Ethnoveterinary medicinal plants and their utilization by the people of Soro District, Hadiya Zone, southern Ethiopia.

BACKGROUND: Ethnoveterinary studies are important to maintain the sustainability of livestock health and support people's livelihoods through the provision of food, maintaining livestock health, and other biological resources. This study was carried out in Soro District, southern Ethiopia, to identify, document and analyse plant species with ethnoveterinary uses along with the associated indigenous and local knowledge. MATERIALS AND METHODS: Informants were selected using purposive (key informants) and systematic random sampling (general informants) methods. Data on ethnoveterinary plants and their uses were collected through semi-structured interviews, guided field walks, 13 focus group discussions with five to seven members in each and participant observation. Informant consensus factor and fidelity level were computed to identify the most common livestock ailment categories and the best plant species with ethnoveterinary use, respectively. Preference ranking methods were used to identify the potentially effective ethnoveterinary medicinal plants for the most frequently reported livestock ailments. The use diversity of multipurpose plants with ethnoveterinary importance was analysed using the analytical methods of ethnobotany including priority ranking, comparisons and important indices. The T-test statistic was used to compare knowledge differences among different social groups. RESULTS: A total of 132 plant species in 120 genera and 61 families were reported by informants as having ethnoveterinary uses. The plants are said to be used by the local communities in various ways to treat 50 livestock health problems. Higher number of informants (23.77%) cited Momordica foetida for the treatment of 16 livestock ailments. The highest informant consensus value for this species is associated with its use for treating blackleg in cattle; Nicotiana tabacum was cited for the treatment of 15 livestock ailments mainly recommended for the Lumpy Skin Disease/Ailment of bovines; Croton macrostachyus for treatment of 13 livestock ailments including wooden tongue, FMD in bovines; and Gymnanthemum amygdalinum for nine ailments mainly diarrhoea of all livestock types. Achyranthes aspera is claimed to provide the most effective treatment for Aspiration pneumonia (severe coughing in bovines, sheep and goats) alone, while Croton macrostachyus, Ximenia americana, Allium sativum and Juniperus procera were indicated as potential plant species to treat Lumpy Skin Disease in bovines in the order given. The fidelity level analysis showed that Datura stramonium, Dodonaea viscosa subsp. angustifolia and Asparagus africanus were potential medicinal plant species to treat the respective ailments of rabies, Peste des petits ruminants (PPR) and evil eye/spirit. Multipurpose plant species including Prunus africanus, Combretum molle and Afrocarpus falcatus have been highly threatened as indicated by direct matrix ranking mainly due to collection of fuel wood, construction materials and making household utensils, and farm implements rather than for other uses. CONCLUSION: Soro District has rich and diversified livestock herbal medicinal resources, and indigenous knowledge of remedy preparations and applications is transmitted through generation lines. This resource faces anthropogenic threats with deforestation being the leading factor. Consequently, ethnoveterinary medicinal plants continue to decline before adequate and proper scientific documentation and testing are made. There is a dire need for planning and implementation of appropriate in situ and ex situ conservation strategies and to strive towards ensuring the survival and sustainable utilization of such important plant resources of Soro District. This must be supported by further documentation of the associated indigenous knowledge and pharmacological testing of the key promising species including Balanites aegyptiaca (novel species/NS to treat specific ailment), Brugmansia suaveolens (novel species/NS reported first to treat Livestock ailments/LsAs), Euclea divinorum (NS to treat specific ailments), Grevillea robusta (NS), Hagenia abyssinica (NS for the reported specific ailment), Pentanema confertiflorum (NS), Juniperus procera (NS), Maesa lanceolata (NS), Millettia ferruginea (NS for reported specific ailments), Schrebera alata/NS, Securidaca longepedunculata, Spiniluma oxyacantha/NS, Vepris nobilis (novel species reported first to treat LsAs), Zanthoxylum asiaticum /NS and Ximenia americana (NS for specific ailments). This ethnoveterinary study attempted to fill part of the gaps concerning the prevalent livestock health problems and the associated indigenous and local knowledge in the area.

