Scanning Electron Microscopy and First Molecular Data of Two Species of Lamproglena (Copepoda: Lernaeidae) from Labeo victorianus (Cyprinidae) and Clarias gariepinus (Clariidae) in Kenya.

A parasitological study carried out in May 2022 and March 2023 in the Nyando River of Lake Victoria Basin, Kenya, disclosed two parasitic lernaeid copepods: Lamproglena cleopatra Humes, 1957, from the gills of a cyprinid, the Ningu Labeo victorianus Boulenger, 1901, endemic to the Lake Victoria drainage system, and Lamproglena clariae Fryer, 1957, from a clariid, the North African catfish Clarias gariepinus (Burchell, 1822). The copepods were studied and supplementary taxonomic information was presented using scanning electron micrographs and genetic data. Scanning electron microscopy (SEM) provided information on the morphology of L. cleopatra's antennae, oral region, thoracic legs (2-5), and furcal rami not previously reported. Analyses of the partial fragments of 18S and 28S rDNA and cytochrome c oxidase subunit 1 (cox1) of the two parasites showed them to be distinct from all other Lamproglena taxa retrieved from GenBank. This study presents new taxonomic information on morphology using SEM and provides the first ribosomal (18S and 28S rDNA) and mitochondrial (mtDNA) data for these two parasite species. The cox1 data provided are the first for all 38 nominal species of Lamproglena. Notably, the study also provides a new host record for L. cleopatra and extends the geographical information of this species to Kenya.

Integrated morphological and molecular characterization of the fish parasitic nematode Rhabdochona (Rhabdochona) gendrei Campana-Rouget, 1961 infecting Labeobarbus altianalis (Boulenger, 1900) in Kenya.

During a parasitological survey carried out between May and August 2022 in the River Nyando, Lake Victoria Basin, a single species of Rhabdochona Railliet, 1916 (Nematoda: Rhabdochonidae) was recorded from the intestine of the Rippon barbel, Labeobarbus altianalis (Boulenger, 1900) (Cyprinidae). Based on light microscopy (LM), scanning electron microscopy (SEM) and DNA analyses the parasite was identified as Rhabdochona (Rhabdochona) gendrei Campana-Rouget, 1961. Light microscopy, SEM and DNA studies on this rhabdochonid resulted in a detailed redescription of the adult male and female. The following additional taxonomic features are described in the male: 14 anterior prostomal teeth; 12 pairs of preanal papillae: 11 subventral and one lateral; six pairs of postanal papillae: five subventral and one lateral, with the latter pair at the level of first subventral pairs when counted from the cloacal aperture. For the female: 14 anterior prostomal teeth and the size and absence of superficial structures on fully mature (larvated) eggs dissected out of the nematode body. Specimens of R. gendrei were genetically distinct in the 28S rRNA and cytochrome c oxidase subunit 1 (cox1) mitochondrial gene regions from known species of Rhabdochona. This is the first study that provides genetic data for a species of Rhabdochona from Africa, the first SEM of R. gendrei, and the first report of this parasite from Kenya. The molecular and SEM data reported herein provide a useful point of reference for future studies on Rhadochona in Africa.

A histological evaluation of pansteatitis-affected Mozambique tilapia, Oreochromis mossambicus (Peters 1852), from different geographical locations in South Africa.

Pansteatitis is the leading cause for the decline in Nile crocodile populations and the sporadic mortality of fish in the Olifants River System, South Africa. To determine the prevalence of this disease in lentic systems, Mozambique tilapia, Oreochromis mossambicus, were collected from Lake Loskop, Lake Flag Boshielo, Phalaborwa Barrage and Lake Luphephe-Nwanedi. The former three impoundments are located within the main stem of the Olifants River, while the latter, which is geographically isolated and situated in the Limpopo River System, served as a reference site. Mesenteric adipose, liver, serosa of the swim bladder, gill and the skeletal muscle of fish sampled were examined for gross and microscopic evidence of pansteatitis. Microscopically observed changes were used to statistically compare pansteatitis prevalence between samples and sites. Based on histopathological evaluation, the adipose tissue in the liver, swim bladder serosa and coelom from severely debilitated individuals showed the most significant pathological changes. Lesions indicative of steatitis were observed in fish collected from Lake Loskop (75%), Lake Flag Boshielo (22%) and Lake Luphephe-Nwanedi (15%). Further investigation is warranted to understand the pervasiveness and mechanisms driving pathological changes of pansteatitis at Lake Flag Boshielo, Phalaborwa Barrage and Lake Luphephe-Nwanedi.

