Salmonella Serovars, Antibiotic Resistance, and Virulence Factors Isolated from Intestinal Content of Slaughtered Chickens and Ready-to-Eat Chicken Gizzards in the Ilorin Metropolis, Kwara State, Nigeria.

Salmonellosis is one of the most common and widely distributed food-borne diseases, and the presence of antimicrobial-resistant Salmonella in poultry and poultry products is a global public health problem. Therefore, a cross-sectional study was conducted from November 2016 to July 2017 with an aim of determining the isolation rates of Salmonella species from the intestinal contents of slaughtered chickens, the most common serotypes that invade and colonize the tissues of chickens in Ilorin, and the susceptibilities of the isolated species to commonly used antibiotics. Four hundred samples of intestinal contents from apparently healthy slaughtered chickens and one hundred ready-to-eat chicken gizzards in Ilorin, Kwara State, were examined for the presence of Salmonella and their serotypes. Salmonellae were isolated and identified according to the techniques recommended by the World Health Organization: preenrichment, selective plating, biochemical testing, and serotyping. A total number of forty-three (43) Salmonella isolates consisting of 33 from intestinal contents and 10 from ready-to-eat chicken gizzards were isolated and identified. There was an overall Salmonella prevalence rate of 8.6% (43/500), and the isolates were distributed as follows: gizzard, 2% (n = 10) and intestinal contents, 6.6% (n = 33). The predominant serovars were Salmonella enterica subsp. enterica serovar 45: d: 1, 7 (16) and S. Haifa (5). All ready-to-eat chicken gizzards were associated with Salmonella enterica subsp. enterica serovar 45: d: 1, 7 (5). The Salmonella from intestinal contents belong to Salmonella enterica subsp. enterica serovars 45: d: 1, 7 (11) and S. Haifa (5). Salmonella species isolated were 100% resistant to ciprofloxacin, ampicillin, and ceftazidime. This is followed by cloxacillin (81%), tetracycline (75%), and sulfamethoxazole (67%). The Salmonella isolates were, however, 100% sensitive to enrofloxacin, 74% to streptomycin, and 72% to gentamycin antibiotics. The most common serotype was S. enterica subsp. enterica serovar 45: d: 1, 7. All the twenty five Salmonella serovars consisting of twenty-one serotypes (n = 21), two of the Salmonella that could not be cultured after enrichment, and the two that were contaminated with Proteus possessed the virulence genes of invA and stn. The Salmonella enterica subsp. enterica serovar 45: d: 1, 7 and S. Haifa possess virulence genes so they are potentially virulent for humans in this area. The national and local health authorities in Nigeria should improve hygiene measures especially at retail slaughter markets to reduce salmonellosis which is one of the most important food-borne diseases in humans.

Prevalence and distribution of dermatophytosis lesions on cattle in Plateau State, Nigeria.

BACKGROUND AND AIM: Dermatophytosis is an infection of the superficial, keratinized structures of the skin, nails, and hair of man and animals caused by a group of fungi called dermatophytes in the genera Trichophyton, Microsporum, and Epidermophyton. The prevalence of dermatophytosis among cattle in Nigeria and Plateau State, in particular, is yet to be fully determined. This study aimed to determine the prevalence and the distribution of dermatophytosis lesions on cattle in Plateau State, Nigeria. MATERIALS AND METHODS: Four hundred and thirty-seven cattle showing visible skin lesions suggestive of dermatophytosis were drawn from nine local government areas (three each) from the three senatorial districts of Plateau State, Nigeria. Skin scrapings were aseptically collected using a cross-sectional study, in which sampling units were selected using purposive sampling method. Samples were processed for both direct microscopic examination and isolation of dermatophytes in culture. The isolates were stained with lactophenol cotton blue and identified microscopically based on the size, shape, and arrangement of macro- and micro-conidia. The dermatophytes were further identified by determining the sequences of the internal transcribed spacer regions of their ribosomal DNA. Data were analyzed and presented as percentages, bar graph, and Chi-square test of association. p≤0.05 was considered statistically significant. RESULTS: The overall prevalence rate of bovine dermatophytosis in Plateau State was found to be 11.0%. Trichophyton verrucosum was more frequently isolated (54.2%) than Trichophyton mentagrophytes (45.8%). Age, breed, management practice, and season were significantly associated with the occurrence of the disease (p<0.05). CONCLUSION: Dermatophytosis among cattle may be of public health significance in Plateau State, Nigeria. This is the first report on the prevalence and distribution of dermatophytosis lesions on cattle from Plateau State, Nigeria.