Development of a dual vaccine against East Coast fever and lumpy skin disease.

East Coast fever is an acute bovine disease caused by the apicomplexan parasite Theileria parva and is regarded as one of the most important tick-vectored diseases in Africa. The current vaccination procedure has many drawbacks, as it involves the use of live T. parva sporozoites. As a novel vaccination strategy, we have constructed the recombinant lumpy skin disease virus (LSDV) named LSDV-SODis-p67HA-BLV-Gag, encoding a modified form of the T. parva p67 surface antigen (p67HA), as well as the bovine leukemia virus (BLV) gag gene for the formation of virus-like particles (VLPs) to potentially enhance p67 immunogenicity. In place of the native sequence, the chimeric p67HA antigen has the human tissue plasminogen activator signal sequence and the influenza hemagglutinin A2 transmembrane domain and cytoplasmic tail. p67HA was detected on the surface of infected cells, and VLPs comprising BLV Gag and p67HA were produced. We also show that higher multiple bands observed in western blot analysis are due to glycosylation of p67. The two vaccines, pMExT-p67HA (DNA) and LSDV-SODis-p67HA-BLV-Gag, were tested for immunogenicity in mice. p67-binding antibodies were produced by vaccinated animals, with higher titers detected in mice vaccinated with the recombinant LSDV. This candidate dual vaccine warrants further testing in cattle.

Clinical Epidemiology, Pathology, and Molecular Investigation of Lumpy Skin Disease Outbreaks in Bangladesh during 2020-2021 Indicate the Re-Emergence of an Old African Strain.

Lumpy skin disease (LSD) emerged in Bangladesh in mid-2019, leading to great economic losses for cattle farmers. This study describes the recent occurrence of the LSDV in Bangladesh and examines the clinical manifestation of the disease in local cattle breeds, characteristic epidemiological features, and pathological findings in affected animals. In addition, a full-genome sequencing of two local LSDV isolates was carried out. A total of 565 animals from 88 households were investigated, and 165 samples (skin lesions, saliva, nasal discharge, feces, and milk) were collected for virus detection. Pathology and immunohistochemistry were performed on nodule biopsies. Fever, nodular skin lesions, and swelling of the joints were the most common clinical manifestations. Skin lesions had a higher concentration of viral DNA compared to other sample types and were therefore selected for virus isolation and characterization. Pathology of the LSD skin nodules comprised a granulomatous reaction in the dermis and hypodermis that extended to the surrounding tissues. Development of the skin lesions started with swelling of keratinocytes with cytoplasmic vacuolation, vasculitis, panniculitis, thrombosis, and infarction. Altogether, the LSDV produced transmural, hemorrhagic, necrotizing, proliferative and ulcerative dermatitis. The LSD viral antigen was detected occasionally in the macrophages, epithelial cells, and vascular smooth muscle cells. The complete genome sequence analysis revealed that the two Bangladeshi field strains (BD-V392.1 and BD-V395.1) were distinct from the contemporary field strains and were closely related to the ancestral African Neethling strain. The findings of this study will improve the diagnosis, monitoring, and control of LSD in Bangladesh.

Lumpy skin disease outbreak in cattle population of Chattogram, Bangladesh.

