Molecular detection of Bartonella quintana, Acinetobacter baumannii and Acinetobacter haemolyticus in Pediculus humanus lice in Nigeria, West Africa.

The human lice Pediculus humanus is distributed worldwide but, it thrives and flourishes under conflict situations where people are forced to live in crowded unhygienic conditions. Molecular methods were used to identify and screen human lice for the DNA of pathogens of public health importance in an area that has been under insurgency related to religious and political conflicts with tens of thousands of internally displaced people (IDP). DNA of Bartonella quintana, Acinetobacter baumannii and Acinetobacter haemolyticus was detected in 18.3%, 40.0% and 1.7%, respectively, of human lice collected from children in Maiduguri, Nigeria. More body lice than head lice were positive for pathogen's DNA (64.3% vs. 44.4%; χ2 = 1.3, p = 0.33), but the difference was not significant. Two lice samples were found to harbour mixed DNA of B. quintana and A. baumannii. Phylogenetic analysis of the cytochrome b (cytb) gene sequences of the positive lice specimens placed them into clades A and E. This is the first report on the molecular identification of human lice and the detection of the DNA of pathogens of public health importance in lice in Nigeria, West Africa. The findings of this study will assist policy makers and medical practitioners in formulating a holistic healthcare delivery to IDPs.

Phylogenetic and network analysis of Pediculus humanus in Nigeria reveal the presences of clade E body lice and novel haplotypes.

An investigation was conducted for the first time to determine the prevalence and genetic diversity of human lice, for the first time in Nigeria, using conventional PCR and sequencing methods. Three mitochondrial genes, cytochrome oxidase subunit 1 (cox1), cytochrome b (cytb), and 12S rRNA of Nigerian human lice, were amplified, sequenced, and analyzed. Overall, high prevalence (72.5%; 103/142) of lice infestation was recorded among the examined volunteers. Head lice infestation was more common 63 (61.2%) than body lice infestation 34 (33.0%). Co-infestation with both head and body lice was recorded in six humans (5.8%). The Nigerian human lice specimens were placed mostly into clade A with few in clade E, including body lice for the first time. Six, three, and eight haplotypes of Nigerian human lice were obtained for the cytb, cox1, and 12S rRNA genes, respectively. Additionally, one (E51), three (A31, A32, and E5), and six (A20, A21, A23, A24, A30, and E1) novel haplotypes were recorded for cox1, cytb, and 12S rRNA, respectively, from the Nigerian specimens which were corroborated by the ML phylogenetic trees and MJ network analyses. Genetic diversity indices indicate minimal variation in the parameters analyzed among the clades of the three genes. However, a statistically significant Snn test, negative Tajima's D test for clade A (cox1 and 12S rRNA genes), and negative Fu and Li's D test in clade A for cox1 gene indicate a geographical structure and the signature of population expansion of the Nigerian human lice. The findings from this study provide additional data on the human lice structure in Africa.

Genomic characterization of Alphacoronavirus from Mops condylurus bats in Nigeria.

Coronaviruses (CoVs) are responsible for sporadic, epidemic and pandemic respiratory diseases worldwide. Bats have been identified as the reservoir for CoVs. To increase the number of complete coronavirus genomes in Africa and to comprehend the molecular epidemiology of bat Alphacoronaviruses (AlphaCoVs), we used deep metagenomics shotgun sequencing to obtain three (3) near-complete genomes of AlphaCoVs from Mops condylurus (Angolan free-tailed) bat in Nigeria. Phylogenetic and pairwise identity analysis of open reading frame 1ab (ORF1ab), spike (S), envelope (E), membrane (M) and nucleocapsid (N) genes of AlphaCoV in this study to previously described AlphaCoVs subgenera showed that the Nigerian AlphaCoVs may be members of potentially unique AlphaCoV subgenera circulating exclusively in bats in the Molossidae bat family. Recombination events were detected, suggesting the evolution of AlphaCoVs within the Molossidae family. The pairwise identity of the S gene in this study and previously published S gene sequences of other AlphaCoVs indicate that the Nigerian strains may have a genetically unique spike protein that is distantly related to other AlphaCoVs. Variations involving non-polar to polar amino acid substitution in both the Heptad Repeat (HR) regions 1 and 2 were observed. Further monitoring of bats to understand the host receptor use requirements of CoVs and interspecies CoV transmission in Africa is necessary to identify and prevent the potential danger that bat CoVs pose to public health.

