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Bartonella spp. are intracellular bacteria associated with several re-emerging human diseases. Small mammals play a significant role in the maintenance and spread of Bartonella spp. Despite the high small mammal biodiversity in South Africa, there is limited epidemiological information regarding Bartonella spp. in these mammals. The main aim of this study was to determine the prevalence and genetic diversity of Bartonella spp. from wild small mammals from 15 localities in 8 provinces of South Africa. Small mammals ( n = 183) were trapped in the Eastern Cape, Free State, Gauteng, Limpopo, Mpumalanga, Northern Cape, North West, and Western Cape provinces of South Africa between 2010 and 2018. Heart, kidney, liver, lung, and spleen were harvested for Bartonella DNA screening, and prevalence was determined based on the PCR amplification of partial fragments of the 16S-23S rRNA intergenic spacer (ITS) region, gltA, and rpoB genes. Bartonella DNA was detected in Aethomys chrysophilus , Aethomys ineptus , Gerbillurus spp., Lemniscomys rosalia , Mastomys coucha , Micaelamys namaquensis , Rhabdomys pumilio, and Thallomys paedulcus . An overall prevalence of 16.9% (31/183, 95% CI: 12.2%-23%) was observed. Bartonella elizabethae , Bartonella grahamii, and Bartonella tribocorum were the zoonotic species identified, while the remaining sequences were aligned to uncultured Bartonella spp. with unknown zoonotic potential. Phylogenetic analyses confirmed five distinct Bartonella lineages (I-V), with lineage IV displaying strong M. coucha host specificity. Our results confirm that South African wild small mammals are natural reservoirs of a diverse assemblage of Bartonella spp., including some zoonotic species with high genetic diversity, although prevalence was relatively low.IMPORTANCESmall mammals play a significant role in the maintenance and spread of zoonotic pathogens such as Bartonella spp. Despite the high small mammal biodiversity in southern Africa including South Africa, there is limited epidemiological information regarding Bartonella spp. in these mammals across the country. Results from our study showed the liver and spleen had the highest positive cases for Bartonella spp. DNA among the tested organs. Bartonella elizabethae , B. grahamii, and B. tribocorum were the three zoonotic species identified and five distinct Bartonella lineages (I-V) were confirmed through phylogenetic analyses. To the best of our knowledge, this study presents the first extensive nuclear diversity investigation of Bartonella spp. in South African small mammals in South Africa., Competing Interests: The authors declare no conflict of interest.

The systematic review and meta-analysis were conducted to determine the estimates of the prevalence and infection rates of natural and experimental infections of amphistome species in intermediate host snails (IHs) across different continents. A search of peer-reviewed literature on natural and experimental infections of freshwater snails with amphistome species was conducted from four electronic databases from 1984 to 2023. The estimates of the prevalence and/or infection rates were based on 36 eligible peer-reviewed articles, which met the inclusion criteria and reported on natural and experimental infections of amphistome species in freshwater snails. The results showed that a total of 1,67,081 snail species from the peer-reviewed articles were examined for natural infections and 7,659 snail species for experimental infections. The overall pooled prevalence of amphistome infections from naturally infected snails was 2% (95% CI: 0-4), while the overall pooled prevalence of amphistome infections from infections was 40% (95% CI: 18-64). The highest pooled prevalence of natural infection was 3%, which was recorded in Europe (95% CI: 1-7%). The highest overall prevalence of naturally infected amphistome was 6% (95% CI: 0-20%) for Paramphistomum epiclitum . The Americas had the highest pooled prevalence of experimental amphistome infection among freshwater snails (66%; 95% CI: 26-96%). The highest pooled infection rate of 65% (95% CI: 12-100%) was recorded for Paramphistomum cervi in experimental infections. Galba truncatula was the only snail that qualified for meta-analysis for natural infection with Calicophoron daubneyi, with a pooled prevalence of 3% (95% CI: 1-8%). Galba truncatula infected with C. daubneyi and P. cervi, and Bulinus tropicus infected with Calicophoron microbothrium in the experimental infection qualified for the meta-analysis, with an overall infection rate of 66% (95% CI: 34-92%) and 30% (95% CI: 0-74%), respectively. The pooled prevalence of amphistome species infection in the intermediate host (IH) snails based on detection techniques was higher with PCR compared to the dissection and shedding of cercariae. The results from the quality effects model revealed a high heterogeneity and publication bias between studies. This meta-analysis provided valuable insights into the prevalence and infection rates of amphistome species in snail IHs across different geographical regions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Nyagura, Malatji and Mukaratirwa.)

