Seroprevalence and associated risk factors of contagious caprine pleuropneumonia in selected districts of South Wollo Zone Northeast Ethiopia.

Contagious caprine pleuropneumonia (CCPP) is a severe and devastating respiratory disease of goats, which is characterized by severe serofibrinous pleuropneumonia accompanied by high morbidity and mortality. A cross-sectional study was conducted from July 2022 to January 2023 to determine the seroprevalence of CCPP and identify risk factors associated with the occurrence of CCPP in goats in five selected districts of the South Wollo Zone of the Eastern Amhara region. A total of 384 sera samples were collected from goats and examined for antibodies specific to Mycoplasma capricolum subspecies capripneumoniae (Mccp) using Competitive Enzyme-Linked ImmunoSorbent Assay (cELISA) test. Out of the total examined sera, 26 samples were positive for CCPP, giving an overall seroprevalence of 6.7% (95% CI = 6.64-9.77). A seroprevalence of 5.05%, 4.65%, 2.78%, 12.90%, and 10.77% were recorded in Ambasel, Tehuledere, Kalu, Dessie Zuria and Kutaber districts, respectively. However, there was no statistically significant difference among these five districts (p > 0.05). The seroprevalence of CCPP varies significantly between age groups and agroecology (p < 0.05). However, the seroprevalence did not vary with sex, body condition score (BCS), and flock size (p > 0.05). Old-aged goats (OR = 4.10) and goats found in the lowlands (OR = 5.09) were at higher risk of infection with CCPP than young-aged goats and goats found in the highlands, respectively. In conclusion, the present seroprevalence investigation indicated the occurrence of CCPP in those selected study districts of the South Wollo Zone. Therefore, appropriate control measures, including avoiding the mixing of flocks and vaccination should be designed and implemented especially in the lowland areas and older goats to reduce the further spread and magnitude of the disease.

A molecular epidemiological investigation of contagious caprine pleuropneumonia in goats and captive Arabian sand gazelle (Gazella marica) in Oman.

BACKGROUND: Contagious caprine pleuropneumonia (CCPP) is a fatal WOAH-listed, respiratory disease in small ruminants with goats as primary hosts that is caused by Mycoplasma capricolum subspecies capripneumoniae (Mccp). Twelve CCPP outbreaks were investigated in 11 goat herds and a herd of captive Arabian sand gazelle (Gazella marica) in four Omani governorates by clinical pathological and molecular analysis to compare disease manifestation and Mccp genetic profiles in goats and wild ungulates. RESULTS: The CCPP forms in diseased and necropsied goats varied from peracute (5.8%), acute (79.2%) and chronic (4.5%) while all of the five necropsied gazelles showed the acute form based on the clinical picture, gross and histopathological evaluation. Colonies of Mccp were recovered from cultured pleural fluid, but not from lung tissue samples of one gazelle and nine goats and all the isolates were confirmed by Mccp-specific real time PCR. Whole genome-single nucleotide polymorphism (SNP) analysis was performed on the ten isolates sequenced in this study and twenty sequences retrieved from the Genbank database. The Mccp strains from Oman clustered all in phylogroup A together with strains from East Africa and one strain from Qatar. A low variability of around 125 SNPs was seen in the investigated Omani isolates from both goats and gazelles indicating mutual transmission of the pathogen between wildlife and goats. CONCLUSION: Recent outbreaks of CCPP in Northern Oman are caused by Mccp strains of the East African Phylogroup A which can infect goats and captive gazelles likewise. Therefore, wild and captive ungulates should be considered as reservoirs and included in CCPP surveillance measures.

Co-occurrence of contagious caprine pleuropneumonia and pasteurellosis in goats of sub-Saharan region: systematic review.