BACKGROUND: Lumpy skin disease (LSD) is an important viral disease causing significant economic losses in commercial livestock production. In mid-2019, an outbreak of LSD has been reported in cattle population from different parts of Bangladesh including Chattogram division. A cross-sectional surveillance study was undertaken from August 2019 to December 2019 to investigate the prevalence and associated risk factors of LSD in cattle in Chattogram district. METHODS: A total of 3,327 cattle from 19 commercial farms were examined for the LSD specific skin lesions and associated risk factors. A total of 120 skin biopsies were collected from the suspected animal for the confirmation of the disease using molecular detection and histopathological examination. Partial genome sequencing and phylogenetic analyses were performed on selected viral isolates. RESULTS: The overall clinical prevalence of LSD in the study population was 10% (95% confidence interval [CI]: 9.4%-11%) where the highest farm level outbreak frequency was 63.33% (95% CI: 45.51%-78.13%) and the lowest 4.22% (95% CI: 3.39%-5.25%). Crossbred and female cattle showed a significantly higher prevalence of the disease compared to their counterparts. Introduction of new animals in farms was found to be one of the most significant risk factors in the transmission of the disease. All suspected skin biopsies were positive for LSD virus (LSDV) infection with granulomatous and pyogranulomatous dermatitis was revealed on histopathology. Phylogenetic analysis based on the inverted terminal repeat region of the LSDV gene suggested that the locally circulating strain was closely related to the strains isolated from the Middle East and North African countries. CONCLUSIONS: The data generated in this study would be beneficial to the field veterinarians and animal health decision makers in the country as well as it will aid in taking appropriate measures to prevent further relapse or outbreak of this disease in future.

An Immunoperoxidase Monolayer Assay (IPMA) for the detection of lumpy skin disease antibodies.

During this study a new Immunoperoxidase Monolayer Assay (IPMA) was developed for the detection of antibodies against lumpy skin disease virus (LSDV) in an easy and low tech setting. Using two dilutions (1:50 and 1:300) in a duplicate format, the test was shown to be highly sensitive, specific and repeatable. In comparison to the VNT and a commercial ELISA, the LSDV-IPMA was able to detect the LSDV antibodies earlier in infected, vaccinated and vaccinated/infected animals. The assay is very flexible as it can be easily adapted for the detection of sheeppox or goatpox antibodies and it can be scaled-up to handle medium size sample sets by preparing the IPMA plates in advance. These plates are safe and can be handled in low biosafety level labs.

Avaliação pré-operatória por imagem da hidradenite supurativa / Preoperative imaging assessment of hidradenitis suppurativa

A hidradenite supurativa (HS) é uma doença inflamatória crônica da pele caracterizada por apresentar nodulações subcutâneas, dolorosas e com sinais flogísticos, inicialmente enrijecidas e que evoluem para consistência amolecida. Ocorre em 1 a 4% da população mundial. A sua etiologia ainda é pouco conhecida, sugere-se que aconteça devido à oclusão do ducto apócrino dos folículos pilosos por fatores precipitantes como fricção de tecido adiposo, higiene precária, entre outras. Seu diagnóstico é eminentemente clínico, pela identificação de lesões típicas recorrentes em forma de nodularidades, abcessos, tratos fistulosos ou cicatrizes. Não há testes patognomônicos. Sua evolução é variável e de difícil manejo, o qual pode ser feito com terapia tópica, sistêmica ou por exérese cirúrgica. Este trabalho revisa a avaliação por imagem da hidroadenite supurativa e demonstra imagens de um caso avaliado por ressonância magnética. A avaliação por exames de imagem, apesar de pouco específica para firmar diagnóstico, é muito útil na determinação da extensão da doença, assim como na exclusão de diagnósticos diferenciais, destacando-se o papel da ressonância magnética na avaliação das lesões anogenitais, com potencial de reduzir recorrências.

Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease characterized by painful deep-seated skin nodules with phlogistic signs, which are initially hard and progress to have a soft consistency. It occurs in 1­4% of the world population. Etiology of HS is still poorly understood and is suggested to occur due to occlusion of the apocrine duct of the hair follicles by triggering factors such as friction of the adipose tissue and poor hygiene, among others. Diagnosis is eminently clinical, through the identification of typical recurrent lesions that include nodules, abscesses, sinus tracts, or scars. There are no pathognomonic tests used to confirm its presence. Progression is variable and difficult to manage, which can be done with topical or systemic therapy or surgical excision. This work reviews the imaging assessment of HS and shows images of a case assessed by magnetic resonance imaging. Imaging assessment, although not specific enough for a diagnosis, is useful to determine the extent of the disease and to exclude differential diagnoses. Moreover, magnetic resonance imaging has an important role in the assessment of anogenital lesions and a potential to reduce recurrences.

