Occurrence, antimicrobial resistance and whole genome sequence analysis of Salmonella serovars from pig farms in Ilorin, North-central Nigeria.

Salmonella enterica is a foodborne pathogen of global public health importance with developing countries mostly affected. Foodborne outbreaks are often attributed to pork consumption and Salmonella contamination of retail pork is directly linked to the Salmonella prevalence on farm. The widespread use of antimicrobials at different steps of swine production can favor resistant strains of Salmonella. The objectives of this study are to characterize the distribution, multilocus sequence typing (MLST), plasmid, virulence profiles and antimicrobial resistance of Salmonella serovars circulating in selected pig farms. Six hundred fecal samples were randomly collected from nine selected farms in Ilorin, Nigeria. Isolates were analyzed by cultural isolation using selective media, conventional biochemical characterization, serotyping, MLST and whole genome sequencing (WGS). Sixteen samples were positive for Salmonella sub-species, comprising of nine serovars. The antimicrobial susceptibility results revealed low-level resistance against 13 antimicrobial agents. Five strains exhibited resistance to nalidixic acid and intermediate resistance to ciprofloxacin with chromosomal (double) mutation at gyrA and parC while four strains possessed single mutation in parC. Salmonella Kentucky showed double mutation each at gyrA and parC. WGS analysis, revealed eight diverse sequence types (STs), the most common STs were ST-321 and ST-19 (n = 4) exhibited by S. Muenster and S. Typhimurium, respectively. Single Nucleotide Polymorphism (SNP)-based phylogeny analysis showed the 16 isolates to be highly related and fell into 8 existing clusters at NCBI Pathogen Detection. Curtailing the spread of resistant strains will require the establishment of continuous surveillance program at the state and national levels in Nigeria. This study provides useful information for further studies on antimicrobial resistance mechanisms in foodborne Salmonella species.

Antibacterial efficacy of ethyl acetate fraction of Psidium guajava leaf aqueous extract on experimental Escherichia coli (O78) infection in chickens.

AIM: This study was desingned to examine the efficacy of ethyl acetate fraction of aqueous extracted Psidium guajava leaves on chicks experimentally-infected with diarrheagenic strain of Escherichia coli O78. METHODS: A total of 60 ISA brown male chicks were randomly divided into 6 Groups of ten chicks each in separate cages. Group A was not infected and not treated. Groups B, C and D were infected and treated with extracts at a dose of 25, 50 and 100 mg/kg respectively for 10 days. Group E was infected and treated with oxytetracycline while Group F was infected, but left untreated. Chicks from all groups were closely monitored for clinical signs, body weight change and fecal bacterial shedding load during the course of the experiment. RESULTS: Diarrhea, vents pasted with feces, drop in feed intake accompanied by slow weight gain and decreased activity was observed in infected untreated groups. Groups treated with graded doses of the extract experienced a dose-dependent decreased in severity of the clinical signs shown compared to the infected untreated group. Bacterial shedding load was found to be lower in groups treated with the extract and oxytetracycline than those without intervention. CONCLUSION: Ethyl acetate soluble fraction of leaf extract of Psidium guajava effectively controlled diarrhea and decreased the severity of other clinical signs caused by experimental E. coli infections in chicks.

The first attempt of an active integrated laboratory-based Salmonella surveillance programme in the north-eastern region of Nigeria.

AIM: To identify the sources of Salmonella contamination, distribution, prevalence and antimicrobial susceptibility patterns, which have significant impact on public and animal health, and international trade. METHODS AND RESULTS: A total of 1888 samples were collected by stratified random sampling from 2009 to 2011 from cattle, camels, poultry, fish, vegetables and humans. All identified Salmonella isolates were serotyped and tested for antimicrobial susceptibility by MIC determinations. A total of 149 Salmonella isolates comprising 17 different serovars were obtained (7·9% prevalence). Salmonella Hadar (37%), S. Eko (17%), S. Enteritidis (10%), S. Kentucky (7%) and S. Uganda (7%) were isolated from different sources. The occurrence of antimicrobial resistance was generally low, but S. Enteritidis and S. Eko showed variable antimicrobial resistance patterns, while all S. Kentucky isolates were resistant to seven of 17 tested antimicrobials, including ciprofloxacin and nalidixic acid. Three S. Hadar isolates revealed reduced susceptibility to ciprofloxacin and susceptibility to nalidixic acid and harboured the plasmid-mediated quinolone resistance gene qnrS1. CONCLUSIONS: Salmonella serovars Hadar, Enteritidis and the previously very rarely reported Eko were the major serovars associated with human infections, animal and environmental contamination in the north-eastern region of Nigeria. SIGNIFICANCE AND IMPACT OF THE STUDY: These serovars constitute a health risk to poultry, environment and human population in the region.

The African medicines regulatory harmonization initiative: rationale and benefits.

Medicines regulation remains an important but neglected component of promotion and protection of public health because it helps to ensure that patients have access to quality, safe, and efficacious medicines. Investing in the African Medicines Regulatory Harmonization (AMRH) initiative provides an opportunity for strengthening regulatory capacity, cost-effective use of the limited financial and human resources, attainment of the three health-related Millennium Development Goals (MDGs), and promotion of trade and socioeconomic development for African countries and regions.