Detection of NRAS G12D and NRAS G13C mutant genes among apparently healthy and haematologic malignant individuals in Federal Capital Territory, Nigeria.

Rat Sarcoma gene mutations is an important aspect in the management of hematologic malignancies globally. Unfortunately, this is not the trend in West Africa, including Nigeria. This study was aimed at detecting NRAS G12D and NRAS G13C mutant genes among apparently healthy and haematologic malignant individuals, and to explore their association with some clinical and demographic factors as well as disease status and progression. A total of 200 cfDNAs, 100 each from haematologic malignant patients and blood donors, respectively, were analyzed for the presence of NRAS gene mutations in codons 12 and 13. These mutations were tested using multiplex allele-specific PCR (AS-PCR). The mutations were detected by selective amplification using mutation-specific synthetic oligonucleotides. NRAS G12D and NRAS G13C mutations were 20.0% and 10.0%, respectively. In 17.5% of the 100 haemapoietic cancer patients, NRAS G12D mutant genes were seen while 7.5% of NRAS G13C mutation was found. Both mutant genes were observed in five healthy blood donors each. This result confirms the existence of NRAS mutations in Nigerian haemapoietic cancer patients and the preponderance of G-A transitions over G-T transversions. Mutant NRAS genes were associated with the types and stages of cancer, highlighting probable connection between mutation and increased susceptibility as well as quick progression of hematologic malignancies in the population studied. The result also highlighted higher risk of susceptibility/progression associated with leukemia than other hematopoietic cancers. We recommend more studies on NRAS mutation specifically targeted at improved diagnosis and prognostic therapy. The role of RAS mutation should be explored in other aside blood cancers in the Nigerian population.

Foot-and-mouth disease virus serotype SAT1 in cattle, Nigeria.

The knowledge of foot-and-mouth disease virus (FMDV) dynamics and epidemiology in Nigeria and the West Africa subregion is important to support local and regional control plans and international risk assessment. Foot-and-mouth disease virus serotype South African territories (SAT)1 was isolated, identified and characterized from an FMD outbreak in cattle in Nigeria in 2015, 35 years after the last report of FMDV SAT1 in West Africa. The VP1 coding sequence of the Nigerian 2015 SAT1 isolates diverges from reported SAT1 topotypes resulting in a separate topotype. The reporting of a novel FMDV SAT1 strain in the virus pool 5 (West and Central Africa) highlights the dynamic and complex nature of FMDV in this region of Africa. Sustained surveillance is needed to understand the origin, the extent and distribution of this novel SAT1 topotype in the region as well as to detect and monitor the occurrence of (re-)emerging FMDV strains.

Detection and Molecular Characterization of Foot and Mouth Disease Viruses from Outbreaks in Some States of Northern Nigeria 2013-2015.

Control measures for foot and mouth disease (FMD) in Nigeria have not been implemented due to the absence of locally produced vaccines and risk-based analysis resulting from insufficient data on the circulating FMD virus (FMDV) serotypes/strains. In 2013-2015, blood and epithelial samples were collected from reported FMD outbreaks in four states (Kaduna, Kwara, Plateau and Bauchi) in northern Nigeria. FMDV non-structural protein (NSP) seroprevalence for the outbreaks was estimated at 80% (72 of 90) and 70% (131 of 188) post-outbreak. Antibodies against FMDV serotypes O, A, SAT1, SAT2 and SAT3 were detected across the states using solid-phase competitive ELISA. FMDV genome was detected in 99% (73 of 74) of the samples from FMD-affected animals using rRT-PCR, and cytopathic effect was found in cell culture by 59% (44 of 74) of these samples. Three FMDV serotypes O, A and SAT2 were isolated and characterized. The phylogenetic assessments of the virus isolates showed that two topotypes of FMDV serotype O, East Africa-3 (EA-3) and West Africa (WA) topotypes were circulating, as well as FMDV strains belonging to the Africa genotype (G-IV) of serotype A and FMDV SAT2 topotype VII strains. While the serotype O (EA-3) strains from Nigeria were most closely related to a 1999 virus strain from Sudan, the WA strain in Nigeria shares genetic relationship with three 1988 viruses in Niger. The FMDV serotype A strains were closely related to a known virus from Cameroon, and the SAT2 strains were most closely related to virus subtypes in Libya. This study provides evidence of co-occurrence of FMDV serotypes and topotypes in West, Central, East and North Africa, and this has implication for control. The findings help filling the knowledge gap of FMDV dynamics in Nigeria and West Africa subregion to support local and regional development of vaccination-based control plans and international risk assessment.

