In vivo evaluation of the antiviral activity of Cajanus cajan on measles virus.

Cajanus cajan, a tropical shrub, serves as source of food and traditional medicines. The evaluation of aqueous and ethanol extracts for activity against measles virus and toxicity to embryonated chicken eggs was carried out in this study. In vivo and in vitro assay techniques using embryonated chicken eggs and tissue culture (Hep-2 cell lines) as media for both virus cultivation and anti-virus assay showed that a hot-water extract yielded higher activity against measles virus. The hot-water extract of the stem yielded a Log(2) titre of 0.1 for the in vivo assay and an inhibition of cytopathic effect (CPE) in Hep-2 cells by 100% for the in vitro assay. At all concentrations of the extracts, there was a lowering of virus concentration (p = 0.05), indicated by hemagglutination (HA) titration, which is the advantage of HA titration over the tissue culture technique using CPE. This study validates embryonated chicken eggs as suitable media for anti-virus assay and the use of C. cajan in the treatment of some diseases of viral origin.

Evaluation of extracts of Anthocleista djalonensis, Nauclea latifolia and Uvaria afzalii for activity against bacterial isolates from cases of non-gonococcal urethritis.

Whole root preparations of three Nigerian medicinal plants, Anthocleista djalonensis, Nauclea latifolia and Uvaria afzalii, used traditionally in combination treatment of sexually transmitted diseases (STD), were extracted by maceration in ethanol, cold and hot water, respectively. The extracts were tested, by agar diffusion and macrobroth dilution methods, for activity against five strains of Staphylococcus aureus and two of Escherichia coli isolated from cases of STD and or urethritis. Four typed bacterial strains, S aureus ATCC 12600, Bacillus subtilis ATCC 6051, Pseudomonas aeruginosa ATCC 10145 and Escherichia coli ATCC 117755 were included as reference organisms. Ethanolic and cold-water extracts of Anthocliesta djalonensis exhibited activity against 9 and 7, respectively, of the 11 test organisms. They were bacteriostatic at minimum inhibitory concentrations (MIC) to the Gram positive strains but bactericidal to the Gram negative strains. Similar crude extracts of Uvaria afzalii showed bactericidal activity restricted to Gram positive (Staphylococcus aureus and Bacillus subtilis) strains. Nauclea latifolia extracts were bacteriostatic to both Gram positive and Gram negative strains. No test strain was susceptible to the hot water extracts of Nauclea latifolia but five and seven strains, were respectively susceptible to similar extracts of Anthocliesta djalonensis and Uvaria afzalii. Of the seven column chromatographic fractions of the ethanolic extract of Uvaria afzalii, F(ua-1) exhibited a bactericidal activity restricted to the Gram negative Escherichia coli strains, which were not susceptible to the crude extract. Fractions, F(ua-2), F(ua-3) and F(ua-4), like the crude extract, were bactericidal against the Gram positive strains only. Thus, partial purification seems to broaden the spectrum of activity and generally improve the potency of Uvaria afzalii. These results apparently justify the use of the three plants in treatment of STD.

Antibacterial activity of Harungana madagascariensis leaf extracts.

Aqueous extracts of the leaf of Harungana madagascariensis were analysed phytochemically and evaluated for antimicrobial activity against strains of Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Salmonella typhi and Pseudomonas aeruginosa. Glycosides, tannins, saponins, flavonoids and alkaloids were detected in the plant material. B. subtilis, E. coli and S. typhi, but not Ps. aeruginosa, showed susceptibility at MICs of 2.0 and 15.6 mg/mL; and MBCs of 2.0-3.9 mg/mL and 15.6-31.3 mg/mL, respectively, for the cold and hot extracts. Staph. aureus showed susceptibility only to the hot extract. Concentrations of 2.5-10.0 mg/mL of the cold extract killed over 7 log(10) of the test bacterial population within 30-60 min of exposure. The hot extract needed higher concentrations and longer treatment to achieve similar levels of bacterial cell killing. The results provide a rationalization for the traditional use of H. madagascariensis leaf extracts for the treatment of gastrointestinal disorders.

Evaluation of extracts of the root of Landolphia owerrience for antibacterial activity.