This review summarises studies on distribution, diversity, and prevalence of gastrointestinal helminth infections in wild ruminants in sub-Saharan Africa. The results showed that 109 gastrointestinal tract (GIT) helminth species or species complexes were recorded in 10 sub-Saharan African countries. South Africa reported the highest number of species because most studies were carried out in this country. Eighty-eight nematode species or species complexes were recorded from 30 wild ruminant species across eight countries. The genus Trichostrongylus recorded the highest number of species and utilised the highest number of wild ruminant species, and along with Haemonchus spp., was the most widely distributed geographically. Fifteen trematode species or species complexes were reported from seven countries. The genus Paramphistomum recorded the highest number of species, and Calicophoron calicophoron was the most commonly occurring species in sub-Saharan African countries and infected the highest number of hosts. Six cestode species or species complexes from one family were documented from 14 wild hosts in seven countries. Moniezia spp. were the most commonly distributed in terms of host range and geographically. Impala were infected by the highest number of nematodes, whilst Nyala were infected by the highest number of trematode species. Greater kudu and Impala harbored the largest number of cestodes. The prevalence amongst the three GIT helminths taxa ranged between 1.4% and 100% for nematodes, 0.8% and 100% for trematodes, and 1.4% and 50% for cestodes. There is still limited information on the distribution and diversity of GIT helminths in wild ruminants in most sub-Saharan African countries.

This study aimed at determining the identity of freshwater snails collected from selected water habitats frequented by wildlife as source of drinking water in the Matebeleland region of Zimbabwe and further screening the identified snails for natural infections with amphistomes using PCR. A total of 487 freshwater snails were collected from six areas in the Matebeleland region of Zimbabwe for identification and screening of amphistome infection. Eight freshwater snail species were morphologically identified and Biomphalaria pfeifferi, Bul. tropicus, Bul. truncatus, Bul. globosus , and L. (R.) natalensis were confirmed using the COI gene. Bulinus tropicus and Phy. acuta were the most abundant species at 33.9% (165/487) and 31.2% (155/487), respectively. DNA of amphistome was detected in 11.9% (58/487) of the collected snails. The highest infection rate was detected in Bul. globosus (44.4%). West Nicholson recorded the highest infection rate (33.9%), and infection was not detected in L. (R.) natalensis , Phy. acuta , and Bellamya spp. Amphistome DNA from M. tuberculata was successfully sequenced and identified as Calicophoron microbothrium . An additional band was detected in M. tuberculata , Bul. tropicus , and Bul. trancatus , which showed a 96.42% similarity to Paragonimus sp. sequence in the GenBank.

This review summarizes published records on the prevalence, species diversity, geographical distribution, mixed infections, co-infections with other trematodes and intermediate hosts (IHs) of amphistomes (rumen flukes) of wild ruminants in Africa. Literature search was conducted on Google Scholar, PubMed and JSTOR, using a combination of predetermined search terms and Boolean operators. Of the 54 African countries searched, results showed that occurrence of amphistome infections in wild ruminants have only been reported in 23 countries. A total of 38 amphistome species consisting of the following 11 genera were recorded, viz Bilatorchis , Calicophoron, Carmyerius , Choerecotyloides , Cotylophoron , Explanatum , Gastrothylax , Gigantocotyle, Leiperocotyle , Paramphistomum and Stephanopharynx. These were recorded in 39 wild ruminant species, belonging to the Bovidae family. The genus Carmyerius recorded the highest number of species (n = 13) across nine countries Africa. However, Calicophoron species (n = 9) were more widely distributed, occurring in 17 countries across all regions of Africa. Species of this genus collectively infected 27 wild ruminant species. However, at a species level, Cotylophoron cotylophorum infected the highest number of wild ruminant species. Prevalence of infection based on post-mortem examination ranged from 1.89% in African Buffalo to 100% in Defassa waterbuck from Egypt and Zambia, respectively. The most common mixed infections recorded were those between amphistomes of the same or different genus. Snail intermediate hosts (IHs) were described for 10/38 amphistome species, and these were predominantly species from Plarnobidae family. Despite the richness in diversity of amphistomes infecting wild ruminants in Africa, there is need to further confirm identity of snail IHs and the amphistome species using both morphological and molecular techniques. Furthermore, more studies are recommended to assess the burden of amphistomosis in commercially reared wildlife/game farming, mixed game and livestock farming systems in Africa., Competing Interests: The authors declare no conflict of interest., (© 2024 The Authors.)

