Evaluation of berries of Phytolacca dodecandra for growth inhibition of Histoplasma capsulatum var.farciminosum and treatment of cases of epizootic lymphangitis in Ethiopia

Objective:To evaluate the berries of Phytolacca dodecundra(P.dodecandra) for its effect on Histoplasma cupsulatum var.farciminosum(HCF) and for the treatment of cases of epizootic lymphangitis(ELi.Methods:Samples were collected from un-ruptured nodules of cases of EL at Debre Zeit and Akaki(central Ethiopia).Mycologieal culture and isolation of HCF were performed at the Akliln Lemma Institute of Pathobiology.Phytochemical screening was done for n-butanol extract of P.dodecandra to delect alkaloids,saponins,phenolic compounds and flavonoids.The minimum inhibitory concentrations(MICs) and minimum fungicidal concentrations(MFCs) ol aqueous and n-butanol extracts of P.dodecandra against FICF were determined by agar dilution assay.For the in vivo trial.5%simple ointment was prepared from n-butanol extract and applied topically to 24(twelve early and twelve moderate) cases of F.L.Results:Phytochemical screening showed that n-butanol extract ol P.dodecandra was positive lor alkaloids.saponins and phenolic compounds but negative for flavonoids.The MFCs of n-butanol and aqueous extracts of P.dodecandra were(0.039%-0.078%) and(0.625%-1.250%),respectively.The MFCs of n-butanol and aqueous extracts of P.dodecandra were(0.078%t-0.156%)and(1.250%-2.500%),respectively.The MIC and MFC of ketoconazole(positive control) was(1.200×10^(-5)%-2.500×10^(-5)%) and(5.000× 10^(-5)%-1.000×10^(-4)%),respectively while growth was observed on free medium(negative control).From the total of 24 treated cases of EL,14(58.3%) responded lo treatment;however,10(41.7%) did not respond to treatment.There was no significant difference in the degree of response to treatment between early and moderate cases(χ~2=0.086:P=0.408.Conclusions:It can be concluded that n-butanol extract of P.dodecandra demonstrates antifungal effects while the aqueous extract shows no antifungal activity.

Aeromonas veronii isolated from climbing perch(Anabas testudineus)suffering from epizootic ulcerative syndrome(EUS)

Motile Aeromonads are responsible for various diseases in fishes that cause significant economic losses in aquaculture.Aeromonas spp.are also frequently isolated from fish suffering from Epizootic Ulcerative Syndrome(EUS).The present study was conducted to isolate and identify A.veronii from Vietnam climbing perch(Anabas testudineus)suffering from EUS,evaluate the pathogenicity of the isolates,and find out antibiogram profile and susceptibility of the isolates to medicinal plant extracts.Vietnam climbing perch(A.testudineus)suffering from EUS was collected from fish farms located at Savar,Dhaka and bacteria were isolated from the external ulcerative lesion,and kidney and liver of the infected fish on nutrient agar plates.Preliminary phenotypic identification of the bacterial isolates was performed following morphological,physiological and biochemical characterization tests.The 16S rRNA gene sequencing was accomplished for further molecular identification of five selected isolates.To evaluate the pathogenicity of the isolates in vivo artificial infection challenge test was conducted through immersion method.Another in vivo pathogenicity test was done at three different temperatures(18◦C,28◦C,and 37◦C)to examine the effects of temperature on the pathogenicity of the isolates.Ten commercial antibiotics were used to find out the antibiogram profile of the isolates.Finally,susceptibility of the isolates was screened against 19 medicinal plants extracts.A total of 38 out of 46 isolates isolated from Vietnam climbing perch(A.testudineus)suffering from EUS were phenotypically identified as Aeromonas sp.Five randomly selected isolates were further confirmed as A.veronii by analyzing the 16S rRNA gene sequence homology.The pathogenicity of the isolates was found moderate to high(41%-80%mortality)in the artificially challenged fish.The highest and lowest mortality was obtained in the fish challenged with the isolates BFKA33 and BFKA12,respectively.The isolates were found resistant to several antibiotics,including amoxicillin,ampicillin,penicillin-G,and vancomycin.Nevertheless,extracts of three medicinal plant(Allium sativum,Syzygium aromaticum,and Tamarindus indica)demonstrated antibacterial activity to the isolates(BFKA12,BFKA15,BFKA18,BFKA29,and BFKA33).