Ethanolic and aqueous (cold and hot) extracts of Landolphia owerrience root parts (whole-root, root-bark and root-wood) were tested for activity against ten bacterial strains using agar-well diffusion and macro-broth dilution methods, respectively. The ethanolic extracts of the whole-root and root-wood were active against 100 and 80% of the test organisms, respectively. Ethanolic and aqueous extracts of the root-bark were moderately active while the aqueous (cold and hot) extracts of the root-wood exhibited little or no activity. Out of the nine extracts prepared, 66.7% were active against Staphylococcus aureus ATCC 12600, 55.6% variously against each of Pseudomonas aeruginosa ATCC 10145 and local clinical isolates of P. aeruginosa, S. aureus, Escherichia coli and Salmonella typhi, 44.4% against Proteus sp., 33.3% against Bacillus subtilis ATCC 6051 and 22.2% against E. coli ATCC 11775. The agar-well-determined MIC values for the ethanolic whole-root extract (0.78-50 mg/ml) were higher (indicating lower activity) than the corresponding macro-broth-determined values (0.39-50 mg/ml) probably because of slow diffusion rates of the active constituents of the extract in agar. On the other hand, the differences could be due to the effects of DMSO used to dissolve the ethanolic extracts in the agar-well diffusion tests. Similar discrepancies in the MIC values detectable with the two test methods were apparent in the root-wood extract and the control drug, Gentamycin, except that in the latter the agar-well-determined MIC values (0.125-8.0 microg/ml) were lower than the macro-broth-determined values (0.125-64 microg/ml). The strong activity of the ethanolic extracts against known etiologic agents of diseases traditionally treated with L. owerrience root of similar preparations provides scientific justification for the use of the herb in ethnomedical practice in Nigeria.

Perceived causes and management of diarrhoea in young children by market women in Enugu State, Nigeria.

The aim of the study was to determine the perceptions of mothers regarding the causes and management of diarrhoea of their children aged 0-24 months. In Enugu State, Nigeria, 80 market women whose children had diarrhoea during last 6 months were interviewed fortnightly. When possible, the children were observed to determine the types of diarrhoea and treatments given. Fifty-three of the women brought their children to market, and 27 left their children at home. Seventy-one percent of the mothers perceived that diarrhoea was caused by teething. The most common types of diarrhoea occurring in these children were watery diarrhoea (59%) and the so-called teething diarrhoea (29%). Dysentery (6%) and jedi jedi or frothy and mucoid stools (4%) occurred less frequently. In 68% of the cases, drugs were used alone or in conjunction with salt-sugar solution (SSS) or other forms of treatment. These drugs were prescribed by medical personnel (40%), patent medicine dealers (23%), or mothers themselves (30%). About 26% and 39% of the mothers treated, respectively, watery and teething diarrhoeas with drugs only, while 23% used SSS alone. The drugs used were mainly antimicrobials (34%) and a combination of antimicrobial, antimalarial, antacid, analgesic, and some local herbal preparations (21%). The results of the study showed the evidence of unnecessary use of drugs and ignorance about their potential adverse effects. These underscore the need for appropriate primary care education among the market women in Nigeria.

Bacteriological quality of weaning food and drinking water given to children of market women in Nigeria: implications for control of diarrhoea.

Bacteriological quality of weaning food and drinking water given to 2 groups of children aged < or = years was evaluated by estimating bacterial cell count. One group consisted of those taken to market and the other of those left at home in the care of older siblings or house-helps. Bacterial counts (geometric mean) ranged from 5.02 +/- 1.82 to 8.70 +/- 1.0 log10 cfu per g or mL of food, and from 1.15 +/- 1.67 to 6.53 +/- 0.81 log10 cfu per g or 100 mL of water. Analysis of variance showed no significant difference in counts between types of food and between meals (breakfast and lunch). Bacterial contamination increased significantly with storage time, and was, in all circumstances except the water samples, significantly higher in foods given to children left at home. Reheated leftover foods also had significantly higher bacterial load than the freshly-cooked food. Coliform count varied significantly with source of drinking water. Poor hygiene standard (inferred from bacterial contamination) was generally observed among mothers weaning < or = 2-year-old children, while they were engaged in trading activities in the market, thus exposing their children to high risk of diarrhoea. Hygiene was significantly poorer in weaning of children left at home in the care of older siblings or house-helps. This implies that, in spite of their trading activities in the market, mothers still take better care of their babies than the older siblings or house-helps who may be inexperienced. These mothers may need education on childcare and food hygiene to suit to their trading activities, for example, during their monthly meetings. There is also a need to establish ORT (oral rehydration therapy) corners in the markets as part of the municipal services. This can be used not only for efficient and quick management of diarrhoea in the market but also for reinforcing hygiene education.

