Prevalence and Antimicrobial Resistance of Campylobacter coli and Campylobacter jejuni Isolated from Pig Guts, Pig Feces, and Surface Swabs from the Cutting Tables at Slaughterhouse and Taverns in Southern Benin.

Campylobacter food-borne infections are a serious public health problem. In Benin, there is a proliferation of pork consumption in several forms. This study aims to determine the prevalence and the antimicrobial resistance of Campylobacter coli and Campylobacter jejuni strains isolated from pig guts, pig feces, and surface swabs from the cutting tables in southern Benin. For this purpose, 200 samples of pig guts, 40 samples of swabs from the cutting table surface, and 8 samples of pig feces were collected and subjected to bacteriological examination. The method used for the identification of bacteria was microbiological culture combined with molecular identification by PCR. The identified strains were then subjected to antibiotic susceptibility testing according to the methodology recommended by the EUCAST. Antibiotic profiles were compared between strains isolated from pig guts, pig feces, and cutting table surfaces on the one hand and among the different sampling sites on the other hand. The results obtained show that 47.6% of the samples analyzed were contaminated by Campylobacter spp. Molecular identification revealed 34.7% of Campylobacter coli and 9.3% of Campylobacter jejuni. The study of antimicrobial susceptibility showed resistance to ciprofloxacin, 44% to ampicillin, 23.9% to erythromycin, 11% to gentamicin, and 10.1% to amoxicillin + clavulanic acid. In total, 90.8% of the isolated Campylobacter strains were multidrug resistant. The use of antimicrobials in livestock production systems has increased considerably, which could explain, at least partially, the prevalence of Campylobacter and the resistance of strains to antibiotics. To limit the risk of Campylobacter food-borne infections, it is therefore important to include Campylobacter in the list of pathogens to be tested during sanitary quality control of meat and meat products in Benin.

Whole genome analysis of rotavirus strains circulating in Benin before vaccine introduction, 2016-2018.

Species A rotaviruses (RVA) still play a major role in causing acute diarrhea in children under five years old worldwide. Currently, an 11-gene classification system is used to designate the full genotypic constellations of circulating strains. Viral proteins and non-structural proteins in the order VP7-VP4-VP6-VP1-VP2-VP3-NSP1-NSP2-NSP3-NSP4-NSP5/6 are represented by the genotypes Gx-P[x]-Ix-Rx-Cx-Mx-Ax-Nx-Tx-Ex-Hx, respectively. In Benin, ROTAVAC® vaccine was introduced into the Expanded Programme on Immunization in December 2019. To monitor circulating RVA strains for changes that may affect vaccine performance, in-depth analysis of strains prior to vaccine introduction are needed. Here we report, the whole-gene characterization (11 ORFs) for 72 randomly selected RVA strains of common and unusual genotypes collected in Benin from the 2016 to 2018 seasons. The sequenced strains were 15 G1P[8], 20 G2P[4], 5 G9P[8], 14 G12P[8], 9 G3P[6], 2 G1P[6], 3 G2P[6], 2 G9P[4], 1 G12P[6], and 1 G1G9P[8]/P[4]. The study strains exhibited two genetic constellations designed as Wa-like G1/G9/G12-P[6]/P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1 and DS-1-like G2/G3/G12-P[4]/P[6]-I2-R2-C2-M2-A2-N2-T2-E2-H2. Genotype G9P[4] strains possessed a DS-1-like genetic constellation with an E6 NSP4 gene, G9-P[4]-I2-R2-C2-M2-A2-N2-T2-E6-H2. The mixed genotype showed both Wa-like and DS-1-like profiles with a T6 NSP3 gene G1/G9P[8]/[4]-I1/I2-R1/R2-C1/C2-M1/M2-A1/A2-N1/N2-T1/T6-E1/E6-H1/H2. At the allelic level, the analysis of the Benin strains, reference strains (with known alleles), vaccine strains (with known alleles) identified 2-13 and 1-17 alleles for DS-1-like and Wa-like strains, respectively. Most of the study strains clustered into previously defined alleles, but we defined 3 new alleles for the VP7 (G3 = 1 new allele and G12 = 2 new alleles) and VP4 (P[4] = 1 new allele and P[6] = 2 new alleles) genes which formed the basis of the VP7 and VP4 gene clusters, respectively. For the remaining 9 genes, 0-6 new alleles were identified for both Wa-like and DS-1-like strains. This analysis of whole genome sequences of RVA strains circulating in Benin described genetic point mutations and reassortment events as well as novel alleles. Further detailed studies on these new alleles are needed and these data can also provide a baseline for studies on RVA in the post-vaccination period.