This review collated existing data on the occurrence, distribution, and prevalence of haemoparasites of poultry in sub-Saharan Africa. A literature search was conducted on three electronic search databases using search terms and Boolean operators (AND, OR). The results recorded 16 haemoparasites, viz., Leucocytozoon spp., L. marchouxi, L. neavei, L. sabrazesi, L. schoutedeni, Haemoproteus columbae, H. pratasi, Haemoproteus spp., Plasmodium spp., P. gallinaceum, P. circumflexum, P. juxtanucleare, Trypanosoma avium, T. gallinarum, T. numidae, and Hepatozoon spp. from a wide range of poultry species distributed across Nigeria, Kenya, South Africa, Tanzania, Uganda, Botswana, Zimbabwe, Ghana, Cameroon, and Zambia. Infections due to Haemoproteus and Leucocytozoon species were the most common and documented in eight of the ten reviewed countries. The presence of mixed infections was observed in quails, pigeons, chickens, ducks, turkeys, and guineafowls, but predominantly in chickens. Co-infections by Plasmodium spp. and Haemoproteus spp. were the most common, which may be attributed to the distribution of these species, coupled with the availability of vectors they are associated with in areas from which they were documented. The information generated in this review is essential for improving existing preventive and control measures of these parasites in sub-Saharan Africa.

Besnoitiosis is a parasitic disease of economic importance caused by cyst-forming protozoa from the genus Besnoitia . The disease affects the skin, subcutis, blood vessels, and mucous membranes of the animals. It is traditionally endemic in the tropical and sub-tropical regions of the world, and causes enormous economic loss associated with impaired productivity and reproduction, as well as skin lesions. Therefore, knowledge of the epidemiology of the disease, including the current Besnoitia species occurring in sub-Saharan Africa, the wide range of mammalian species hosts they use as intermediate hosts, and the clinical signs manifested by infected animals is crucial in developing effective prevention and control measures. This review collected information from peer-reviewed publications involving the epidemiology and clinical signs of besnoitiosis in sub-Saharan Africa using four electronic databases. Results showed that B. besnoiti , B. bennetti, B. caprae, B. darlingi-like and unidentified Besnoitia spp. were found naturally infecting livestock and wildlife across nine reviewed sub-Saharan African countries. Besnoitia besnoiti was the most common species, occurring in all nine reviewed countries, and utilised a wide range of mammalian species as intermediate hosts. Prevalence of B. besnoiti ranged from 2.0 to 80.3%, and B. caprae 5.45-46.53%. Infection rate was high with serology compared to other techniques. Some of the typical signs of besnoitiosis included sand-like cysts on the sclera conjunctiva, nodules in the skin, thickening and wrinkling of the skin and alopecia. Inflammation, thickening and wrinkling of the scrotum were observed in bulls, and lesions on the scrotum deteriorated progressively and became generalized in some cases in spite of treatment. There is still a need for surveys focusing on detecting and identifying Besnoitia spp. using molecular techniques in combination with serological, histology and visual observation, and scoping their natural intermediate and definitive hosts, as well as assessing the burden of the disease animals reared on different husbandry systems in sub-Saharan Africa., Competing Interests: Authors declares no conflict of interest., (© 2023 The Authors.)

The occurrence of Fasciola gigantica and F. hepatica in Africa is well documented; however, unlike in Asia, there is a paucity of information on the existence of hybrids or parthenogenetic species on the continent. Nonetheless, these hybrid species may have beneficial characteristics, such as increased host range and pathogenicity. This study provides evidence of the potential existence of Fasciola hybrids in Africa. A literature search of articles published between 1980 and 2022 was conducted in PubMed, Google Scholar, and Science Direct using a combination of search terms and Boolean operators. Fasciola species were documented in 26 African countries with F. hepatica being restricted to 12 countries, whilst F. gigantica occurred in 24 countries, identified based on morphological features of adult Fasciola specimens or eggs and molecular techniques. The co-occurrence of both species was reported in 11 countries. However, the occurrence of potential Fasciola hybrids was only confirmed in Egypt and Chad but is suspected in South Africa and Zimbabwe. These were identified based on liver fluke morphometrics, assessment of the sperms in the seminal vesicle, and molecular techniques. The occurrence of intermediate host snails Galba truncatula and Radix natalensis was reported in Ethiopia, Egypt, South Africa, Tanzania, and Uganda, where F. hepatica and F. gigantica co-occurrences were reported. The invasive Pseudosuccinea columella snails naturally infected with F. gigantica were documented in South Africa and Egypt. In Zimbabwe, P. columella was infected with a presumed parthenogenetic Fasciola. This suggests that the invasive species might also be contributing to the overlapping distributions of the two Fasciola species since it can transmit both species. Notwithstanding the limited studies in Africa, the potential existence of Fasciola hybrids in Africa is real and might mimic scenarios in Asia, where parthenogenetic Fasciola exist in most Asian countries. In South Africa, aspermic F. hepatica and Fasciola sp. have been reported already, and Fasciola hybrids have been reported? in Chad and Egypt. Thus, the authors recommend future surveys using molecular markers recommended to identify Fasciola spp. and their snail intermediate hosts to demarcate areas of overlapping distribution where Fasciola hybrids and/or parthenogenetic Fasciola may occur. Further studies should also be conducted to determine the presence and role of P. columella in the transmission of Fasciola spp. in these geographical overlaps to help prevent parasite spillbacks.