The effect of two child-care practices of market women on diarrhoea prevalence, feeding patterns and nutritional status of children aged 0-24 months.

A cross-sectional comparative study was conducted in seven markets in Enugu and Nsukka, Nigeria, to evaluate the child-care practices, occurrence of diarrhoea, feeding patterns and nutritional status among 506 and 157 children aged 0-24 months taken to the market (CTTM) by their mothers and those left at home (CLAH) respectively. A lack of a caretaker at home was the most common reason for taking children to the market. The majority (92%) of the caretakers at home were young (< 20 years). There was no difference in the occurrence of diarrhoea in the last month between the CTTM (39%) and the CLAH (41%) groups (p > 0.05). However, there were differences in reported diarrhoea episodes between children aged 0-6 months and higher age classes (chi 2 = 20.0; p = 0.003). Very few children (0.8%) were exclusively breastfed. More CTTM (58%) were still being breastfed than the 42% of CLAH (OR = 1.87; 95% C.I. = 1.27-2.37; p < 0.001). Children cared for at home had a slightly better but nonsignificant (p > 0.05) anthropometric status as characterised by weight-for-age and height-for-age. There was no relationship between feeding patterns and diarrhoea.

PIP: A cross-sectional comparative study was conducted in 7 markets in Enugu and Nsukka, Nigeria, to evaluate child care practices, the occurrence of diarrhea, feeding patterns, and nutritional status among 506 children aged 0-24 months taken to the market (CTTM) by their mothers and 157 similarly aged children left at home (CLAH). 49% of CTTM were infants, compared to only 35% of CLAH. Lack of a caretaker at home was the most common reason why mothers took their children to the market. 92% of caretakers at home were under 20 years old. 39% and 41% of children in the CTTM and CLAH groups, respectively, had diarrhea during the previous month. Differences were reported in the number of diarrhea episodes between children aged 0-6 months and older children. 0.8% of children were exclusively breast-fed, with 58% and 42% of children in the CTTM and CLAH groups, respectively, still being breast-fed. Children cared for at home had a slightly better but nonsignificant anthropometric status as characterized by weight-for-age and height-for-age. No relationship was observed between feeding patterns and diarrhea.

Recovery of velogenic Newcastle disease virus from dead and healthy free-roaming birds in Nigeria.

Fourteen Newcastle disease virus isolates were recovered, 10 from dead birds (nine chickens and one guinea fowl) and four from apparently healthy, free-roaming birds (one chicken, two ducks and one pigeon) in Nigeria. The pathogenicity indices showed all the isolates to be velogenic.

Incidence of rabies virus complement-fixing antibodies in unvaccinated dogs, humans and livestock in Anambra State of Nigeria.

A total of 376 serum samples from dogs, humans and livestock were examined for complement-fixing (CF) antibodies against rabies virus. High CF antibody titres (up to 1:1024) were detected among unvaccinated dogs aged 3 months and above as in vaccinated ones, thus establishing endemicity of the virus in the area. An antibody titre of 1:128 was detected in the serum of a puppy aged below 3 months and is unlikely to be due to residual maternal antibodies. It rather provides evidence for seroconversion at that age and further establishes endemicity of the virus. Moderate to high CF antibody titres (greater than or equal to 1:64) were detected in 44.9% of the unvaccinated dogs, 6.3% of the cattle and 2.8% of the humans. These antibody titres indicate either exposure to rabies virus or inapparent infection by rabies-related viruses.

Seroepidemiological evidence of widespread enteroviral infection in Anambra State, Nigeria.

