Multidrug resistant enterohaemorrhagic Escherichia coli serogroups in the faeces of hunted Wildlife, Abeokuta, Nigeria.

Wildlife play significant roles in the dissemination and zoonotic transmission of pathogens. The enterohaemorrhagic Escherichia coli (EHEC) are associated with complicated cases of food-borne illnesses. This study investigated the presence of EHEC serogroups (O26, O45, O103, O145, O91, O111, O128, O121 and O157) in wildlife species: cane rats (Thryonomys swinderianus), royal antelope (Neotragus pygmaeus), African giant rats (Cricetomys gambianus) and waterbuck (Kobus ellipsiprymnus). EHEC and non-EHEC isolates from these wildlife sources were tested for susceptibility to antimicrobial agents. Overall, 127 (83.0 %) of 153 samples yielded E. coli. Nine (5.9%) samples were positive for EHEC belonging to three serogroups as follows; O26 (n=2), O111 (n=2) and O103 (n=5). The EHEC isolates were from cane rats (n=6) and royal antelope (n=3) and possessed virulence-associated genes stx1 (77.8%), stx2 (100.0%), eaeA (100.0%) and hlyA (100.0%). Overall, 127 E. coli isolates showed resistance to ampicillin (99.2%), ceftiofur (90.6%),  tetracycline (90.0%), cephalexin (87.4%), cefotaxime (50.4%), streptomycin 42.5%, ceftazidime (41.7%), nalidixic acid (37.0%),  ciprofloxacin (43.6%), amoxicillin/clavulanic acid (32.3%), gentamicin (27.6%), sulphamethoxazole/trimethoprim (25.2%), norfloxacin (17.3%) and chloramphenicol (11.0%). The roles of wildlife in the dissemination and transmission of antimicrobial resistant and zoonotic bacteria should not be neglected for effective preventive and control strategies.

Molecular detection of Ehrlichia ruminantium in engorged ablyomma variegatum and cattle in Ogun State, Nigeria.

Early diagnosis of Ehrlichia ruminantium in cattle is a recipe for effective control of heartwater in ruminants. Hence, we assessed the presence of E. ruminantium in the blood of cattle and the engorged Amblyomma variegatum by nested PCR. The electrophoresed PCR products obtained after primary and secondary amplifications revealed amplicon sizes of about350 bp and 280 bp respectively, which corresponded with the partial region of pSC20 gene amplified. Sequences obtained had 95-99% homology with those sequences available in GenBank. The prevalence of the E. ruminantium in ticks (50%; 126/252) was significantly (p < 0.05) higher than that in cattle blood 23.55% (61/259). The prevalence was significantly (p < 0.05) higher in ticks from adult cattle 51.47% (133/259) than those from the young cattle 44.86% (116/259) and in tick from females 54.55% (141/259) than in ticks from the males 41.38% (107/259). Alignment of autochthonous sequences revealed that the three sequences were polymorphic with two sequences showing similar nucleotides deletion at points 87-91 and 107-108. The phylogenetic trees inferred by ML showed topologies with two autochthonous sequences, one each from cattle blood and tick, clustering together in one clade and the other clustering within those sequences from South Africa and Zimbabwe in another clade. In conclusion, this study revealed a higher prevalence of E. ruminantium in engorged A. variegatum than in the blood of infected cattle. Hence, it is suggested that the amplification that targets the pCS20 gene in engorged ticks may be more suitable to determine the E. ruminantium carrier status of cattle.

Multi-locus genotyping and phylogenetic analyses of Giardia intestinalis isolates from indigenous goats in Ogun State, Nigeria.