Miofibromatose infantil: relato de uma rara doença / Infantile myofibromatosis: report of a rare disease

A miofibromatose infantil é uma rara doença que tem várias formas de apresentação. Habitualmente, manifesta-se com nódulos subcutâneos, que podem ou não estar associados à presença de nódulos viscerais. Deve-se estar apto a fazer o diagnóstico por meio do exame físico e de imagem, que evidenciarão o padrão das lesões para estadiar/classificar a doença. O tratamento ainda é controverso.

Infantile myofibromatosis is a rare disease with various presentations. Usually, such condition manifests itself as subcutaneous nodules, either in association or not with visceral nodules. The diagnosis should be achieved by means of physical examination and imaging studies, with emphasis on the lesions pattern to allow the staging and classification of the disease. The treatment is still controversial.

Evaluation of different diagnostic methods for diagnosis of Lumpy skin disease in cows.

Viral isolation, polymerase chain reaction (PCR), dot blot hybridization (DBH), and indirect enzyme-linked immunosorbent assay (iELISA) were used for the diagnosis of lumpy skin disease in clinically infected, fevered, and apparently normal dairy cows. Lumpy skin disease virus (LSDV) was isolated from skin biopsies and blood samples collected from clinically infected cows in percentages of 72% and 20%, respectively. The virus recovered from blood samples collected from fevered cows in percentage of 33.3%. Both PCR and DBH detected viral DNA in 100% of skin biopsies collected from clinically infected cows whereas the detection rates in blood samples collected from clinically infected animals were 100% and 84% using PCR and DBH, respectively. Viral DNA was detected in blood samples collected from fevered cows using PCR and DBH in percentages of 77.8% and 66.6%, respectively. Only 19.1% of blood samples collected from in-contact cows was positive for both of PCR and DBH. Detection rates of antibodies against LSDV using iELISA in serum samples collected from clinically infected and fevered cows were 56% and 11.1%, respectively, whereas all in-contact cows had no antibodies against the virus.

Evaluation of indirect fluorescent antibody test (IFAT) for the diagnosis and screening of lumpy skin disease using Bayesian method.

The performance of indirect fluorescence antibody test (IFAT) for serological diagnosis and screening of lumpy skin disease (LSD) was evaluated using methods without gold standard. Virus neutralization test (VNT) was used as the second test and the study sites were selected from two different geographical places in Ethiopia to get different disease prevalence. The analysis of conditional dependent Bayesian model for the accuracy of IFAT showed that sensitivity, specificity, prevalence of the population Pi(1) and the population Pi(2) were 0.92 (0.89-0.95), 0.88 (0.85-0.91), 0.28 (0.25-0.32) and 0.06 (0.048-0.075), respectively. The posterior inferences obtained for VNT sensitivity, specificity and conditional correlation between the tests for sensitivity (rhoD) and specificity (rhoDc) were 0.78 (0.74-0.83), 0.97 (0.95-0.99), 0.052 (-0.03-0.15) and 0.019 (-0.01-0.06), respectively. The interval estimation of conditional correlation for both sensitivity and specificity clusters around zero and thus conditional dependence between the two tests was not significant. Although accuracy measure would not be the only basis for test selection, the result of our study demonstrated that IFAT has a reasonable high accuracy to be used for the diagnosis and sero-surveillance analysis of LSD in the target population.

Elimination of toxicity and enhanced detection of lumpy skin disease virus on cell culture from experimentally infected bovine semen samples.