Serum samples from 339 apparently healthy subjects, 298 adult blood donors and 41 infants aged 21 months and below, were screened for neutralising activity against poliovirus 1 and Coxsackieviruses A7 and B4. Among the adult serum samples 85% showed high activity (greater than or equal to 320) against poliovirus 1 while frequencies of similar activities were 65% and 64% for Coxsackieviruses A7 and B4 respectively. About 60% showed multiple activity against all three viruses; 15% against both Coxsackieviruses A7 and B4, and less than 5% for Coxsackievirus A7 and poliovirus 1, and Coxsackievirus B4 and poliovirus 1, respectively. Similar activities were detected in the sera of the infants. These show that enterovirus infections are endemic, multiple infections being the rule.

Bacterial agents associated with bronchopulmonary disorders in eastern Nigeria.

Altogether 16,539 sputum specimens were examined microbiologically from 1980 to 1984. Out of these 12,588 were screened by Ziehl-Neelsen's staining technique and 782 were (6.3%) found AFB-positive. Age and sex distributions of the AFB-positive individuals were statistically significant (at 0.05), incidence being most prevalent among those 20 years and above (90.2%) and among males (61.2%). From other specimens cultured, non-AFB organisms were isolated at the following frequencies: coliform-like organisms (15.1%), Streptococcus pneumoniae (55.5%). Klebsiella pneumoniae (5.3%), Streptococcus pyogenes (3.9%), Pseudomonas aeruginosa (3.4%), Haemophilus influenzae (3.0%). Proteus Spp. (0.7%) and Escherichia coli (0.5%). The antibiogram of these isolates revealed a high incidence of multiple antibiotic resistance, a situation that has most probably arisen from the high degree of antibiotic misuse in Nigeria.

Mutant identifying a third recombination group in a bunyavirus.

Only two recombination groups have been reported in genetic analyses of ts mutants of 10 different bunyaviruses from the Bunyamwera and California encephalitis serogroups, although three groups are expected from the tripartite structure of the genome of all members of the family Bunyaviridae. We describe now a ts mutant of Maguari virus, MAGts23(III), which recombined in both vertebrate (BHK-21) and invertebrate (Aedes albopictus) cells with mutants representing recombination groups I and II of this Bunyamwera serogroup virus. In addition, MAGts23(III) recombined with two mutants MAGts20 and MAGts21, provisionally identified as double mutants by their failure to recombine with group I or group II mutants, Mutant MAGts23(III) therefore represents a third bunyavirus recombination group. Mutant MAGts23(III) differed phenotypically from other bunyavirus mutants by growth restriction in BS-C-1 cells. Wild-type recombinants were obtained in the heterologous cross of MAGts23(III) and a group II mutant of Bunyamwera virus, but not in a cross with a group I mutant. The recombinants had the G protein of the Maguari virus parent and the N protein of the Bunyamwera virus parent. Analysis of the phenotypes of clones isolated at permissive temperature from the progeny of the other cross [MAGts23(III) and a group I mutant of Bunyamwera virus] indicated that recombination occurred in this cross, but that the possible recombinant phenotypes were not recovered with equal frequency. As a consequence, it has not been possible to obtain a gene assignment for group III from genetic data alone.

Genetic interactions among viruses of the Bunyamwera complex.

Seventy-seven temperature-sensitive (ts) mutants belonging to three antigenically distinct and geographically isolated members of the Bunyamwera complex--Batai virus, Bunyamwera virus, and Maguari virus--have been isolated after 5-fluorouracil treatment. High-frequency recombination was observed, and the mutants of each virus were classified into two groups, which were shown to be equivalent by heterologous recombination experiments. In most combinations heterologous recombination was less efficient than homologous recombination, but all crosses of group I and II mutants yielded viable recombinants. Recombination was an early event. Analysis by polyacrylamide gel electrophoresis of the proteins of the wild-type viruses and recombinant clones obtained from the six possible heterologous combinations of group I and II mutants indicated that recombination occurred by reassortment of genome subunits. Group I appeared to correspond to the genome subunit coding for the N protein, and group II corresponded to the G1/G2 determinant. The G1 (or G2 or both) protein was associated with neutralization specificity and plaque diameter, and the N protein was associated with plaque opacity. Complementation was observed between two nonrecombining mutants of Maguari virus belonging to group I, which may indicate that the N genome subunit codes for an additional protein. There appeared to be no genetic barrier to exchange of genetic material between Batai, Bunyamwera, and Maguari viruses in vitro, and it is concluded that the Bunyamwera complex is potentially a single gene pool if geographical and ecological constraints are discounted.