This review gathered information from peer-reviewed publications on the epidemiology, prevention and control of gastrointestinal helminths (GIHs) parasites of small ruminants in the Caribbean region from 1990 to 2021. Literature search was performed on four electronic databases using a combination of search terms and Boolean operators. Results showed that gastrointestinal nematodes (GINs) were the most common parasites, with seven genera documented across six reviewed countries. Haemonchus contortus was the most common species occurring in all six countries, and predominant in occasions where mixed infections were observed. Moniezia species were the only Platyhelminth species documented in Trinidad and Grenada. The overall prevalence of GIH infections was observed to be high in goats than in sheep. A high level of anthelminthic resistance (AR) with H. contortus was reported, whilst other nematodes showed to be effectively controlled by one or more anthelmintic drugs. FAMACHA © method was applied in the identification of anaemic animals for selective treatment, hence contributing to the decreased use of anthelmintic drugs. There is a need for national surveys of GIHs and AR in small ruminants in the Caribbean countries. Surveys, coupled with the use of molecular techniques to detect and identify species of GIHs present in the Caribbean region, as well as their epidemiology which will inform development of integrated control strategies is recommended. There is also a need to create awareness to small ruminant farmers in the region on the prevailing challenge of AR and limit wanton use of anthelminthics to reduce or prevent AR in small ruminants., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Free-range chickens are predisposed to diverse parasitic infections during scavenging. Accurate identification of these parasites using morphological characters has been a challenge. Therefore, this study aimed to identify nematodes from the Heterakidae and Ascarididae family infecting free-ranging chickens from KwaZulu-Natal province of South Africa using a combination of morphological and molecular techniques. Forty-two free-ranging adult indigenous chickens were purchased from randomly selected households in Shongweni (n=12), Umzinto (n=10), Gingindlovu (n=10) and Ozwathini (n=10) rural villages and examined for nematodes of the Heterakidae and Ascarididae family. Collected specimen were identified morphologically and confirmed using mitochondrial and nuclear ribosomal markers. Results showed that Ascaridia galli was common, occurring at all sampling locations with an overall prevalence of 58.3%, while Heterakis gallinarum and H. beramporia occurred in three locations. Ascaridia galli had high prevalence in Shongweni (58.3%), followed by Gingindlovu (40%), Ozwathini (20%) and Umzinto (10%). Heterakis gallinarum infection was prevalent in three locations, with an overall prevalence of 90% in Gingindlovu, 80% in Ozwathini and 58.3 % in Shongweni. Heterakis gallinarum and H. beramporia were not recorded in Umzinto. Heterakis beramporia was recorded in low prevalence in Gingindlovu (20%), Ozwathini (10%) and Shongweni (8.3%) villages. Mixed infections of A. galli and H. gallinarum were recorded in Gingindlovu, Ozwathini and Shongweni, and H. gallinarum and H. beramporia in Gingindlovu. Molecular analysis confirmed identification of A. galli, and further showed close relationship with the GenBank-derived South African isolates. Haplotype network further confirmed their ancestral history, where all South African A. galli isolates formed five novel haplotypes corresponding with the structure of the phylogenetic tree. Similar structure was observed with Heterakis isolates, where analysis of the cox1 gene showed that H. gallinarum formed a well-supported monophyletic clade with other Heterakis species. The ITS marker identified three specimens from Gingindlovu, Ozwathini and Shongweni as H. beramporia, which formed strongly supported sister clade to H. indica and this is the first report confirming the occurrence of H. beramporia in South Africa., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