Lumpy skin disease virus (LSDV), a poxvirus of the genus Capripoxvirus, is shed in the semen of infected bulls. The screening of semen for infectious virus requires a sensitive diagnostic method. The isolation of the virus on cell cultures and/or the polymerase chain reaction (PCR) are sensitive diagnostic tests which may be used to screen semen for LSD viral DNA prior to artificial insemination. Although cell culture detects infectious virus and is a sensitive method, there are major difficulties in using this method due to the toxic effect of semen on the cells. The aim of this study was to find a method that decreases the toxic effect of semen and enhances the isolation of LSDV on cell culture. Semen samples from LSDV sero-negative bulls were collected and infected with a field isolate of LSDV, strain V248/93, with a titre of 6.5 log TCID50. The semen samples were treated with one of four different methods: centrifugation, serial dilution, filtration and chemical treatment with kaolin. The samples subjected to centrifugation, serial dilution and filtration were supplemented with gentamycin. Semen toxicity on cell cultures was eliminated when supernatants of semen samples centrifuged at 2000 rpm for 1, 3 and 5 min and serially diluted were used to inoculate confluent monolayer bovine dermis cells. The toxicity recorded when the pellet fractions of semen samples centrifuged for 5 min at 2000 rpm was comparable to results obtained from serially diluted samples supplemented with gentamycin. Filtration and kaolin treatment of semen samples did not remove the toxic effect.

The detection of lumpy skin disease virus in samples of experimentally infected cattle using different diagnostic techniques.

Lumpy skin disease (LSD) is a disease of cattle, primarily in Africa and Madagascar and rarely in the Middle East. It is caused by a capripoxvirus that belongs to the family Poxviridae. The disease is of economic importance in endemic areas. Effective control of LSD requires accurate and rapid laboratory techniques to confirm a tentative clinical diagnosis. Comparative studies on different diagnostic tests used at different stages of the disease have not been done. The aim of this study was to compare several of these tests. Six seronegative bulls, between 11 and 20 months of age, were infected intravenously and kept in an insect-free facility. The course of the infection was monitored. During a 3-month period blood samples and skin biopsies were collected for virus isolation and polymerase chain reaction (PCR). Skin biopsies were also examined using transmission electron microscopy (TEM). The incubation period in infected animals varied from 4-5 days. The length of the viraemic period did not correlate with the severity of clinical disease. Viraemia was detected from 1-12 days using virus isolation and from 4-11 days using the PCR, which is longer than has previously been reported. Virus was isolated from skin biopsies until Day 39 post infection (p.i.) and PCR could demonstrate viral DNA until Day 92 p.i. Transmission electron microscopy of negatively stained skin biopsies detected LSD virus only in one of the four bulls that developed skin lesions until Day 33 p.i. The PCR was a fast and sensitive method to demonstrate viral DNA in blood and skin samples. It could detect viral nucleic acid in skin lesions 53 days longer than virus isolation. Virus isolation from blood and skin samples was sensitive and reliable, but as a single test it may be too time-consuming to use although this depends on how rapidly the diagnosis must be confirmed. In conclusion, this study showed the PCR to be superior in detecting LSD virus from blood and skin samples. However, virus isolation is still required when the infectivity of the LSD virus is to be determined.

Use of a polymerase chain reaction assay to detect bovine herpesvirus type 2 DNA in skin lesions from cattle suspected to have pseudo-lumpy skin disease.

Beef cattle from a herd in north Alabama were examined because of an outbreak of nonfatal skin disease characterized by discrete circumscribed areas of inflammation that developed on the skin from the neck to the hips. Areas of inflammation, which tended to be superficial, underwent necrosis and scabbed over. The scabs eventually dropped off leaving discrete, round, whitish, hairless lesions that were 1.2 to 2.5 cm diameter. Because clinical signs were consistent with those expected with pseudo-lumpy skin disease (PLSD) caused by bovine herpesvirus type 2 (BHV-2), samples from 16 representative animals were submitted for BHV-2 testing. All 16 animals were seropositive for BHV-2, but the virus could not be isolated from skin biopsy specimens or buffy coat samples. Results of a polymerase chain reaction assay incorporating primers designed to amplify 2 DNA sequences from BHV-2 were positive for 3 of the 10 cattle, suggesting that skin lesions in these cattle were a result of PLSD. Our findings suggest that PLSD may be more common and widespread in the United States than suggested by the frequency with which BHV-2 has been isolated from cattle with PLSD-like skin lesions.

Trial of a capripoxvirus-rinderpest recombinant vaccine in African cattle.