Mycoplasma wenyonii and Candidatus Mycoplasma Haemobos in Pastoralists Cattle in Nigeria.

PURPOSE: The extensive migration practiced by pastoralists cattle exposes them to a variety of pathogens and vectors which may sometimes lead to severe disease outcomes. Moreover, the synergistic effect of multiple parasitism on the productivity of livestock has been well recognized. This is particularly true where the livestock production system predisposes the animals to constant and heavy infestation with arthropod vectors. METHODS: The presences, prevalence and risk factors for hemotropic Mycoplasma (hemoplasma) infection in cattle in Nigeria was investigated using a PCR and sequencing approach. DNA, extracted from 566 cattle blood samples, collected from 10 states from the three agro-ecological zones (AEZs) of Nigeria, from April 2021 to March 2022, were screened for the presences of hemotropic Mycoplasma spp. DNA. RESULTS: The DNA of hemoplasmas was detected in 48 out of the 566 (8.5%) samples, 12 (25%) of them were identified as Mycoplasma wenyonii and 19 (38.6%) as 'Candidatus Mycoplasma haemobos'. Coinfection with both species was detected in 17 (35.4%) of the samples. High prevalence and risk of hemoplasmas infection was associated with sex of the cattle (bulls were more affected; p = 0.005) and the packed cell volume (p = 0.009), but not with the age (p = 0.08), breed (p = 0.22), body condition (p = 0.052), source of the samples (p = 0.45) or the AEZs (0.59). This is the first nationwide survey of hemotropic mycoplasmas in cattle in Nigeria using this molecular approach. CONCLUSION: Further studies to determine the veterinary and public health significance of these pathogens, which were previously associated with varying degrees of clinical signs and production losses, are recommended in Nigerian cattle.

Molecular detection of Theileria annulata, Theileria mutans and Theileria velifera but no evidence of Theileria parva infected or vaccinated cattle in Nigeria despite extensive transboundary migrations.

The extensive livestock management system predominant in Nigeria necessitates active disease surveillance for the early detection and prompt control of transboundary animal diseases. Theileriae are obligate intracellular protozoa which infect both wild and domestic bovidae throughout much of the world causing East Coast Fever (Theileria parva), Tropical or Mediterranean theileriosis (Theileria annulata) or benign theileriosis (Theileria mutans; Theileria velifera). This study aimed to detect and characterize Theileria spp. infecting cattle in Nigeria using conventional PCR and sequencing approach. Five hundred and twenty-two DNA samples obtained from different cattle blood samples were subjected to PCR targeting the 18S rRNA gene of piroplasmida and specifically, the p104 kDa and Tp1 genes for the evidence of infection or vaccination respectively, with T. parva. A total of 269 out of 522 (51.5%) of the cattle tested PCR- positive for DNA of piroplasmida. Nucleotide sequence and phylogenetic analyses showed that the cattle were infected with T. annulata, T. mutans and T. velifera. Piroplasmida DNA was associated with sex (ꭓ2 = 7.2; p = 0.007), breed (ꭓ2 = 115; p = 0.000002) of animals and the state where the samples were collected (ꭓ2 = 78.8; p = 0.000002). None of the samples tested positive for T. parva DNA or showed evidence of vaccination (Tp1 gene). This is the first report on the molecular detection and characterization of T. annulata in the blood of cattle from Nigeria. Continuous surveillance of Nigerian cattle for East Coast Fever (ECF) is encouraged considering the recent report of the disease in cattle in the neighboring country, Cameroon, where unregulated transboundary cattle movement into Nigeria has been observed.