This review was conducted to provide an update on the status of the occurrence of Fasciola species in livestock, wildlife and humans, and the geographical distribution of snail intermediate host (IH) species in South Africa. The literature search was conducted on four electronic databases using the Boolean operators in combination with predetermined search terms for thematic analysis. Results showed that Fasciola species have been reported in six out of nine provinces of South Africa in the last six decades (1960-2021), with both F. hepatica and F. gigantica infecting vertebrate hosts and F. hepatica and Fasciola spp infecting humans. Results also showed that most studies relied on morphological identification of eggs and flukes without molecular confirmation, which might have led to the misidentification of specimens, especially when immature. Fasciola hepatica has been documented in Limpopo, Mpumalanga, and KwaZulu-Natal provinces. The occurrences of Galba truncatula as the probable snail IH for F. hepatica in the three provinces has been documented while Pseudosuccinea columella has only been documented in Mpumalanga and KwaZulu-Natal provinces. The occurrence of F. gigantica to date has been reported in Mpumalanga and KwaZulu-Natal provinces, with overlapping distribution with F. hepatica . Radix natalensis , the main IH of F. gigantica has been documented in all the three provinces, while the two alien Radix species ( R. auricularia and R. rubiginosa ) were documented in KwaZulu-Natal province and have been implicated elsewhere with the transmission of F. gigantica . The presence of Fasciola spp eggs and antibodies in humans were documented in the Eastern Cape and the Western Cape provinces, where both P. columella and G. truncatula are known to be present. The prevalence of Fasciola spp infection in livestock ranged from 9.1 to 37.67 %, with an estimated annual financial loss ranging from R44930.26-129901 in cattle production in the Eastern Cape province of South Africa. This review reaffirms the scarcity of information on the occurrence and burden of fasciolosis in South Africa, and further highlights the importance of future research covering all provinces of the country and assessing the public health significance of the disease in resource-poor livestock communities in the areas where the parasite is endemic., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Nyagura, Malatji and Mukaratirwa.)

Hydatid disease is a parasitic zoonosis caused by genotypes of the genus Echinococcus . This disease inflicts economic loses in livestock and cause public health burden in resource poor mostly in developing countries. The aim of this study was to determine the prevalence and identity of the genotypes responsible for hydatid cysts in cattle, goats and pigs slaughtered at selected abattoirs of southern provinces of Mozambique. Cysts were collected from liver and lungs and hydatid confirmation was made by cystic membrane observation and visualization of protoscoleces by light microscope. Thirty-two hydatid cysts from 817 cattle and two from 68 pigs were collected from local slaughterhouses and slabs. DNA was extracted from protoscoleces of each cyst together with the cystic membrane and amplified based on the mitochondrial subunit 1 of the cox1 and nad1 gene. The overall prevalence of hydatid cysts was 3.9% in cattle, 2.9% in pigs and none of the goats were found with cysts. All cysts collected from cattle and pigs were identified as Echinococcus ortleppi (G5) with a minimum homology of 99% on BLAST analysis. Our results confirm the presence of E. ortleppi in cattle and pigs in southern Mozambique at a low prevalence and further studies are recommended to determine the risk factors favoring the transmission of this zoonotic parasite in the resource-poor livestock farming communities of this region., Competing Interests: Conflict of interestThe authors declare that they have no competing interest., (© Indian Society for Parasitology 2021.)

Pseudosuccinea columella is considered invasive and has become an important intermediate host of both Fasciola species in many regions of the world. This systematic review assessed the geographical distribution of P. columella, and its implications in the transmission of Fasciola hepatica and Fasciola gigantica, globally. A literature search was conducted on Google Scholar, JSTOR and PubMed databases using Boolean operators in combination with predetermined search terms for thematic analysis. Results show that P. columella has been documented in 22 countries from Europe (3), Africa (8), Oceania (2), North America (3) and South America (6). Furthermore, this snail species has shown to adapt to and inhabit a vast array of freshwater bodies including thermal lakes and ditches with acidic soils. Studies showed that P. columella transmits F. hepatica, with natural and experimental infections documented in sub-Saharan Africa, Europe, South America and North America. Experimental infection studies in Cuba showed the presence of P. columella populations resistant to F. hepatica infection. Furthermore, some populations of this invasive snail collected from F. hepatica endemic locations in Brazil, Venezuela, Australia, South Africa, Colombia and Argentina were found without Fasciola infection. As a result, the role played by this snail in the transmission of Fasciola spp. in these endemic areas is still uncertain. Therefore, further studies to detect natural infections are needed in regions/countries where the snail is deemed invasive to better understand the veterinary and public health importance of this snail species in Fasciola-endemic areas and determine the global dispersion of resistant populations of P. columella.