Giardiasis is a common gastrointestinal disease of humans and various animal species worldwide. In this study, 302 stool samples were collected from West African Dwarf and Sokoto Red breeds of goats in Ogun State, Nigeria, and screened for Giardia intestinalis coproantigens using enzyme-linked immunosorbent assay (ELISA). The genotypes of G. intestinalis in faecal samples collected from 152 goats raised on selected farms were identified by polymerase chain reaction (PCR) amplification and sequence analyses of the small subunit ribosomal RNA (ssu rRNA), glutamate dehydrogenase (gdh), triosephosphate isomerase (tpi) and ß-giardin (bg) genes. Based on ELISA, an overall prevalence of 45.7% was recorded with the infection rates in pre-weaned (60.2%) and post-weaned goat kids (51.5%) being significantly (p < 0.05) higher than in adults (28.2%). Giardia intestinalis DNA was amplified in 31.6% and 29.6% of goat faeces at the ssu rRNA and gdh loci respectively. These were genotyped at the ssu rRNA locus as assemblages B (n = 13) and E (n = 36). Similar results were observed at the gdh locus except that eight isolates contained assemblage E mixed with either assemblage A or B. Additionally, sub-assemblages BI (n = 7) and BIII (n = 2) were identified with up to four single nucleotide polymorphisms (SNPs) occurring in these isolates. Multilocus genotypes (MLG) of all assemblage E isolates were identified using the ssu rRNA and gdh loci while MLG of all isolates containing assemblage B and mixed assemblages were determined after further typing at the tpi and bg loci. Forty-two MLG isolates were identified and these comprised 32, 8 and 2 (sub)-assemblage E, BI and BIII respectively. All isolates with mixed assemblages at the gdh locus were consequently designated as assemblage E by MLG. The assemblage E isolates from goats were genetically related to isolates from cattle, sheep and goats while the assemblage B isolates were related to isolates of human, pig and lemur origin. This suggests that G. intestinalis isolated from goats bred in Ogun State, Nigeria may be capable of cross-species transmission and may be of zoonotic importance.

Prevalence and multilocus genotypes of Giardia duodenalis infecting pigs in Ogun state, Nigeria.

Giardia duodenalis is an intestinal flagellated protozoan parasite that is infectious to humans and a wide range of animals worldwide. While varying prevalence rates have been reported in pigs worldwide, there are currently no published reports on the genotypes of Giardia infecting pigs in any African country. The present study is on the prevalence and genotypes of G. duodenalis in 209 pigs raised on four farms in Ogun State Nigeria. Using an enzyme-linked immunosorbent assay (ELISA) kit, Giardia duodenalis coproantigens were detected on all farms and in 25.4% (53/209) of pigs sampled. However, there was no significant influence (p > 0.05) of age, sex and stool consistencies of the pigs on the distribution of the infection. Genotyping of Giardia duodenalis in all ELISA-positive samples, achieved by the amplification of the small subunit ribosomal RNA (ssu rRNA), glutamate dehydrogenase (gdh), triosephosphate isomerase (tpi) and beta giardin (bg) genes, identified 14 and 37 assemblage B and E isolates respectively while mixed infection by both assemblages was recorded in two isolates. Novel nucleotide substitutions were identified in four assemblage B isolates at the ssu rRNA locus. Genetic diversity was observed among the assemblage B isolates after multiple alignment analyses of the gdh, tpi and bg sequences whereby sub-assemblages BII (n = 2), BIII (n = 9) and BIV (n = 3) were identified. The assemblage B isolates from pigs in this study were phylogenetically related to isolates from humans, marmoset and cattle while the assemblage E isolates were related to isolates from sheep, goats and cattle. These findings suggest that pigs in southwest Nigeria predominantly harbour G. duodenalis isolates that could be infectious to other animal species and to a lesser extent, isolates that may be of zoonotic importance.

Multilocus genotyping of Giardia duodenalis infecting rabbits in Ogun State, Nigeria.