Occurrence and characterisation of tongue worms, Linguatula spp., in South Africa.

A total of 509 mammalian vertebrates, belonging to 76 species, were examined for infection with pentastomid parasites. These animals were from 8 of the 9 provinces in South Africa. Linguatulid pentastomes were found only in 7 animals, specifically the African Lion (n = 3) and African Buffalo (n = 4). Adult parasites were found in the lion but nymphs, of various stages, were found in the buffalo. A detailed morphological examination of adult parasites using both light and scanning electron microscopy techniques suggested the specimens were Linguatula nuttalli Sambon1922. Sequences of 18S ribosomal DNA and Cox1 regions obtained from both adult and nymph stages suggested they belong to the one species. Phylogenetic analyses of Linguatula spp. based on the 18S and Cox1 sequences available in GenBank and obtained in the present study showed a clear distinction between L. nuttalli, L. arctica and L. serrata (from Europe and Australia). Several specimens from the Palearctic region which were previously assumed to be L. serrata formed a distinct group in the phylogenetic tree suggesting they probably belong to a different, and as of yet, unknown species.

Lipidomics for wildlife disease etiology and biomarker discovery: a case study of pansteatitis outbreak in South Africa.

INTRODUCTION: Lipidomics is an emerging field with great promise for biomarker and mechanistic studies due to lipids diverse biological roles. Clinical research applying lipidomics is drastically increasing, with research methods and tools developed for clinical applications equally promising for wildlife studies. OBJECTIVES: Limited research to date has applied lipidomics, especially of the intact lipidome, to wildlife studies. Therefore, we examine the application of lipidomics for in situ studies on Mozambique tilapia (Oreochromis mossambicus) in Loskop Dam, South Africa. Wide-scale mortality events of aquatic life associated with an environmentally-derived inflammatory disease, pansteatitis, have occurred in this area. METHODS: The lipidome of adipose tissue (n = 31) and plasma (n = 51) from tilapia collected from Loskop Dam were characterized using state of the art liquid chromatography coupled to high-resolution tandem mass spectrometry. RESULTS: Lipid profiles reflected pansteatitis severity and were significantly different between diseased and healthy individuals. Over 13 classes of lipids associated with inflammation, cell death, and/or oxidative damage were upregulated in pansteatitis-affected adipose tissue, including ether-lipids, short-chained triglyceride oxidation products, sphingolipids, and acylcarnitines. Ceramides showed a 1000-fold increase in the most affected adipose tissues and were sensitive to disease severity. In plasma, triglycerides were found to be downregulated in pansteatitis-affected tilapia. CONCLUSION: Intact lipidomics provided useful mechanistic data and possible biomarkers of pansteatitis. Lipids pointed to upregulated inflammatory pathways, and ceramides serve as promising biomarker candidates for pansteatitis. As comprehensive coverage of the lipidome aids in the elucidation of possible disease mechanisms, application of lipidomics could be applied to the understanding of other environmentally-derived inflammatory conditions, such as those caused by obesogens.

Morphological Studies of Developmental Stages of Oculotrema hippopotami (Monogenea: Polystomatidae) Infecting the Eye of Hippopotamus amphibius (Mammalia: Hippopotamidae) Using SEM and EDXA with Notes on Histopathology.

The present study was performed to observe histopathological effects of Oculotrema hippopotami Stunkard, 1924 infection in the eye of Hippopotamus amphibius, as well as to reveal new details of morphology and structural features of this monogenean and its comparison between 2 age stages of the parasite. This was done using both light and scanning electron microscopy, energy dispersive X-ray analysis (EDXA) and histopathology. The presence of a mixture of different generations (adult and sub-adult) in one host individual is common for Oculotrema Stunkard, 1924 in contrast to Polystoma Zeder, 1800. New metrical and graphical information obtained for adults and sub-adults compared with the previous studies. Here we show the presence of genital papillae in adults, metrical data on the distal part of the vas deferens. SEM micrographs of sperm ejaculatory structures and information about the flattened dorsal side of the body provided for the first time. Histopathological changes, such as necrosis and hemorrhage in host tissues as a result of O. hippopotami attachment structures are described. Structural analysis of different body parts of O. hippopotami of both age groups are also included. We show qualitative differences in the presence of hardening ions (S, P, Ca) in attachment structures (oral and haptor suckers) that increase with the age of the worm. The presence of sub-adults and adults on the same host, together with high levels of infection without high pathogenicity may account for Oculotrema being one of the most successful parasites among the Monogenea.