Rickettsia africae and Rickettsia massiliae in ixodid ticks infesting small ruminants in agro-pastoral settlements in Plateau State, Nigeria.

Arthropods, especially ixodid ticks, have been incriminated in the epidemiology of Spotted Fever Group rickettsioses globally leading to an increasing spectrum of emerging and re-emerging zoonoses with attendant consequences on trade and tourism. The objective of this study was to determine the role of ixodid ticks infesting small ruminants in Plateau State, Nigeria, in the epidemiology of Spotted Fever Group Rickettsiae (SFGR) in the study area. DNA from 130 out of 323 ixodid ticks collected from 179 goats and 121 sheep owned by agro-pastoralists in Plateau State were screened for the evidence of SFGR by molecular methods. Six tick species from four genera were identified: Amblyomma, Hyalomma, Rhipicephalus (Boophilus) and Rhipicephalus. Rhipicephalus sanguineus sensu lato (s.l.) was the predominant (54.5%) species among collected ticks. Tick infestation was significantly associated with the species of small ruminants, the sex of the animals and the sampling locations except for Jos South. Conventional PCR targeting the 381 bp of the citrate synthase (gltA) and 820 bp of the outer membrane protein B (ompB) genes detected DNA of SFGR in nine and eight samples, respectively. Sequence analysis revealed that five sequences obtained from Amblyomma variegatum were 99-100% identical to Rickettsia africae and three sequences from Rh. sanguineus (s.l.) were 100% identical to Rickettsia massiliae reported from Spain. To our knowledge, this is the first report of the detection of R. africae DNA in Am. variegatum collected from small ruminants in Plateau State. Ixodid ticks infesting small ruminants in Plateau state harbor DNA of SFGR with potential veterinary and public health implications.

Straw-Colored Fruit Bats (Eidolon helvum) and Their Bat Flies (Cyclopodia greefi) in Nigeria Host Viruses with Multifarious Modes of Transmission.

Background: Bat flies (Diptera: Hippoboscoidea: Nycteribiidae and Streblidae) are increasingly appreciated as hosts of "bat-associated" viruses. We studied straw-colored fruit bats (Eidolon helvum) and their nycteribiid bat flies (Cyclopodia greefi) in Nigeria to investigate the role of bat flies in vectoring or maintaining viruses. Methods: We captured bats and bat flies across northern Nigeria. We used metagenomics to identify viruses in 40 paired samples (20 flies from 20 bats). We characterized viruses using genomic and phylogenetic methods, and we compared infection frequencies in bats and their bat flies. Results: In 20 bats, we detected two individuals (10%) infected with eidolon helvum parvovirus 1 (BtPAR4) (Parvoviridae; Tetraparvovirus), previously described in Ghana, and 10 bats (50%) with a novel parvovirus in the genus Amdoparvovirus (Parvoviridae). The amdoparvoviruses include Aleutian disease virus of mink and viruses of other carnivores but have not previously been identified in bats or in Africa. In 20 paired bat flies (each fly from 1 bat) all (100%) were infected with a novel virus in the genus Sigmavirus (Rhabdoviridae). The sigmaviruses include vertically transmitted viruses of dipterans. We did not detect BtPAR4 in any bat flies, and we did not detect the novel sigmavirus in any bats. However, we did detect the novel amdoparvovirus in 3 out of 20 bat flies sampled (15%), including in 2 bat flies from bats in which we did not detect this virus. Discussion: Our results show that bats and their bat flies harbor some viruses that are specific to mammals and insects, respectively, and other viruses that may transmit between bats and arthropods. Our results also greatly expand the geographic and host range of the amdoparvoviruses and suggest that some could be transmitted by arthropods. Bat flies may serve as biological vectors, mechanical vectors, or maintenance hosts for "bat-associated" viruses.

Detection of Alpha- and Betacoronaviruses in Frugivorous and Insectivorous Bats in Nigeria.