Cattle were vaccinated with differing doses of an equal mixture of capripox-rinderpest recombinant viruses expressing either the fusion protein (F) or the haemagglutinin protein (H) of rinderpest virus. Animals vaccinated with 2 x 10(4) p.f.u. or greater of the combined viruses were completely protected against challenge, 1 month later, with both virulent rinderpest and lumpy skin disease viruses. Vaccination with any of the doses did not induce any adverse clinical response in the animals or transmission of the vaccine virus between animals. All cattle challenged 6 or 12 months after vaccination with 2 x 10(5) p.f.u. of the mixture of recombinant viruses were protected from severe rinderpest disease. Ten out of 18 were completely protected while the remaining 8 developed mild clinical signs of rinderpest. Cattle vaccinated with the recombinant vaccines after prior infection with the parental capripox virus showed more marked clinical signs of rinderpest after challenge with virulent rinderpest, but 9 out of 10 recovered, compared with 80% mortality in the unvaccinated controls.

Recombinant capripoxvirus expressing the hemagglutinin protein gene of rinderpest virus: protection of cattle against rinderpest and lumpy skin disease viruses.

A cDNA clone containing the complete coding sequence of the hemagglutinin (H) protein gene of the RBOK vaccine strain of rinderpest virus, under the control of the vaccinia late promoter p11, was inserted by homologous recombination into the thymidine kinase gene of the KS-1 strain of capripoxvirus. The recombinant virus produced authentic H protein as judged by its electrophoretic mobility, transport to the cell surface of infected lamb testis cells, and reactivity with monoclonal antibodies specific for the H protein of rinderpest virus. The recombinant virus induced significant levels of rinderpest virus neutralizing antibodies in vaccinated cattle and protected them from clinical rinderpest after challenge with a lethal dose of a highly virulent heterologous strain of the virus. Protection was achieved using vaccine doses lower than those used with a similar recombinant expressing the fusion protein gene of rinderpest. The parental KS-1 virus is widely used as a vaccine against capripox viruses and so the rinderpest recombinant acts as a dual vaccine to protect cattle against both rinderpest and lumpy skin disease.

Protection of cattle against rinderpest and lumpy skin disease with a recombinant capripoxvirus expressing the fusion protein gene of rinderpest virus.

Cattle were protected against challenge with rinderpest and lumpy skin disease viruses by vaccination with a recombinant capripoxvirus containing the fusion protein (F) gene of rinderpest virus. The minimum protective immunising doses for rinderpest and lumpy skin disease were 5.5 x 10(4) plaque forming units (pfu) and 1.5 x 10(3) pfu, respectively.

Single capripoxvirus recombinant vaccine for the protection of cattle against rinderpest and lumpy skin disease.

A recombinant capripoxvirus has been constructed containing a full-length cDNA of the fusion protein gene of rinderpest virus. The gene was inserted in the thymidine kinase gene of the capripox genome under the control of the vaccinia virus major late promoter p11 together with the Escherichia coli gpt gene in the opposite orientation under the control of the vaccinia early/late promoter p7.5. A vaccine prepared from this recombinant virus protected cattle against clinical rinderpest after a lethal challenge with a virulent virus isolate. In addition, the vaccine protected the cattle against lumpy skin disease.

A study of the pathology of lumpy skin disease in cattle.

Microscopic lesions in cattle infected with the virus of the Neethling form of lumpy skin disease comprised a granulomatous reaction in the dermis and hypodermis which extended to the surrounding tissue. During the early stages of the lesions a vasculitis and lymphangitis with concomitant thrombosis and infarction resulted in necrosis and oedema. A hallmark of the acute to subacute stages of the lesions was the presence of intracytoplasmic eosinophilic inclusions in various cell types. The inclusions consisted of the viroplasm which was identified as aggregates of electron-dense, finely granular to fibrillar deposits in which membrane-enclosed virions and occasional groups of tubular structures were observed. Various cytopathogenic changes were observed in cells exhibiting viral proliferation. The morphogenesis of the virions is discussed in relation to the cytopathogenic changes.