Tissue distribution of perfluoroalkyl acids and health status in wild Mozambique tilapia (Oreochromis mossambicus) from Loskop Dam, Mpumalanga, South Africa.

This study examined concentrations of 15 perfluoroalkyl acids (PFAAs) in tissues from male Mozambique tilapia (Oreochromis mossambicus) collected at Loskop Dam, Mpumalanga, South Africa in 2014 and 2016. Nine of the 15 PFAAs were detected frequently and were included in statistical analysis and included two of the most commonly known PFAAs, perfluorooctanesulfonic acid (PFOS) (median, 41.6ng/g) and perfluorooctanoic acid (PFOA) (median, 0.0825ng/g). Of the tissues measured, plasma (2016 and 2014 median, 22.2ng/g) contained the highest PFAA burden followed by (in descending order): liver (median, 11.6ng/g), kidney (median, 9.04ng/g), spleen (median, 5.92ng/g), adipose (median, 2.54ng/g), and muscle (median, 1.11ng/g). Loskop Dam tilapia have been affected by an inflammatory disease of the adipose tissue known as pansteatitis, so this study also aimed to investigate relationships between PFAA tissue concentrations and incidence of pansteatitis or fish health status. Results revealed that healthy tilapia exhibited an overall higher (p-value<0.05) PFAA burden than pansteatitis-affected tilapia across all tissues. Further analysis showed that organs previously noted in the literature to contain the highest PFAA concentrations, such as kidney, liver, and plasma, were the organs driving the difference in PFAA burden between the two tilapia groups. Care must be taken in the interpretations we draw from not only the results of our study, but also other PFAA measurements made on populations (human and wildlife alike) under differing health status.

Guppies (Poecilia reticulata) introducing an alien parasite, Camallanus cotti (Nematoda: Camallanidae) to Africa, the first report.

Introduced alien fish species and their associated parasites may result in a serious threat to indigenous biodiversity. Furthermore, this may have negative impacts on cultured fish as well as on native parasitic fauna. In the present study, the invasive Asian nematode, Camallanus cotti Fujita, 1927 (Nematoda: Camallanidae), is reported from the guppy (Poecilia reticulata) for the first time in Africa. This parasite is assumed to be introduced into Africa along with the introduction of exotic poeciliid fishes, which are known to be the most common hosts of C. cotti in ornamental fish industry worldwide.The presence of this parasite in both aquarium-cultured fish as well as fish from natural waterbodies is evidence of the introduction of the alien organisms due to insufficient prophylactic veterinary control during transfer of non-native hosts between countries and the spread of them by the anthropogenic introduction to natural systems.

Parasites of domestic and wild animals in South Africa. L. Ixodid ticks infesting horses and donkeys.

The aim of the study was to determine the species spectrum of ixodid ticks that infest horses and donkeys in South Africa and to identify those species that act as vectors of disease to domestic livestock. Ticks were collected opportunistically from 391 horses countrywide by their owners or grooms, or by veterinary students and staff at the Faculty of Veterinary Science, University of Pretoria. Ticks were also collected from 76 donkeys in Limpopo Province, 2 in Gauteng Province and 1 in North West province. All the ticks were identified by means of a stereoscopic microscope. Horses were infested with 17 tick species, 72.1% with Rhipicephalus evertsi evertsi, 19.4% with Amblyomma hebraeum and 15.6% with Rhipicephalus decoloratus. Rhipicephalus evertsi evertsi was recovered from horses in all nine provinces of South Africa and R. decoloratus in eight provinces. Donkeys were infested with eight tick species, and 81.6% were infested with R. evertsi evertsi, 23.7% with A. hebraeum and 10.5% with R. decoloratus. Several tick species collected from the horses and donkeys are the vectors of economically important diseases of livestock. Rhipicephalus evertsi evertsi is the vector of Theileria equi, the causative organism of equine piroplasmosis. It also transmits Anaplasma marginale, the causative organism of anaplasmosis in cattle. Amblyomma hebraeum is the vector of Ehrlichia ruminantium, the causative organism of heartwater in cattle, sheep and goats, whereas R. decoloratus transmits Babesia bigemina, the causative organism of babesiosis in cattle.