The rise of bat-associated zoonotic viruses necessitates a close monitoring of their natural hosts. Since the detection of severe acute respiratory syndrome coronavirus (SARS-CoV), it is evident that bats are vital reservoirs of coronaviruses (CoVs). In this study, we investigated the presence of CoVs in multiple bat species in Nigeria to identify viruses in bats at high-risk human contact interfaces. Four hundred and nine bats comprising four bat species close to human habitats were individually sampled from five states in Nigeria between 2019 and 2021. Coronavirus detection was done using broadly reactive consensus PCR primers targeting the RNA-dependent RNA polymerase (RdRp) gene of CoVs. Coronavirus RNA was detected in 39 samples (9.5%, CI 95%: [7.0, 12.8]), of which 29 were successfully sequenced. The identified CoVs in Nigerian bats were from the unclassified African alphacoronavirus lineage and betacoronavirus lineage D (Nobecovirus), with one sample from Hipposideros ruber coinfected with alphacoronavirus and betacoronavirus. Different bat species roosting in similar or other places had CoVs from the same genetic lineage. The phylogenetic and evolutionary dynamics data indicated a high CoV diversity in Nigeria, while host switching may have contributed to CoV evolution. Robust sentinel surveillance is recommended to enhance our knowledge of emerging and re-emerging coronaviruses.

Absence of DNA and anti-leishmanial antibodies in dogs (Canis familiaris) in Plateau state, Nigeria.

Dogs are important sentinels for the surveillance of some zoonotic diseases including human leishmaniasis. To obtain information on the role of dogs in the epidemiology of leishmaniasis in Nigeria, 98 sera and 204 DNA samples obtained from dogs were screened for anti-leishmania antibodies and DNA of Leishmania spp. using the enzyme linked immunosorbent assay (ELISA) and PCR, respectively. Initially, three out of the 98 sera samples had ELISA borderline optical density (OD) values and were retested. Two out of the three samples turned out to be negative while one sample gave yet a borderline OD value on a retest. A real time polymerase chain reaction (RT-PCR) targeting the 120-bp fragment of the minicircle kDNA of Leishmania spp. run on DNA extracted from EDTA preserved blood of the borderline positive serum failed to amplify the 120-bp sequence of Leishmania spp. In the second phase of the study, 204 DNA from dog blood samples were subjected to conventional PCR targeting the 300-350 bp of the internal transcribe spacer region 1 (ITS1) of Leishmania spp. None of the samples could be amplified (n = 204, 0%). Our study suggests that L. infantum is not prevalence in Jos, Plateau State, Nigeria and this should be confirmed using a larger sample of local dogs tested using PCR methods in lymphoid tissue samples.

Nucleotide sequence types (ntSTs) of Anaplasma marginale in cattle in Nigeria based on the major surface protein 5 (msp5) gene.

Bovine anaplasmosis caused by Anaplasma marginale is an important endemic disease that exerts negative impact on livestock production with huge socioeconomic consequences in most developing countries. Genetic studies have reported the existence of diverse ntSTs of A. marginale with varying pathogenicity in different countries. Continuous studies to obtain accurate information on disease etiologies is desirable for the formulation of cost-effective control measures. To this end, 582 blood samples from cattle were collected from 10 out of the 36 States of Nigeria from April 2021 to March 2022 and analyzed based on the major surface protein 5 (msp5) gene to determine the ntSTs of A. marginale in Nigeria. In all, 38 out of the 582 samples (6.5%) from cattle in the different Agro-ecological Zones (AEZs) of Nigeria were positive. The Nigerian A. marginale nucleotide sequences were 96.7 to 100% identical to sequences from other countries and were placed in distinct clusters with other A. marginale sequences deposited in GenBank. Network analysis revealed three ntSTs (#2, #4 & #8) of A. marginale from Nigeria with a nucleotide sequence type diversity (Hd) of 0.8, nucleotide diversity (Pi) of 0.015 and average number of nucleotide differences (k) of 7.09. Two different amino acid substitution sites were found in Nigerian and worldwide sequences at positions 148 and 160. This is the first nationwide report on the ntST diversity and genetic characterization of A. marginale in cattle in Nigeria based on the msp5 gene. Bovine anaplasmosis is widespread in Nigeria and deserves further attention.