Giardiasis is a cosmopolitan gastrointestinal protozoal parasite that infects humans and various animals worldwide. To assess the zoonotic transmission potential of Giardia, molecular characterization is required. We are unaware of any report on the genotypes of Giardia infecting rabbits in Nigeria. Molecular detection and genotyping of Giardia duodenalis were conducted in a herd of adult Chinchilla rabbits (Oryctolagus cuniculus) managed on the Teaching and Research farm of the Federal University of Agriculture, Abeokuta located in a southwestern state of Nigeria by analysis of the small-subunit ribosomal RNA (ssu rRNA), glutamate dehydrogenase (gdh), triosephosphate isomerase (tpi) and beta-giardin (bg) genes. An overall prevalence of 72.3% (60/83) was recorded in the rabbits with no statistically significant (p > .05) influence of sex on the distribution of the infection in the herd. All the 19 isolates amplified at the four genetic loci were identified as G. duodenalis assemblage BIV by multiple alignment analysis of their consensus sequences. Novel nucleotide substitutions were identified in two isolates at the ssu rRNA locus. Phylogenetic analysis revealed that all ssu rRNA genotypes were closely related to G. duodenalis assemblage B of cattle and human origin. Findings of this study suggest that the rabbits harbour potentially zoonotic assemblage BIV that portends a high risk to students and staff of the University who are in regular contact with the animals.

Reliability of clinical monitoring for the diagnosis of babesiosis in dogs in Nigeria.

Babesiosis accounts for a high percentage of hospital cases in canines in Africa, with about 40% mortality in the cases presented. In Nigeria, records show an estimated 30% annual morbidity when diagnosis is largely based on clinical and laboratory findings. This study monitored clinical indices associated with canine babesiosis. One hundred and three babesiosis-suspected dogs were selected on the basis of clinical signs of anorexia, fever, presence of ticks, and enlarged lymph nodes or spleen when clinical parameters were recorded at the time of presentation. Parasite detection was done using thin blood smears; that is, the presence of Babesia merozoites was compared between capillary and cephalic blood. Blood was also assayed for hematology and blood chemistry using automated blood analyzers. The babesiosis-infected dogs' outcome was monitored. Data obtained were analyzed using chi-square test, analysis of variance, and Pearson's correlation. Results based on thin blood smears showed that 61.1% of the dogs were positive for Babesia species. Breed disposition, sex, and age did not significantly influence the incidence of Babesia canis, while mean rectal temperatures did not differ significantly between the cases (P>0.05). Heart rate and pulse rates of Babesia-positive dogs were significantly (P<0.05) higher than those that were negative. The packed cell volume between the cases was not significantly different, with the values in the positive and negative case obtained being 26.4% ±11.26% and 31.6%±11.9%, respectively, with a range of 6% to 50% and 10% to 47% observed, respectively. Normal leukogram was also observed in 62% of the Babesia-positive cases while 22.2% and 15.8% had leukocytosis and leukopenia, respectively. Most of the positive cases whose results were based on thin blood smear were treated with 5% oxytetracycline for 5 days and fully recovered. Pearson's correlation was used to give relationship in the observed data. This study concluded that clinical indices are not reliable markers in the diagnosis of canine babesiosis.

Implications of Ebola virus disease on wildlife conservation in Nigeria.

The recent Ebola Virus Disease outbreak in some West African countries spanning from late 2013 and currently on as of 13th March, 2015 is the most widespread and fatal with human mortality that has surpassed all previous outbreaks. The outbreak has had its toll on conservation of endangered species. This portends danger for the wild fauna of the country if proactive measures are not taken to prepare grounds for evidence-based assertions concerning the involvement of wild species. To this end, there is an urgent need for sweeping census of reserves, national parks and wetlands. As well as the creation of a system involving reportage by sectors like the industries (extractive and construction) including persons and organisations involved with wildlife related activities. This documentation of die offs and unusual events to collaborating institutions, will help in monitoring trends which hitherto would have gone unnoticed. The importance of bats and primates in agriculture and public health via consumption of vermin insects and seed dispersal cannot be over-emphasized. There is the need for caution on the tendencies to destroy indicator species which could be silent pointers to emerging or re-emerging health and environmental issues. Wildlife resources are still reliably useful and caution is advised in the use of blanket destructive policies like fumigation of caves, indiscriminate culling and poisoned baits to destroy supposedly Ebola Disease Virus wildlife reservoirs. This paper highlights the immediate conservation problems and likely future implications of Ebola saga in Nigeria. It tries to identify the gaps in wildlife researches and makes recommendations for probable workable conservation strategies.