The current study aimed at molecular identification and comparing the diversity of arthropods communities between pig and sheep carcasses during the cold and warm season in KwaZulu-Natal province of South Africa. Adult arthropods found on and around the carcasses were collected using either fly traps or forceps. Molecular analyses confirmed the identification of twelve arthropod species collected from both sheep and pig carcasses during the cold season. Results showed that 11 of 12 arthropod species were common in both sheep and pig carcasses, with exception to Onthophagus vacca (Coleoptera: Scarabaeidae) (Linnaeus, 1767) and Atherigona soccata (Diptera: Muscidae) (Rondani, 1871) species which were unique to sheep and pig carcasses respectively. However, during the warm season, the sheep carcass attracted more arthropod ( n = 13) species as compared to the pig carcass. The difference in the obtained arthropod was due to the presence of O. vacca which was also unique to the sheep carcass during this season. Furthermore, there was an addition of a beetle species Hycleus lunatus (Coleoptera: Meloidae) (Pallas, 1782), which was collected from both sheep and pig carcasses but unique to the warm season. The pig carcass attracted more dipteran flies during both warm ( n = 1,519) and cold season ( n = 779) as compared to sheep carcass during the warm ( n = 511) and cold season ( n = 229). In contrast, coleopterans were more abundant on the sheep carcass during the warm season ( n = 391) and cold season ( n = 135) as compared to the pig carcass in both warm season ( n = 261) and cold season ( n = 114). In overall, more flies and beetles were collected on both sheep and pig carcasses during the warm season, and this further highlight that temperature influenced the observed difference in the abundance of collected arthropod between seasons., Competing Interests: The authors declare that they have no competing interests., (© 2021 Tembe et al.)

Prosobranch snails and adult Paramphistomoidea flukes were collected from water bodies and cattle abattoir located in Mpumalanga province of South Africa, respectively. The snails were identified based on morphological characters as well as the ITS-2 and 16S markers as Melanoides sp. and Tarebia granifera , respectively, and the Paramphistomoidea flukes were identified as Calicophoron microbothrium using the ITS-1/5.8S/ITS-2 marker. After confirming identification, the snails were bred to first filial generation (F1) under laboratory conditions. Ninety snails were randomly selected from the laboratory-bred F1 snails and 25 Melanoides sp. and 20 T. granifera were exposed to C. microbothrium miracidia, and the same numbers were maintained as non-exposed controls. Results showed that C. microbothrium successfully established in Melanoides sp. and produced cercariae, and the prepatent period recorded was 21 days. Three snails shed cercariae at day 21 postexposure (PE), and rediae and free cercariae were detected in the soft tissues of one snail on dissection at day 44 PE. The same fluke did not establish in T. granifera . Melanoides sp. started producing offspring at day 7 PE, and T. granifera at day 14 PE. In conclusion, our results showed that Melanoides sp. used in this study is a suitable intermediate host for C. microbothrium under experimental conditions, and given the wide distribution of this snail species, it is important to determine its role in the natural transmission of other Calicophoron species that have been reported in South Africa., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Malatji, Myende and Mukaratirwa.)

Fasciolosis is a food- and water-borne disease caused by digenean trematode species, Fasciola hepatica and F. gigantica . They are widely distributed and infect a wide range of definitive hosts, causing enormous economic loss due to reduced productivity in domestic ruminants. The two species have been previously reported to be co-endemic in KwaZulu-Natal and Mpumalanga provinces of South Africa. Hybridization between the two species has been reported elsewhere. Despite the overlap of the two species in two provinces, there has been no attempt to determine the presence of the intermediate forms or hybrids. Therefore, this study aimed at morphological and molecular characterization of Fasciola spp. collected from cattle slaughtered at abattoirs located in the two provinces of South Africa, where two species are endemic. A total of 71 fluke specimens were collected cattle from abattoirs in Enhlazeni and Nelspruit in Mpumalanga province and Pietermaritzburg in KwaZulu-Natal province of South Africa, and Zimbabwe (Bulawayo abattoir). Fasciola gigantica specimen collected from Zimbabwe where it has been confirmed as the only species occurring and this was used as control in the morphological and molecular assessment of the collected specimens. Of the 71 specimens collected, 37 were classified as F. hepatica , 12 as F. gigantica and 22 as Fasciola spp using morphological characters. Of these species, 11 of 37  F. hepatica and 6 of 22 Fasciola sp were found to be aspermic or having very scanty sperm. Fifteen flukes which were spermatic were all identified morphologically as F. gigantica whilst 5 flukes which were aspermic were identified morphologically as F. hepatica . Molecular analysis of the same 15 spermatic specimens confirmed the presence of F. hepatica ( n  = 9) and F. gigantica ( n  = 6) using the CO1 marker and as F. hepatica ( n  = 4), F. gigantica ( n  = 7) and Fasciola sp. ( n  = 1) for the same specimens using the ITS-1/5.8S/ITS-2 marker. The remaining 4 aspermic flukes (one did not resolve) were all identified morphologically as F. hepatica and molecular analysis confirmed them as F. hepatica ( n  = 4) by both CO1 and ITS-1/5.8S/ITS-2. Phylogenetic analysis based on both CO1 and ITS-1/5.8S/ITS-2 showed that F. hepatica species formed a moderately supported monophyletic clade with F. gigantica . Their ancestral history was further confirmed by haplotype network, which formed novel haplotypes that corresponded with the structure of the phylogenetic tree. Results from this study showed that morphological characters alone have limitations in identifying F. hepatica and F. gigantica in areas where the two species occur, although both methods confirmed the presence of F. gigantica occurring in Zimbabwe, F. hepatica in KwaZulu-Natal, and both species occurring in Mpumalanga province. Therefore, the use of morphological techniques, complemented by molecular techniques are recommended, especially in endemic areas where the two species are co-endemic., Competing Interests: None., (© 2021 The Authors. Published by Elsevier Inc. on behalf of International Association of Food and Waterborne Parasitology.)