Molecular detection and genetic characterization of Anaplasma marginale and Anaplasma platys in cattle in Nigeria.

Bovine anaplasmosis poses serious challenge to profitable livestock production in the tropics. Accurate information on the prevalence, distribution and genetic characteristics of Anaplasma spp. infections of cattle is invaluable for the design of cost-effective control measures. Blood samples from 275 cattle in Nigeria were screened for the DNA of Anaplasma spp. using species-specific primers and nucleotide sequence analysis. The DNA of Anaplasmataceae was detected based on 16S rRNA gene in 135 out of the 275 (49.1%) individuals examined, with 31 (23.0%) and 21(15.6%) being positive for Anaplasma marginale based on msp4 and msp2 genes, respectively. DNA of Anaplasma platys was detected in 62 (45.9%) based on groEL gene and in 27 (20.0%) using the A. platys species-specific primers. Presence of Anaplasma spp. DNA was significantly associated (p = 0.011) with the breed of the animals. Anaplasma nucleotide sequences of one group of the infected samples showed high identities of 99.0 to 100% (16S rRNA gene) and 99.6% (groEL gene) with reference sequences of A. platys, while those of another group matched to A. marginale references (msp2 with 98.9% and msp4 with 99.1%). Furthermore, phylogenetic analysis clustered the nucleotide sequences in this study with A. platys and A. marginale sequences in GenBank, confirming these relationships. For the first time, this study revealed the presence of mixed haplotypes in both A. platys and A. marginale in cattle in Nigeria. More studies are needed to elucidate the epidemiology and veterinary and public health significance of Anaplasma spp. infections in cattle in Nigeria.

Molecular identification and genetic analysis of Rhipicephalus sanguineus sensu lato of dogs in Nigeria, West Africa.

The brown dog tick, Rhipicephalus sanguineus sensu lato, is a ubiquitous and taxonomically controversial pest of dogs with immense veterinary and public health significance. Genetic analyses of specimens from various geographical origins reveal intraspecific diversity within the taxon. Little information is available on the genetic characteristics of R. sanguineus s.l. in Nigeria, West Africa. In this study, 460 bp of the mitochondrial 16S rDNA gene of R. sanguineus s.l. collected from dogs in different ecological zones of Nigeria was amplified, sequenced and characterized. Phylogenetic and pairwise analyses were used to compare the sequences generated in this study to each other and to sequences in GenBank. The sequences in this study were highly similar (>98%) to each other and clustered with sequences of the R. sanguineus s.l. tropical lineage in GenBank. None of the sequences in this study clustered with the 'southeastern Europe' or temperate lineage. The mean intraspecific divergence among R. sanguineus s.l. in this study was 1.7% (range: 0-8.0%). Furthermore, the sequences in this study showed mean divergence of 1.5% (0-10%), 5.0% (3.8-13.9%) and 9.7% (6.9-19.8%) from sequences of the tropical, southeastern Europe and temperate lineages, respectively. Interestingly, sequences in this study showed a mean divergence of 9.3% (1.0-17.8%) from the Rhipicephalus sp. morphotype 4 (GenBank acc. nr. KC243850) earlier identified from cattle in Nigeria, suggesting diversity in this taxon in Nigeria. Further studies are needed to elucidate the veterinary and public health significance of R. sanguineus s.l. in Nigeria taking into cognizance the existence of intraspecific variation in vector competence.

Seroprevalence of Spotted Fever Group Rickettsiae in extensively managed sheep and goats in Nigeria, West Africa.

Sheep and goats raised extensively are frequently infested by Ixodid ticks that may act as vectors or reservoirs of Spotted Fever Group Rickettsiae (SFGR). A study to determine the seroprevalence of SFGR infection in 300 sheep and goats in Plateau State, Nigeria was conducted from September to November, 2018 using the Indirect Fluorescence Antibody Test (IFAT). Overall, 85 out of 300 animals (28.3%) were seropositive to SFGR. Relatively higher seroprevalence was recorded in sheep than goats (28.8% vs 28.0%) but the difference was not statistically significant (p > 0.05). Furthermore, seropositivity was not affected by age, sex or location of the animals screened in this study. This is the first serological study to report the prevalence of SFGR in sheep and goats using IFAT in this study area. The presence of SFGR antibodies in domestic ruminants is of public health concern considering the close association between farmers and their animals occasioned by the management system practiced in the study area. This finding calls for further studies to evaluate the level of human exposure to this group of pathogen.