A serologic investigation of epizootic hemorrhagic disease virus in China between 2014 and 2019

Epizootic hemorrhagic disease virus(EHDV)is a member of the genus Orbivirus,family Sedoreoviridae.It was firstly recognized in 1955 to cause a highly fatal disease of wild white-tailed deer in America.So far,EHDV was detected and isolated in many wild or domestic ruminants,and widely distributed all over the world.Although the domestic cattle and sheep infected by EHDV were usually asymptomatic or subclinical,several outbreaks of epizootic hemorrhagic disease(EHD)in deer and cattle had been reported.Many EHDV strains were isolated and sequenced in last two decades in China,which promoted a general serologic investigation of EHDV in China.In this study,18,122 sera were collected from asymptomatic or subclinical domestic ruminants(cattle,cow,yaks,sheep,goats,and deer)in 116 regions belonging to 15 provinces in China.All the sera were tested by EHDV CELISA,and the results were obtained by big data analysis.EHDV infections were detected in the 14 of 15provinces,and only Tibet(average altitude≥4000 m)which was the highest province in China was free of EHDV.The numbers of seropositive collections in both bovine and goat/sheep were in an inverse proportion to the latitude.However,the seropositive rates in bovine were ranged from 0%to 100%,while the seropositive rates in goat/sheep were no more than 50%.The results suggested that bovine was obviously more susceptive for EHDV infection than goat and sheep,therefore might be a major reservoir of EHDV in China.The prevalence of EHDV was consistent with the distribution of Culicoides which were known as the sole insect vectors of EHDV.In particular,the seropositive rates of EHDV were very high in the southern provinces,which required the enhanced surveillance in the future.

Resolving the taxonomy of emerging zoonotic pathogens in the Trichophyton benhamiae complex

Species of the Trichophyton benhamiae complex are predominantly zoophilic pathogens with a worldwide distribution.These pathogens have recently become important due to their epidemic spread in pets and pet owners.Considerable genetic and phenotypic variability has been revealed in these emerging pathogens,but the species limits and host spectra have not been clearly elucidated.In this study,we used an approach combining phylogenetic analysis based on four loci,population-genetic data,phenotypic and physiological analysis,mating type gene characterization and ecological data to resolve the taxonomy of these pathogens.This approach supported the inclusion of nine taxa in the complex,including three new species and one new variety.Trichophyton benhamiae var.luteum var.nov.(“yellow phenotype”strains)is currently a major cause of zoonotic tinea corporis and capitis in Europe(mostly transmitted from guinea pigs).The isolates of the“white phenotype”do not form a monophyletic group and are segregated into three taxa,T.benhamiae var.benhamiae(mostly North America;dogs),T.europaeum sp.nov.(mostly Europe;guinea pigs),and T.japonicum sp.nov.(predominant in East Asia but also found in Europe;rabbits and guinea pigs).The new species T.africanum sp.nov.is proposed for the“African”race of T.benhamiae.The introduction to new geographic areas and host jump followed by extinction of one mating type gene have played important roles in the evolution of these pathogens.Due to considerable phenotypic similarity of many dermatophytes and phenomena such as incomplete lineage sorting or occasional hybridization and introgression,we demonstrate the need to follow polyphasic approach in species delimitation.Neutrally evolving and noncoding DNA regions showed significantly higher discriminatory power compared to conventional protein-coding loci.Diagnostic options for species identification in practice based on molecular markers,phenotype and MALDI-TOF spectra are presented.A microsatellite typing scheme developed in this study is a powerful tool for the epidemiological surveillance of these emerging pathogens.