Tissue responses of Clarias gariepinus (Catfish) to experimental Eimeria subepithelialis infection.

Coccidial infection is a common feature in most African fishes. As a result of the chronic nature of the disease in fish, mortality is gradual (Kim and Paperna, 1993a) and is often overlooked in most farms, with losses becoming evident only at the end of the production cycle. This study evaluates the tissue responses of Clarias gariepinus (Catfish) in an experimental infection with Eimeria subepithelialis using 200 laboratory-bred Juveniles. Four groups (A, B, C and D) of 50 Juveniles each were preconditioned for 2 weeks. Groups A and B were infected per os with 200 E. subepithelialis sporocysts per juvenile, while groups C and D served as uninfected controls. At the onset of demonstrable oocysts in liver and spleen tissues, groups B and C were treated for a week in Amprolium(R) dip (0.6 mg/L) at the rate of 1 h a day. Five fish from each group were culled at intervals of 3 days to study the tissue responses to infection. At gross pathological level, intestinal and testicular congestion, splenomegaly and hepatomegaly were the common features of the infection. Histological lesions in the infected and treated group (group B) were confined to the small intestine while extra-intestinal lesions were seen in the infected and untreated fish (group A). The presence of pathological lesions in tissues, following experimental infection of C. gariepinus with E. subepithelialis sporocysts, indicated that the parasite is pathogenic to the catfish. Contrary to coccidiosis of terrestrial animals, sporulation of oocysts in piscine coccidiosis appeared to occur endogenously within the host. The use of Amprolium in the control of piscine coccidiosis is effective only when used early in the course of the infection, prior to onset of clinical signs. Proper certification of brooder and replacement stock, improved plane of nutrition and adequate stocking density are recommended aquacultural practices that may minimize the incidence of visceral coccidiosis in cultured fish.

Hybridization and contemporary evolution in an introduced cichlid fish from Lake Malawi National Park.

Rapidly evolving systems offer the chance to observe genetic and phenotypic change in real time. We exploit a well-characterized introduction of cichlid fish into Lake Malawi National Park to document a short history of habitat colonization and the evolution of genes and colour pattern. In the early 1960s, a fish exporter introduced individuals of Cynotilapia afra to a single site (Mitande Point) of Thumbi West Island and, as late as 1983, the species was confined to this location. In 2001, C. afra had colonized the entire perimeter of Thumbi West. In July of that year, we sampled C. afra individuals from six sites around the island and scored variation in dorsal fin colour as well as allelic diversity at six microsatellite loci. We found that, in two decades, C. afra had diverged into genetically distinct, phenotypically different northern and southern populations. We observed a high proportion of hybrids between the introduced C. afra and the native Metriaclima zebra on the southern coast of Thumbi West, and speculate that hybridization is facilitated by low water clarity at these windward sites. The short history of C. afra at Thumbi West is a microcosm of contemporary evolutionary divergence and may provide the opportunity to study the process from start to finish in genetic detail.

Antibody responses of goldfish (Carassius auratus L.) to DNA-immunisation at different temperatures and feeding levels.

DNA vaccines which work in winter are needed for fish. At 18 weeks after intramuscular injection of a plasmid containing lacZ goldfish at 15 degrees C had five-fold more antibody to betagalactosidase than those at 25 degrees C or those at

The safety and longevity of DNA vaccines for fish.

A plasmid that contained the cytomegalovirus (CMV)-promoter-driven lacZ reporter gene (pCMV-lacZ) remained in the epaxial muscle of five of eight goldfish as covalently closed circles, the most functional form of plasmid, for at least 70 days at 22 degrees. It was not present in the gills or elsewhere by polymerase chain reaction and was not integrated. Its expressed protein, Escherichia coli beta-galactosidase (beta-gal), which was in the injected myofibres, was detected in all the fish at 4-21 days and in about half the fish from 28 days until the end of the experiment at 70 days. The numbers of cells that secreted antibody to beta-gal in the kidney peaked at 14 days. Serum antibody and proliferating kidney cells to beta-gal were in all fish from 14 days with a plateau of the responses from 21 days onwards. The plasmid did not induce autoimmune-like antibodies to itself or to single- or double-stranded salmon testis DNA. Plasmids can therefore induce long-term foreign protein expression whilst inducing humoral and cell-mediated immunity without autoimmunity or integration in goldfish.

Mammalian granulocyte-macrophage colony-stimulating factor and some CpG motifs have an effect on the immunogenicity of DNA and subunit vaccines in fish.