Background: Giardia and Cryptosporidium species are significant zoonotic parasites of humans and domesticated animals., Objectives: The study aimed to determine the prevalence of Giardia and Cryptosporidium in livestock and dogs of the Magude District., Method: The flotation technique (Willis), modified Ziehl-Neelsen (mZN) and direct and indirect immunofluorescence (DIF and IIF) techniques were applied to determine the prevalence of Giardia and Cryptosporidium species in faecal samples of dog pups (156), goat kids (60) and calves (480) from the Magude District of Mozambique from February to September 2015., Results: Using Willis, IIF and DIF, the prevalence of Giardia in calves was 0%, 8.1%, and 6.0%; in dogs 0.6%, 8.3% and 5.7% and for goats 0% and 13.3% (IIF was not performed), respectively. The prevalence of Cryptosporidium in calves using Willis, mZN, IIF and DIF was 0%, 3.8%, 4.7% and 0.4% and in dogs 0%, 0.6%, 6.4% and 0.6%, respectively. The parasite was not detected in goats., Conclusion: Results from the present study showed that IIF performed better diagnosis of Giardia and Cryptosporidium, and that the mZN can be used as an alternative for Cryptosporidium because of the high cost of IIF. There is a need for identification of genotypes or subtypes of these parasites through application of molecular techniques in order to determine their zoonotic potential, and we advocate a 'one health' approach in the control and prevention of these parasites.

Lymnaeidae snail species are known to be intermediate hosts of human and livestock helminths parasites, especially Fasciola species. Identification of these species and their geographical distribution is important to better understand the epidemiology of the disease. Significant diversity has been observed in the shell morphology of snails from the Lymnaeidae family and the systematics within this family is still unclear, especially when the anatomical traits among various species have been found to be homogeneous. Although there are records of lymnaeid species of southern Africa based on shell morphology and controversial anatomical traits, there is paucity of information on the molecular identification and phylogenetic relationships of the different taxa. Therefore, this study aimed at identifying populations of Lymnaeidae snails from selected sites of the Okavango Delta (OKD) in Botswana, and sites located in the KwaZulu-Natal (KZN) and Mpumalanga (MP) provinces of South Africa using molecular techniques. Lymnaeidae snails were collected from 8 locations from the Okavango delta in Botswana, 9 from KZN and one from MP provinces and were identified based on phylogenetic analysis of the internal transcribed spacer (ITS-2). Analyses based on the ITS-2 marker identified the presence of a well-supported Radix clade containing Radix auricularia, R. natalensis and R. rubiginosa, which were not well resolved. Experimental samples from the OKD and KZN present in this clade were referable to these species. An unidentified experimental taxon from the OKD formed a well-supported sister clade to the Radix clade, although it was not possible to identify it. Galba truncatula was well supported in a sister relationship to a well-supported Pseudosuccinea columella clade which included samples from MP and KZN provinces of South Africa. We observed that P. columella shared the same habitats with R. natalensis and R. auricularia in KZN. Our study contributes new knowledge on the Lymnaeidae species present in Southern Africa and their phylogenetic relationships. The study further identifies the species which are likely to co-exist in the same environment and this information will be of use to those designing control programs for fasciolosis. This is the first study reporting the presence of R. auricularia in the OKD of Botswana and KZN province of South Africa., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A systematic review was conducted focusing on the distribution of Fasciola species and their snail intermediate hosts (IHs) in East and Southern Africa. The reviewed literature showed that both Fasciola hepatica and F. gigantica are present in East and Southern Africa, and infect a wide range of domestic and wild ruminants. Fasciola gigantica was reported in six East African and five Southern African countries, where Radix natalensis (found in low altitudes) was reported to be the main IH. Fasciola hepatica was reported in Tanzania and Ethiopia (East Africa), and in South Africa and Zimbabwe (Southern Africa), where Galba truncatula (found in high altitudes) was documented as the IH in all countries except in Zimbabwe. Both Fasciola species were documented in Tanzania, Ethiopia, Zimbabwe and South Africa. An overlap of the two was observed in areas with an intermediate altitude in Ethiopia and South Africa, where Pseudosuccinea columella was widespread and assumed to transmit both species. Pseudosuccinea columella has been reported in South Africa and Namibia, and proven to transmit F. gigantica in South Africa; its role in Namibia in the transmission of Fasciola species has not been reported. Other lymnaeid species such as R. rubiginosa were reported in South Africa, and R. auricularia in South Africa and Botswana; their role in the transmission of Fasciola species has not been proven. Future studies should aim to determine the role of P. columella in the geographical spread of the two species in East and Southern African countries.

