0 10 20 30 40 50 65 70 80 FIG. 6 Graph depicting the prevalence, mean intensity and abundance of Lamproglena hoi on the gills of Labeobarbus polylepis in sum- mer 2007 and winter 2008 in the Komati River in Mpu ma langa, South Africa 231 A. AUSTIN & A. AVENANT-OLDEWAGE in the Phongolo River during winter (Fig. 5). The rest of the values ranged between 7.73 (summer) and 10.11 (winter) in the Komati River (Fig. 6). For this fish species, the abundance values of L. hoi ranged from 6.72–26.35 (autumn and spring) in the Elands River to 0.86–1.53 (winter) in the Phongolo and Assegaai rivers and then increased to 5.80– 16.06 (summer and winter) in the Komati River (Fig. 4, 5 and 6). Ecological parameters Seasonality When the data sets from the various surveys are combined a pattern becomes apparent. The highest number of L. hoi was collected in spring. The high- est prevalence, mean intensity and abundance of L. hoi were also recorded in spring. All parameters de- creased toward summer and the lowest numbers were observed during winter surveys (Fig. 7). In autumn 58 % of females were mature; this per- centage increased in winter to 95 % and 96 %, re- spectively with 4 % and 5 % immature, indicating that eggs are not released during winter and that young females are present in this season. In spring 84 % of females were gravid and egg sacs were hanging from the body. In summer 55 % females were gravid, indicating that fertilized eggs start to hatch in spring and con- tinued hatching into the summer season, as gravid females decreased in number from spring to sum- mer (Fig. 8) and non parasitic nauplii were observed in close proximity to the egg sacks. V a lu e Autumn 2005 Summer 2007 0 10 20 30 40 50 65 70 100 90 80 Winter 2006 Winter 2006 Spring 2006 Winter 2008 Seasonal variance Prevalance (%) Mean intensity (no.) Abundance (no.) P e rc e n ta g e ( % ) Mature Immature Gravid 0 10 20 30 40 50 60 70 80 90 100 Elands River Autumn 2005 Elands River Spring 2006 Phongolo River Winter 2006 Assegaai River Winter 2006 Komati River Summer 2007 FIG. 7 Graph illustrating the seasonal variance of prevalence, mean in tensity and abundance of Lamproglena hoi on the gills of Labeobarbus polylepis found in four rivers in Mpumalanga, South Africa FIG. 8 Graph showing composition of immature, mature and gravid Lamproglena hoi females per- centages in autumn, winter, spring and summer 232 Lamproglena hoi infection on Bushveld Smallscale Yellowfi sh, Labeobarbus polylepis (Boulenger, 1907) Location of parasites on the gills The gill arches were divided into three areas: ante- rior, median and posterior and the positions of the parasites on each gill arch were noted. From a sub sample, a total of 729 parasites were collected on the gills of L. polylepis, 49 % were found on the left gills and 51 % on the right gills. On both the left and right side, 79 % of the parasites were attached to the median part of the gill arch. The gill arch’s ante- rior gill area had 14 % of parasites and the posterior gill area 8 %. The second gill on both the left and right sides had the highest numbers of parasites (Fig. 9). The position of L. hoi on the hosts collected from the Elands River in autumn 2005 was exclud- ed from the sub sample. Gender specificity In the sub sample (excluding Eland River data 2005) of 126 infected fish, the number of infected males (n = 56 or 81 % of infected fish) was much higher than that of the females (n = 13 or 19 %). However, mainly male fish were collected in the Komati River, hence the large infection on male hosts (n = 31). Host size preference No correlation existed between the number of para- sites and the size of the host (Fig. 10). DISCUSSION The highest number of L. hoi females was observed during spring and it decreased from summer to au- tumn. The lowest number of parasites occurred in winter. Abundance and mean intensity values differ, the numbers in both cases were lower in spring time (Komati River) than in autumn (Elands River). La- beobarbus polylepis in the Elands River demon- strated extremely high numbers of L. hoi infections during autumn and spring. The hosts in the Komati 0 20 40 60 80 100 120 Location of parasites on gills 11 37 5 19 103 11 13 72 2 6 73 4 10 49 3 15 87 6 20 83 10 7 69 14 A M P A M P A M P A M P A M P A M P A M P A M P L2 L3 L4 R1 R2 R3 R4 N u m b e r o f L a m p ro g le n a h o i 0 10 20 30 40 50 65 70 100 90 80 Length of fish (mm) y = 0.0011x + 13.777 R2 = 2–05 0 100 200 300 400 500 600 N u m b e r o f L a m p ro g le n a h o i FIG. 9 Graph showing the position (an- terior (A), median (M) or poste- rior (P)) of the Lamproglena hoi on the left (L) and right gill (R) arches of Labeobarbus polylepis FIG. 10 Graph showing the number of Lamproglena hoi compared to the total size of Labeobarbus poly lepis hosts 233 A. AUSTIN & A. AVENANT-OLDEWAGE River showed exceptionally high numbers of para- sites in winter compared to those in the Phongolo and Assegaai rivers. Khan & Thulin (1991) suggest- ed that since ectoparasites are directly exposed to the river water, they are in contact with pollutants in it, which could reduce continued occurrence or re- productive tempo. According to Marx & Avenant- Oldewage (1996) and Avenant-Oldewage (2003) higher numbers of ectoparasites occur in less con- taminated waters, while more polluted water has the reverse effect. The Elands River is geographically isolated from the Komati River by two waterfalls, but the sampling localities in both rivers are not highly impacted (Ferreira, Wepener & Van Vuren 2008). The Komati River drainage area is mainly covered by forests and the river is therefore not impacted by anthropogenic activities. The less polluted Elands and Komati riv- ers have higher numbers of copepod ecto par a sites than those fish in the Phongolo and Assegaai rivers. This finding corroborates the results