Severe generalized skin lesions due to mixed infection with Sporothrix schenkii and Dermatophilus congolensis in a bull from Jos, Nigeria.

Sporothrix schenkii and Dermatophilus congolensis were isolated from a bull with severe generalized skin lesions. The lesions were thick, crusty and scabby on the dorsal part while few scabby and several nodular lesions were seen on the lower limbs especially the thighs. Scab samples and exudates from the nodules were aseptically collected and processed for bacteriology and mycology. Gram stained smears revealed Gram-positive, filamentous organism that had longitudinal and transverse septa suggestive of D. congolensis. Colonies on 5% defibrinated sheep blood agar were small, rough, grayish-white, ß-hemolytic and adherent to the medium. It was catalase positive, urease positive and fermented glucose and maltose but not sucrose, lactose, mannitol, sorbitol and xylose. Colonies on Sabouraud's dextrose agar were small, round, white and opaque, delicate and smooth. It liquefied gelatin and fermented glucose and sucrose but not galactose, menite, and glycerin. The isolate was Gram-positive, cigar-shaped and yeast-like suggestive of S. schenkii. Dermatophilosis is common in domesticated ruminants while sporotrichosis is very rare in cattle. This may be the first report of bovine sporotrichosis from Africa.

Epidemiological analysis, serological prevalence and genotypic analysis of foot-and-mouth disease in Nigeria 2008-2009.

The epidemiological situation of foot-and-mouth disease virus (FMDV) is uncertain in Nigeria, where the disease is endemic, and the majority of outbreaks are unreported. Control measures for FMD in Nigeria are not being implemented due to the absence of locally produced vaccines and an official ban on vaccine importation. This study summarizes the findings of a 3-year study aimed at quantifying the seroprevalence of FMD, its distribution in susceptible species and the genetic diversity of FMDV isolated from the Plateau State of Nigeria. A 29% FMD prevalence was estimated using 3ABC enzyme-linked immunosorbent assay (3ABC ELISA). Farms with suspected FMD nearby, with contact with wildlife, that used drugs or FMD vaccines or with >100 animals, and animals of large ruminant species and in pastures other than nomadic grazing were significantly (P < 0.05) associated with FMD. Antibodies against five FMDV serotypes, (A, O, SAT1, SAT2 and SAT3) were detected by the virus neutralization test (VNT) at various titres (<100->800) from all tested sera from most parts of the region. This is probably the first report of the presence of FMDV SAT3 in Nigeria. Further studies to investigate the potential probable presence and prevalence of SAT 3 virus in Nigeria are required. Tissue samples collected from clinical animals were positive for FMDV. Virus isolates were sequenced and confirmed as serotype A. All of the isolates showed marked genetic homogeneity with >99% genetic identity in the VP1 region and were most closely related to a previously described virus collected from Cameroon in 2000. This study provides knowledge on the epidemiological situation of FMD in Plateau State, Nigeria, and will probably help to develop effective control and preventive strategies for the disease in Nigeria and other countries in the West African subregion.

Characterization of field and vaccine infectious bursal disease viruses from Nigeria revealing possible virulence and regional markers in the VP2 minor hydrophilic peaks.

Outbreaks of infectious bursal disease in vaccinated chicken flocks are frequent in Nigeria. For the control of infectious bursal disease, live vaccines based on foreign infectious bursal disease virus (IBDV) strains are used. The present study investigated the phylogenetic relationship between field and vaccine IBDV strains from northwestern Nigeria. Thirty field IBDV strains and three commercial vaccines strains were characterized through sequencing the VP2 hypervariable region. In addition, the complete genome segment A coding region for two vaccines and two field strains was sequenced. The deduced amino acid sequences (position 212 to 331) of IBDV strains from Nigeria and other regions of the world were aligned and possible regional and virulence markers were identified associated with VP2 minor hydrophilic peaks. Reversion to virulence of a vaccine strain with a Q to L mutation at position 253 was observed. Phylogenetic analyses revealed a unique cluster of northwest Nigerian field IBDV strains alone or related to imported characterized classical and very virulent IBDV vaccines. The results suggest that when IBDV strains spread from their region of origin to a different region they mutate alongside indigenous field strains but may retain their identity on the VP2 region.