Toxicological Characterization of Ten Medicinal Plants of the Beninese Flora Used in the Traditional Treatment of Diarrheal Diseases.

The use of medicinal plants in traditional medicine is a common practice in developing countries. However, this unregulated or poorly rational use may present a dose-dependent risk of toxicity to humans. This study aimed to explore the phytochemical and toxicological characteristics of ten (10) plant species used in the traditional treatment of infectious diarrhea in Benin. The acute toxicity of aqueous and hydroethanolic extracts of Khaya senegalensis, Daniellia oliveri, Rauvolfia vomitoria, Vernonia amygdalina, Manihot esculenta, Ocimum gratissimum, Senna italica, Diospyros mespiliformis, Pterocarpus erinaceus, and Anacardium occidentale was evaluated following the OECD 423 protocol at a single dose of 2000 mg/kg. This safety test was complemented by a larval cytotoxicity test. Hematological and biochemical examinations, as well as a histological study of the liver and kidneys, were performed. Larval cytotoxicity was assessed by the sensitivity of Artemia salina larvae to different concentrations of the plant extracts studied. Testing for chemical compounds was performed on the basis of differential staining and precipitation reactions. The mean lethal concentration (LC50) was determined by the probit method. The qualitative phytochemical screening of the plants studied revealed the presence of catechic tannins, gallic tannins, flavonoids, anthocyanins and sterol-terpenes, alkaloids, saponosides, and reducing compounds. This composition varied according to the plants studied. Acute toxicity data indicated that there was no mortality and no structural and functional alterations of the liver and kidneys of treated animals. Larval cytotoxicity data suggest that the plants studied are not cytotoxic (LC50 ≥ 0.1 mg/mL). These observations reflect the safety of these plants and justify their use in traditional medicine in the treatment of many diseases including diarrheal diseases.

Effect of Rourea coccinea on ethanol-induced male infertility in Wistar albino rats.

Ethanol consumption is a risk factor of male infertility. The use of medicinal plants offers an alternative for the treatment of male infertility in developing countries. This study aimed to evaluate the Rourea coccinea effect on ethanol-induced male infertility in Wistar rats. Twenty-five (25) male Wistar rats were randomised into five groups of five rats and treated by oesophageal gavage over a 28-day period. Group 1 (negative control) received distilled water; Group 2 (positive control) received 30% ethanol at 7 mg/kg body weight; Group 3 (reference control) received 30% ethanol co-treated with the reference drug, clomiphene citrate; Groups 4 and 5 (test groups) received 30% ethanol co-treated with Rourea coccinea hydro-ethanolic extract at 200 and 400 mg/kg respectively. Testosterone hormone, sperm parameters and testicular histopathology were evaluated. Ethanol treatment induced a significant reduction (p < .05) in sperm count, motility, viability and a significant increase in sperm abnormalities because of the significant decrease (p < .05) in testosterone levels. These data correlate with the alterations observed in the seminiferous tubule on histopathological examination of the testes. However, co-treatment of ethanol with Rourea coccinea extract or the reference drug restored the ethanol-induced toxic effects on the reproductive organs, sperm profile and testosterone level.

Molecular characteristics of rotavirus genotypes circulating in the south of Benin, 2016-2018.