A eukaryotic plasmid DNA carrying the AACGTT CpG motif in its ampR gene is a 'danger' signal for mice and caused an increase in the specific antibody titres of fish and mice after immunization with beta-galactosidase (beta-gal). A second pUC-based plasmid, which is inactive in mice and contains the GACGTC CpG motif in its cytomegalovirus (CMV) promoter, had no effect on antibody responses to beta-gal in either fish or mice. A synthetic oligonucleotide, which contains the GACGTT motif, potentiated antibody responses to co-administered beta-gal protein in mice, but not in fish. This is early evidence that lower and higher vertebrates recognize different unmethylated CpG motifs as 'danger' signals. In addition, plasmid DNA expressing mouse granulocyte-macrophage colony-stimulating factor (GM-CSF) had a marked effect on cytotoxic T-cell-like activity in fish by reducing the average number of myofibres that expressed beta-gal, 28 days after co-injection with plasmid DNA expressing beta-gal. Although the mechanism by which the mouse GM-CSF exerted its biological effects in fish is unknown, this finding might have important implications for fish vaccination, particularly when cytotoxic T cells may play a critical role.

Recent studies on the enterotropic strain of avian infectious bronchitis virus.

Avian infectious bronchitis (AIB) is an economically important disease of chickens. Recent studies have revealed enterotropism by at least one strain of AIB virus with pathological lesions in parts of the gut. This review highlights the findings of the studies so far made on this enterotropic strain of AIB virus.

The effects of oestrogen and progesterone on re-excretion of infectious bronchitis virus strain G (correction of strainG) in SPF chickens.

The effects of oestrogen and progesterone on the re-excretion of IB virus strain G in chickens infected at day-old was evaluated in this study. Following infection of the chickens at day-old, the birds were swabbed regularly from the trachea and cloaca until no more virus was isolated from either site. Between 10 and 14 weeks of age oestrogen and progesterone were administered by intramuscular injection to infected or control chickens. An infected but non-hormone injected group was maintained. All the birds were monitored for virus re-excretion and/or increase in serum neutralising (SN) antibody levels during this period of treatment. Similar parameters, and the egg production of these birds were evaluated when they reached their natural sexual maturity. At the end of the experiment, the birds were sacrificed and their internal organs especially the reproductive organs were examined for any pathological lesions. The acute phase of the disease following infection at day-old was typical of IB virus infection in terms of clinical signs and virus excretions from trachea and cloaca [1]. The injected hormones failed to induce virus re-excretion as no virus was isolated during this period. There was no change in the level of antibody during this period either. However, when the hens attained their natural sexual maturity, IB virus was re-excreted by all the birds. Isolations was also more frequent from the cloaca than from the trachea. The SN antibody levels of individual showed no definite pattern to correspond with periods of re-excretion in the birds up to the end of the experiment. The control birds remained normal throughout the period of the study.

Early pathogenesis in chicks of infection with an enterotropic strain of infectious bronchitis virus.

One-day-old specific-pathogen-free chicks were inoculated intranasally and intraocularly with infectious bronchitis virus (strain G). At days 1, 3, 5, 7, 10, and 14 postinfection, three birds were euthanatized, and the virus contents of both enteric tissues and some non-enteric tissues were assayed. Immunofluorescence and histopathological studies were also conducted. Six of 30 chicks died of nephritis between days 5-10 postinfection. Gross kidney lesions were the major pathological abnormalities. Inflammation was observed histologically in trachea, kidney, and rectum. High virus titers were found at various times in trachea, kidney, and all enteric tissues except for the jejunum. Relatively high titers of virus were still detectable at day 14 postinfection in the kidney, proventriculus, cecal tonsil, ileum, rectum, and bursa of Fabricius. Immunofluorescence staining showed viral antigens in enterocytes at the tips of villi in the ileum and rectum, and in the bursa. Viral antigens were also demonstrated in the epithelial cells of the trachea and in kidney tubules.

The effects of three reproductive hormones and cortisone on the replication of avian infectious bronchitis virus in vitro.

The in-vitro effects of three reproductive hormones (progesterone, oestrogen testosterone), and cortisone on the replication of infectious bronchitis (IB) virus strain G were investigated over a period of 36 hours using tracheal organ cultures as the culture system. The non-toxic concentration of each hormone for the culture system was first determined. These were found to be 3 micrograms/ml for progesterone, testosterone, and cortisone, and 1 micrograms/ml for oestrogen. The results were based on the assay of the extracellular virus production from both hormone treated and untreated infected cultures at specific intervals up to and including 36 hours. While oestrogen, testosterone, and cortisone were found to enhance the replication of the virus, no significant effect was noticed following treatment with progesterone.

The isolation and characterisation of six avian infectious bronchitis viruses isolated in Morocco.

The first isolation and characterisation of infectious bronchitis (IB) viruses from poultry flocks in Morocco are reported. Five isolates designated D, E, F, H and M were related serologically to the Massachusetts serotype, while the sixth, isolate G, was found to be different from any previously reported serotype of IB virus. Neutralising antibodies to isolate G have been detected in sera collected from commercial flocks in Britain, although the virus has not been isolated. While all six isolates caused respiratory disease typical of IB in experimentally infected 3-week-old specified pathogen-free (SPF) chickens, isolate G was unusual in that it could be isolated from several parts of the alimentary tract for up to 21 days post inoculation, and from the duodenum up to 28 days. H120 vaccines protected chicks challenged with isolates E and F but not isolate G.