Serological evidence of brucellosis in goats in kaduna north senatorial district of kaduna state, Nigeria.

A cross-sectional study was carried out to determine the current status of Brucella antibodies in goats in Kaduna North Senatorial District of Kaduna State, Nigeria. A total of 442 serum samples (31 bucks and 411 does) were screened using Rose Bengal plate test (RBPT), serum agglutination test with ethylene diaminetetraacetic acid (SAT-EDTA), and lateral flow assay (LFA). Results. The prevalence of Brucella antibodies was found to be 25.8%, 11.1%, and 2.5% using RBPT, SAT-EDTA, and LFA, respectively. The prevalence in bucks was 32.3%, 3.2%, and 0.0% and 17.5%, 12.4%, and 3.9% in does using RBPT, SAT-EDTA, and LFA, respectively. The prevalence rates for goats less than one year of age using the tests were 1.5%, 0.0%, and 0.0%. While for those within the age bracket of one to three years, the rates were 19.4%, 10.5%, and 3.5%, respectively. The corresponding values for goats above 3 years of age were 34.2%, 15.2%, and 1.8%, respectively. The prevalence of brucellosis in goats in the study area is high which poses a threat to the development of the livestock industry and is of important zoonotic implications in Nigeria.

Antimycoplasmal activity of some plant species from northern Nigeria compared to the currently used therapeutic agent.

CONTEXT: Mycoplasma spp. are obligate parasites of humans and animals. But due to the special requirements needed to culture Mycoplasma in the laboratory, little or no research has been done to evaluate the efficacy of medicinal plants on the organism. OBJECTIVE: To screen medicinal plants traditionally used to treat infections for possible antimycoplasmal and cytotoxic activities. MATERIALS AND METHODS: Acetone extracts of 21 Nigerian medicinal plants were analyzed for antimycoplasmal and cytotoxicity activities using the metabolic inhibition and colorimetric methods, respectively. The extract with the best antimycoplasmal activities was also analyzed for its phytochemical constituents using the desktop method. RESULTS: Calotropis procera (Aiton) R.Br (Asclepiadaceae) extract had the best antimycoplasmal effect with a minimum inhibitory concentration (MIC) of 80 µg/mL and minimum mycoplasmacidal concentration (MMC) of 160 µg/mL. This extract contained saponins, tannins, cardiac glycosides, alkaloids, and flavonoids. The extract of Vernonia amygdalina Delile (Compositae) was the most cytotoxic with median lethal concentration (LC(50)) of approximately 17 µg/mL, and that of Anacardium occidentale L. (Anacardiaceae) was the least cytotoxic with an LC(50) of approximately 1919 µg/mL. DISCUSSION: Calotropis procera is a promising plant for an alternative antimycoplasmal agent because the crude acetone extract had a higher mycoplasmacidal activity than the conventional drug tylosin, which is currently used in treatment of the disease in Nigeria. CONCLUSION: The crude extract of Calotropis procera is worth investigating for the development of a potent agent against cattle Mycoplasma, which has long defied solution by conventional chemotherapy.

Frequency of involvement of pox virions in lesions of bovine dermatophilosis.

By combined electron microscopical and serological methods the rate of involvement of pox virions within lesions of bovine natural dermatophilosis was determined. Scabby lesions of 40 (77%) out of a total of 52 cases were positive for the virions, while sera of all infected animals which reacted positively for pox viral antibodies (LSD) was significantly higher (P less than 0.001) in comparison to those of healthy-appearing animals. The virions in all cases had a uniform oval to cylindrical morphology with a mean measurement of 110 X 280 nm. A synergistic interplay involving the initiation of the scabby lesions by the virions and their subsequent trapping within keratin deposits, followed by their release through the keratolytic activity of Dermatophilus congolensis appear to have obtained in cases.