First Introduction of Two Australian Temnocephalan Species into Africa with an Alien Host: Double Trouble.

The redclaw crayfish, Cherax quadricarinatus (Decapoda: Parastacidae), is native to Australia but has been introduced to South Africa as a warm-water aquaculture species. In a South African natural waterbody, examined crayfish had high-intensity infections of 3 temnocephalan species on their body surfaces and within the branchial chambers. Temnocephalans were characterized using light and scanning electron microscopy and identified as Craspedella pedum, Diceratocephala boschmai, and Didymorchis sp. This is the first report of the introduction of Australian temnocephalans, C. pedum and Didymorchis sp., to Africa and expands the known distribution of these species beyond their presumptive native range. The present study also documents a naturalized population of C. quadricarinatus from a natural water body in South Africa, comprising a new geographical locality record.

Perfluorinated alkyl acids in the plasma of South African crocodiles (Crocodylus niloticus).

Perfluorinated alkyl acids (PFAAs) are environmental contaminants that have been used in many products for over 50 years. Interest and concern has grown since 2000 on the widespread presence of PFAAs, when it was discovered that PFAAs were present in wildlife samples around the northern hemisphere. Since then, several studies have reported PFAAs in wildlife from many locations, including the remote regions of Antarctica and the Arctic. Although there are a multitude of studies, few have reported PFAA concentrations in reptiles and wildlife in the Southern Hemisphere. This study investigated the presence of PFAAs in the plasma of Nile crocodiles (Crocodylus niloticus) from South Africa. Crocodiles were captured from five sites in and around the Kruger National Park, South Africa, and plasma samples examined for PFAAs. Perfluorooctane sulfonate (PFOS) was the most frequent PFAA detected; with median values of 13.5 ng/g wet mass in crocodiles. In addition to PFOS, long chain perfluorinated carboxylic acids were also detected. Correlations between total length and PFAA load were investigated, as were differences in PFAA accumulation between sexes. No correlations were seen between crocodile size, nor were there sex-related differences. Spatial differences were examined and significant differences were observed in samples collected from the different sites (p < 0.05). Flag Boshielo Dam had the highest PFOS measurements, with a median concentration of 50.3 ng/g wet mass, when compared to the other sites (median concentrations at other sites below 14.0 ng/g wet mass). This suggests a point source of PFOS in this area.

Predictive Blood Chemistry Parameters for Pansteatitis-Affected Mozambique Tilapia (Oreochromis mossambicus).

One of the largest river systems in South Africa, the Olifants River, has experienced significant changes in water quality due to anthropogenic activities. Since 2005, there have been various "outbreaks" of the inflammatory disease pansteatitis in several vertebrate species. Large-scale pansteatitis-related mortality events have decimated the crocodile population at Lake Loskop and decreased the population at Kruger National Park. Most pansteatitis-related diagnoses within the region are conducted post-mortem by either gross pathology or histology. The application of a non-lethal approach to assess the prevalence and pervasiveness of pansteatitis in the Olifants River region would be of great importance for the development of a management plan for this disease. In this study, several plasma-based biomarkers accurately classified pansteatitis in Mozambique tilapia (Oreochromis mossambicus) collected from Lake Loskop using a commercially available benchtop blood chemistry analyzer combined with data interpretation via artificial neural network analysis. According to the model, four blood chemistry parameters (calcium, sodium, total protein and albumin), in combination with total length, diagnose pansteatitis to a predictive accuracy of 92 percent. In addition, several morphometric traits (total length, age, weight) were also associated with pansteatitis. On-going research will focus on further evaluating the use of blood chemistry to classify pansteatitis across different species, trophic levels, and within different sites along the Olifants River.