of Avenant- Oldewage (2003) for L. clariae in the Oli fants River and that of Tsotetsi, Avenant-Oldewage & Mashego (2004) on L. clariae in the Vaal River. Only three developmental stages of the life cycle of L. hoi females were observed. The immature stage occurred in winter and the highest percentage of mature females was also observed during this sea- son. In spring, gravid females dominated in num- bers on the gills, and this phenomenon continues into summer. It appears that fertilized eggs hatch in spring and summer. The females die after egg pro- duction as is evident from the decline in the number of gravid females as spring goes into summer (84 % compared to 55 %). There were no significant differences in the parasite load between the left and right gill, which indicates that L. hoi do not have a gill side preference. Rhode (1993) speculated that the preference for a particu- lar gill side is due to the asymmetrical body shape of many parasites. Lam proglena hoi is bilaterally sym- metrical which may possibly explain why there is no gill side preference. Rhode (1993) furthermore sug- gested that the distribution of parasites on different gills differs because of the preference or size of the gills. Marx & Avenant-Oldewage (1996) found that L. clariae concentrated near the ends of the gill arches which are completely different from L. hoi which congregates all over the gill surface. Further- more, Tsotetsi et al. (2004) reported that L. clariae specimens attach midway along the gill filament so that the genital segment is in line with the apex of the filament. In L. hoi, the parasite attaches close to the bony part of the gill arch as well as along the length of the filament towards the tip. Seventy-nine percent of the L. hoi were attached to the median part of the gill arch. This differs from the findings of Tsotetsi et al. (2004), who determined that 52 % L. clariae parasites attach to the median part of gill arch. According to Tsotetsi et al. (2004) the anterior part of the Clarias gariepinus gill arch also harboured 14 % of parasites, similar to the finding in this study. The posterior gill arches of L. poly le pis harboured fewer L. hoi parasites (8 %) compared to L. clariae (34 %) were situated on the pos terior gill arches of C. gariepinus (Tsotetsi et al. 2004). More L. hoi parasites preferred the median position on the gill arches. The second gill arch on both sides had more L. hoi parasites than any other gills. The increased gill surface and water flow through the gill chamber might explain why there are more L. hoi parasites in these positions. It is suggested that wa- ter flow in these areas provides the ideal opportu- nity for attachment. This will furthermore provide an advantage for distribution of offspring. On the other hand, Tsotetsi et al. (2004) found a higher occur- rence of L. clariae on the fourth gill on both sides. The fourth gill arch of C. gariepinus is shorter than the others and consists of fewer, thinner gill fila- ments. Tsotetsi et al. (2004) suggested that the pro- tection or diminished turbulence in the part of the gill chamber may offer an explanation for attachment location. In 2004 Tsotetsi et al. found that L. clariae had no preference regarding the sex of the host. Mainly male fish were collected during this study in the Komati River which makes this sample survey bi- ased and a conclusion could not be made on gen- der preference. Data of the present study showed no correlation be- tween the number of parasites and the host size. This is in agreement with the results of Tsotetsi et al. (2004) on L. clariae on C. gariepinus and those of Sproston, Yin & Hu (1950) on Ophiocephalus ar- gus and number of Lamproglena chinensis. Similar results for other copepod studies have been report- ed by Marcogliese (1991) for Caligus Műller, 1758 and Lo, Morand & Galzin (1998) for Lernaea cypri- nacea Linnaeus, 1758. The increase in the size of the host will lead to an increase in gill surface and volume water flow through the gill chamber. How- ever, these advantages have no affect on the num- ber of parasites. 234 Lamproglena hoi infection on Bushveld Smallscale Yellowfi sh, Labeobarbus polylepis (Boulenger, 1907) CONCLUSION The numbers of L. hoi specimens on the gills of L. polylepis from the four rivers indicate a relatively high prevalence, mean intensity and abundance, and the occurrence of a seasonal variance. These observations corroborate those of Tsotetsi et al. (2004) for L. clariae. The median part of the gill arch on the second gill on both the left and right sides was the preferred attachment site. No correlation was found between the number of parasites and the host size or sex. ACKNOWLEDGEMENTS The authors acknowledge the financial assistance provided by the Water Research Commission and the University of Johannesburg. We express our gratitude to Gordon O’Brien and Quinton Tam for their assistance in the field and laboratory. Gina Walsh is thanked for Fig. 2 and Quinton Tam for Fig. 3. REFERENCES AVENANT-OLDEWAGE, A. 2003. Lamproglena and Lernaea (Copepoda) as possible bio-indicators of environmental de- terioration in the Olifants River. Journal of South African Vet- erinary Association, 72:96. BUSH, A.O., LAFFERTY, K.D., LOTZ, J.M. & SHOSTAK, A.W. 1997. Parasitology meets ecology on its own terms: Margolis et al. revisited. Journal of Parasitology, 83:575–583. DIPPENAAR, S.M., LUUS-POWELL, W.J. & ROUX, F. 2001. Lam proglena hoi n. sp. (Copepoda: Lernaeidae) from two yellowfish hosts, Barbus marequensis and Barbus polylepis, caught in a river in Mpumalanga, South Africa. Onderstepoort Journal of Veterinary Research, 68:209–215. 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Mikrographische Beiträge zur Na- tur geschichte der wirbellosen Tiere. Zweites Heft. Berlin: G. 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