OBJECTIVE: Rotavirus remains the main causative agent of gastroenteritis in young children in countries that have not yet introduced the vaccine. In Benin, rotavirus vaccine was introduced late December 2019 into the EPI. This study aims to provide pre-vaccination era rotavirus genotyping data in Benin. These data can supplement data from the surveillance system of Ministry of Health of Benin which is supported by the World Health Organization (WHO). RESULTS: Of the 420 diarrheal stool samples, actively collected in southern Benin from July 2016 through November 2018 from children under 5 years old and suffering from gastroenteritis, 167 (39.8%) samples were rotavirus EIA positive. 186 (44.3%) samples contained amplifiable rotavirus RNA detected by qRT-PCR method and were genotyped using one-step RT-PCR multiplex genotyping method. G1P[8] represents the predominant genotype (32%) followed by the G2P[4] (26%), G3P[6] (16%), G12P[8] (13%) and mixed G and P types (1%). Four samples (2%) could not be assigned both G and P type specificity.

Tilapia (Oreochromis niloticus) as a Putative Reservoir Host for Survival and Transmission of Vibrio cholerae O1 Biotype El Tor in the Aquatic Environment.

Studies have reported the occurrence of Vibrio cholerae in fish but little is known about the interaction between fish and toxigenic V. cholerae as opposed to phytoplankton, which are well-established aquatic reservoirs for V. cholerae. The present study determined the role of tilapia (Oreochromis niloticus) as a reservoir host for survival and transmission of V. cholerae in aquatic environments. Three experiments were performed with one repetition each, where O. niloticus (∼2 g) kept in beakers were inoculated with four V. cholerae strains (5 × 107 cfu/mL). Firstly, infected tilapia were kept in stagnant water and fed live brine shrimp (Artemia salina) larvae daily. Secondly, infected tilapia were kept without feeding and water was changed every 24 h. Thirdly, infected tilapia were fed and water was renewed daily. Infected tilapia and non-infected controls were sacrificed on days 1, 2, 3, 7, and 14 post-inoculation and V. cholerae were enumerated in intestinal content and water. Another experiment assessed the transmission of V. cholerae from infected to non-infected tilapia. The study revealed that El Tor biotype V. cholerae O1 and V. cholerae non-O1 colonized tilapia intestines and persisted at stable concentrations during the second week of the experiment whereas the Classical biotype was undetectable after 1 week. In stagnant water with feeding, V. cholerae counts dropped to 105 cfu/ml in water and from 107 to 104 cfu/intestine in fish after 14 days. When water was renewed, counts in water decreased from 107 to 103 cfu/ml and intestinal counts went from 106 to 102 cfu/intestine regardless of feeding. All strains were transmitted from infected to naïve fish after 24 h of cohabitation. Tilapia like other fish may play an essential role in the survival and dissemination of V. cholerae O1 in aquatic environments, e.g., the seventh pandemic strains mostly. In this study, tilapia were exposed to high concentrations of V. cholerae to ensure initial uptake and follow-up studies with lower doses resembling natural concentrations of V. cholerae in the aquatic environment are needed to confirm our findings.

In vitro assessment of antibacterial activity from Lactobacillus spp. strains against virulent Salmonella species isolated from slaughter animals in Benin.