The main intermediate host of Fasciola gigantica in sub-Saharan Africa is Lymnaea (Radix) natalensis. Lymnaea (Pseudosuccinea) columella is capable of transmitting both F. gigantica and F. hepatica and has been reported to be present in South Africa. To date, no natural infection with F. gigantica has been reported despite the wide distribution of the snail. The aim of this study was to confirm whether L. (P.) columella was transmitting F. gigantica and/or F. hepatica in selected locations of KwaZulu-Natal and Eastern Cape provinces of South Africa. Lymnaea (Pseudosuccinea) columella snails were collected from two locations in two provinces of South Africa and screened for cercariae shedding. This was followed by humanely sacrificing the screened snails, and whole tissue of each individual snail was homogenized and amplified using primers designed to amplify the ITS-1 region of Fasciola spp. No cercariae were shed from the screened snails and molecular analysis showed that snails from the two locations were infected with F. gigantica. This study confirms natural infection of L. (P.) columella with F. gigantica in South Africa, where F. gigantica and F. hepatica have already been reported to coexist. Although L. (P.) columella is able to transmit the two species, surprisingly no infection with F. hepatica was detected from the screened snails. The natural intermediate host of F. gigantica in southern Africa, including South Africa, is Lymnaea (Radix) natalensis and comparative studies are needed to determine the competence of the two snail species in the transmission of F. gigantica.

Trichinella species are widely distributed on all continents with the exception of Antarctica, although the full spectrum of Trichinella species found in sub-Saharan African countries, and their hosts, has not been fully documented. This study was conducted to determine the prevalence of Trichinella in wildlife from the Greater Kruger National Park (GKNP) and adjacent areas located in the Limpopo and Mpumalanga provinces of South Africa, and to identify the species and/or genotypes of Trichinella larvae isolated from muscle tissues, using molecular techniques. A review of Trichinella spp. and their wildlife hosts reported during 1964-2011 was also conducted and the results were compared with our current study. Ninety samples representing 15 mammalian, two bird and three reptile species were screened for Trichinella infection during 2012-2016, using artificial digestion. Isolates detected were identified using a multiplex polymerase chain reaction (PCR) amplification of the internal transcriber spacers ITS1 and ITS2, and expansion segment V (ESV) regions of ribosomal DNA, followed by molecular analysis of the sequences. Twenty samples from seven wildlife species were positive for Trichinella spp. larvae, with an overall prevalence of 21.1% (20/90). The prevalence was higher in carnivores (18.9%, 18/90) than in omnivores (2.2%, 2/90). Analysis of sequences showed that eight of the isolates - two from spotted hyaena (Crocuta crocuta) (2/8), three from lion (Panthera leo) (3/13), one from leopard (Panthera pardus) (1/6), one from small spotted genet (Genetta genetta) (1/2) and one Nile monitor lizard (Varanus niloticus) (1/2) - conformed to Trichinella zimbabwensis. One isolate from a hyaena was grouped under the encapsulated species clade comprising T. nelsoni and genotype Trichinella T8 reported to be present in South Africa. This is the first report confirming natural infection by T. zimbabwensis in hyaena, leopard, genet and Nile monitor lizard, adding to the body of knowledge on the epidemiology of Trichinella infections in the Greater Kruger National Park of South Africa. Ten Trichinella-like larval isolates recovered after digestion from four wildlife species in this study (2012-2016) revealed inconclusive results due to DNA degradation resulting from poor storage or too few larvae for analysis, in comparison to 20 unidentified isolates from five wildlife species during the 1964-2011 period.