Molecular detection and characterization of pathogenic Leptospira species in bats (Chiroptera) roosting in human habitats in Nigeria, West Africa.

Leptospirosis is a neglected zoonosis with a nearly global distribution. In order to determine the role of bats in the epidemiology of leptospirosis in Nigeria, a total of 231 bats belonging to three families, Pteropodidae (n = 117), Molossidae (n = 107) and Nycteridae (n = 17), roosting in human habitats were screened by PCR and sequencing for the detection of pathogenic Leptospira species. DNA extracted from the kidneys of bats were subjected to conventional PCR targeting the rrs1, rrs2, flaB and secY genes for the detection of pathogenic Leptospira spp. Overall, 27 out of the 231 (11.7%) of the samples screened were positive for Leptospira spp. High prevalence (>80%) of Leptospira spp. DNA was detected in Chaerophon and Nycteris bat species captures in an abandoned well located within a human habitation. Sequences generated in this study were highly identical to Leptospira borgpetersenii and Leptospira interrogans and clustered with sequences of pathogenic species in GenBank. The detection of pathogenic Leptospira spp. was significantly associated (p < .001) with the bat species, feeding habit, roosting site and study location. To the best of our knowledge, this is the first molecular detection and characterization of pathogenic Leptospira spp. in bats from Nigeria. Results show that bats in Nigeria are infected with diverse Leptospira genotypes phylogenetically related to known pathogenic, including zoonotic taxa. Together, these findings reinforce bats' roles as potential reservoirs of Leptospira spp. and should be considered as a starting point for future comparative studies to improve our understanding of the epidemiology of this bacterial pathogen in Nigeria.

Seroprevalence of Theileria equi, Babesia caballi, and Trichinella spp. infections in horses and donkeys from Nigeria, West Africa.

The protozoan parasites Theileria equi and Babesia caballi, transmitted by ticks, cause equine piroplasmosis, the most prevalent tick-borne disease in equids. Trichinellosis is a worldwide food-borne zoonosis caused by helminth Trichinella spp. that can lead to serious disease in humans, with fatal outcome. Although the infection is rare in horses, it deserves attention due to the increasing use of horse meat as a source of protein for humans. Horse trichinellosis is caused by several Trichinella species, most commonly by T. spiralis. The aim of the study was to determine the prevalence of antibodies to T. equi, B. caballi and Trichinella spp. in equids from three states of Northern Nigeria. Serum samples were collected from 139 clinically healthy animals, comprising 115 horses and 24 donkeys. Antibodies to T. equi and B. caballi were detected in serum by competitive-inhibition enzyme-linked immunosorbent assay (cELISA) and antibodies to Trichinella spp. by ELISA. Antibodies to T. equi were detected in 34% of equids (41% horses and 0% donkeys), antibodies to B. caballi in 9% of equids (8% horses and 13% donkeys), and antibodies to Trichinella spp. in 4% of equids (4% horses and 0% donkeys). There was co-infection of T. equi and B. caballi in 1% of horses and co-infection of T. equi and Trichinella spp. in 2.6% of horses. This is the first report on seroprevalence of Trichinella spp. in equids from Northern Nigeria.

Canine gastrointestinal parasites as a potential source of zoonotic infections in Nigeria: A nationwide survey.