BACKGROUND AND AIM: Salmonella spp. are among the world's leading foodborne pathogens, found naturally in the intestines of many animals. Lactic acid bacteria, mainly Lactobacillus, are a promising alternative to antibiotics for animal and human health. This study aimed to assess the in vitro antibacterial activity of Lactobacillus spp. strains against virulent Salmonella spp. isolated from slaughter animals in Benin. MATERIALS AND METHODS: Eleven samples of raw cow's milk, five samples of breast milk, and six infant stool samples were taken. From these samples, strains of Lactobacillus were isolated and identified. The probiotic potential of each of the identified strains was characterized, and finally in vitro antibacterial activity of these strains was evaluated against three virulent strains of Salmonella spp. and a reference strain of Salmonella Typhimurium ATCC 14028. RESULTS: Out of the 22 samples collected, 20 strains of Lactobacillus spp. were isolated and identified. These strains included Lactobacillus plantarum (30%), Lactobacillus delbrueckii (25%), Lactobacillus casei (25%), Lactobacillus salivarius (15%), and Lactobacillus acidophilus (05%). Characterization of the probiotic potential of these strains showed that only 16 strains were resistant to pH=1.5. Fourteen of them were able to withstand the simulated gastric juice (pH 1.5+pepsin). The 14 probiotic strains showed very good antibacterial activity against virulent strains of Salmonella spp. with inhibition zone diameters ranging from 12.36±0.03 mm to 35.33±0.05 mm (R values>6 mm). CONCLUSION: From this study, Lactobacillus strains isolated from raw cow milk, breast milk, and infantile stool might be used as some valid candidates for probiotics. It also represents good alternatives for antibiotics in the fight against animal and human salmonellosis.

Epidemiological Survey of Rotaviruses Responsible for Infantile Diarrhea by the Immunomolecular Technique in Cotonou (Benin, West Africa).

Rotavirus remains the main causative agent of gastroenteritis in young children, in countries that have not yet introduced the vaccine. Benin, in order to implement the WHO recommendations, projects to introduce the rotavirus vaccine in 2018 as part of its Expanded Program on Immunization. But before the introduction of this vaccine, epidemiological data on rotavirus infections and rotavirus genotypes circulating in Benin should be available. The aim of this study is to generate epidemiological data on infantile rotavirus diarrhea in Benin. In order to determine the epidemiological characteristics and electrophoretypes of rotavirus responsible for gastroenteritis in diarrheic children aged 0 to 5 years, 186 stool samples were collected according to the WHO Rotavirus Laboratory Manual from March 2014 to February 2015 at Suru-Lere University Hospital Center. Detection of rotavirus antigen was performed by the ELISA test, followed by molecular characterization using polyacrylamide gel electrophoresis. 186 stool samples were analyzed for rotavirus, and seventy-three (39.2%) were found to be positive for rotavirus antigen by ELISA. Children aged 3 to 24 months were the most affected by rotavirus diarrhea in this study. Of the seventy-three children affected with rotavirus diarrhea, 27 (37%) had vomiting accompanied by dehydration and fever. Results based on electrophoresis showed that, among the 73 samples tested, 38 yielded typical rotavirus electrophoretic migration profiles.

Treatment of hypercholesterolemia: screening of Solanum macrocarpon Linn (Solanaceae) as a medicinal plant in Benin.

OBJECTIVE: Hypercholesterolemia is the greatest risk factor for cardiovascular diseases. The present study is conducted to evaluate the lipid lowering activity of leaves and fruits of Solanum macrocarpon, a vegetable, on Wistar rats experimentally rendered hypercholesterolemic by Triton X-100. MATERIALS AND METHODS: The leaves and fruits were administered (p.o.) for 7 days to rats at doses of 400 and 800 mg/kg of body weight. Atorvastatin was used as reference treatment drug. The data were analyzed by the Brown-Forsythe ANOVA, Dunnett's T3 multiple comparison test, and Dunnett's t test. All tests were done at the 5% significance level. RESULTS: Administration of S. macrocarpon (fruits as well as leaves) resulted in a statistically significant decrease in total cholesterol, LDL-cholesterol, VLDL-cholesterol, and triglycerides in the treated groups compared with the untreated hypercholesterolemic group, regardless of the administrated doses. A significant increase in HDL-cholesterol was observed in the treated groups. Hepatic disorders due to the Triton have been corrected by S. macrocarpon. CONCLUSIONS: This vegetable effectively suppresses experimental hypercholesterolemia in Wistar rats, suggesting a protective role in cardiovascular diseases. Its use by individuals at risk should be promoted.