Dog feces may contain zoonotic parasites that contaminate the environment and serve as a potential source of infection to animals and humans. In this study, microscopic and molecular analyses were used to estimate the prevalence and intensity of gastrointestinal (GI) parasites and assess the risk factors for infection in 948 dogs in three climatically distinct zones of Nigeria. Zoonotic helminths including Strongyloides stercoralis, Ancylostoma braziliense, A. caninum and Toxocara canis were detected either as single or multiple infections in 377 (39.8 %) of dogs examined. At multiple logistic regression analyses, association was found between GI parasite infection and deworming practices and dog management. Regarding A. braziliense, A. caninum and T. canis infections, intensity of egg shedding was statistically associated with the age of the dogs and not with their sex or breed. The majority of GI parasite-positive dogs did not receive regular deworming treatment (59 %) and roamed freely (56 %) thereby constituting public health risk. This is the first nationwide survey and analyses of risk factors of GI parasites of dogs using molecular methods as confirmation of their identity. The zoonotic potential of these parasites is exacerbated by the lack of both operational national policies to control the population of free-roaming dogs and to promote responsible dog ownership, and veterinary public health programs for dogs.

Molecular evidence indicts Haemaphysalis leachi (Acari: Ixodidae) as the vector of Babesia rossi in dogs in Nigeria, West Africa.

Babesia rossi is the most pathogenic among the large canine babesias and it is the major cause of canine babesisosis in Nigeria. In South Africa it is transmitted by Haemaphysalis elliptica however, its putative vector in Nigeria where Rhipicephalus sanguineus is the most prevalent tick on dogs compared to Haemaphysalis species has not been ascertained. The incongruity between tick distribution and the frequent detection of B. rossi in Nigeria motivated this investigation to identify the local vector(s) of B. rossi. A total of 3805 ticks were collected from 363 naturally infested dogs from different parts of Nigeria. Of these numbers, 758 engorged female ticks; Rh. sanguineus (n = 660) and H. leachi (n = 98) were incubated for oviposition and hatching. After the completion of egg laying, Rh. sanguineus (n = 69) and H. leachi (n = 24) and their resulting progenies were screened for the presence of B. rossi DNA using a nested PCR targeting the 693 bp of the 18S rRNA gene of Babesia spp. Amplification and sequencing of B. rossi DNA was successful in the adults of H. leachi and their resulting egg and larval progenies but not in the adult Rh. sanguineus and progenies. The B. rossi DNA sequences from the H. leachi and their progenies have 99-100 % identity to each other and 98-99 % identical to sequences of B. rossi in GenBank (GenBank: MH143395.1), thus confirming transovarian passage. This evidence confirms for the first time following the reclassification of H. leachi to H. elliptica in South Africa the role of H. leachi in the transmission of B. rossi in dogs in Nigeria.

Field application of a novel multiplex qPCR assay reveals the occurrence of the zoonotic hookworm Ancylostoma braziliense in Nigerian dogs.

A number of gastrointestinal parasites have been reported to infect dogs in Nigeria, some of which have zoonotic potential. Of these, hookworms are the most prevalent, with both Ancylostoma caninum and Uncinaria stenocephala reported in the country. In this study, we subjected 203 hookworm microscopy-positive samples of the 885 individual faecal samples collected from dogs in Nigeria to a recently developed multiplex qPCR for the detection and characterisation of canine hookworm species. The qPCR demonstrated a diagnostic sensitivity of 98% (95% CI 95-99.4) allowing the detection of A. caninum and A. braziliense in 81.3% (165/203, 95% CI 75.3-86.1) and 51.2% (104/203, 95% CI 44.4-58) of the microscopy-positive faecal samples of dogs from Nigeria, respectively and 34.5% (70/203, 95% CI 28.3-41.3) of mixed infections with both hookworm species. The finding of A. braziliense is particularly worrisome given this is a well-known agent of persistent cutaneous larva migrans, commonly referred to as "creeping eruptions" in humans. Although this parasite has been diagnosed in locals and in people travelling in Nigeria suffering from dermatological illnesses, this represents the first molecular identification of A. braziliense in its canine reservoir in the country. These results update the occurrence and distribution of hookworm species affecting dogs in Nigeria highlighting the suitability of the newly developed multiplex qPCR assay as a high-throughput tool for the surveillance of zoonotic hookworms, globally.