The reported incidences of co-participation of Mycoplasma capricolum capripneumoniae (Mccp) and Pasteurella multocida in increased severity and pathogenesis of goats with Contagious caprine pleuropneumonia (CCPP) in sub-Saharan Africa elicited the study's purpose. Using the preferred reporting items for systematic review and meta-analysis (PRISMA) guideline, two search engines, namely Google Scholar and PubMed, were queried to systematically review all the available literature on the current epidemiological status of CCPP and Pneumonic Pasteurellosis co-concurrently detected in goats and assess the available treatment and control measures and their challenges in the Sub-Saharan region. The search was limited to papers published between 1998 and 2024, whereby only peer-reviewed English articles were included in the study. Review papers, papers displaying abstracts only, duplicated information, papers beyond the sub-Saharan Africa region and papers published in other languages were excluded from the study. Only articles with full text and focused on goats were included for further screening process and review. A total of 3311 articles were retrieved from both databases, whereas only 58 articles met the inclusion criteria and hence were included in the data analysis. Only eight countries namely, Ethiopia, Kenya, Uganda, Nigeria, Sudan, Eritrea, Zambia and Tanzania reported the occurrence of CCPP and or Pasteurellosis: Ethiopia 23/58(39%), Tanzania 18/58 (31%), 1/58(2%) Nigeria, 1/58(2%) Zambia, 1/58(2%) Eritrea, Uganda 2/58 (3%), 2/58(3%) Sudan and Kenya 10/58(16%). Only 5/58 (9%) reported the occurrence of pneumonic pasteurellosis in Nigeria and Ethiopia. Only Tanzania (75%) and Ethiopia (33%) reported Mccp and Pasteurella multocida co-isolation and/ or detection in CCPP cases. Information on the antimicrobial susceptibility profile of Mccp and Pasteurella multocida from Sub-Saharan Africa was unavailable. One vaccine against CCPP, namely F-38 inactivated, and one vaccine against pneumonic pasteurellosis were identified to be developed and used in Sub-Saharan Africa. Developing bivalent candidate vaccines for both etiological agents is highly recommended.

The first identification of contagious caprine pleuropneumonia (CCPP) in sheep and goats in Egypt: molecular and pathological characterization.

Contagious caprine pleuropneumonia (CCPP) is one of the most fatal and contagious diseases of goats. To date, the occurrence of CCPP in Egypt has not been reported. During the period from 2017 to 2018, 200 goats and 400 sheep from Matrouh Governorate (Al Alamein and El Hammam cities) were suspected to have CCPP; animals were examined to confirm the presence of CCPP infection as well as the epidemiological status, clinical features, and molecular and histopathologic characteristics of lung tissues. Additionally, a treatment trial was performed to assess the efficacy of anti-mycoplasma therapy in the treatment of clinical cases of this disease. The occurrence of CCPP was 32.5% and 5% in goats and sheep, respectively, while case fatality was 30% and 8% in goats and sheep, respectively. The clinical forms of CCPP in both sheep and goats varied from per-acute to acute or chronic cases. Histopathological analysis of lung tissues from dead cases (either sheep or goats) revealed different stages of broncho- and pleuropneumonia ranging from per-acute to acute or chronic stages. Lung tissues showed severe congestion of interalveolar capillaries, flooding of alveoli and bronchi with a fibrinous exudate, a high degree of pleural thickening, and multifocal areas of necrosis that were sometimes sequestered in the fibrous capsule. Isolation of Mycoplasma capricolum subspecies capripneumoniae (Mccp) was confirmed in all dead cases by agar and broth culture methods and polymerase chain reaction. The treatment trial revealed that the marbofloxacin and spiramycin groups had a higher cure rate (70%) than the oxytetracycline group (40%) and a lower fatality rate (30%) than the oxytetracycline group (60%). Conclusively, infection with CCPP in goats and sheep is considered to be novel for Mccp in Egypt, where this species is considered to be the main pathogen in goats, not in sheep. Additionally, it could be concluded that treatment may be effective only if given early. Further comprehensive surveys are required to investigate the risk of CCPP in goats and sheep in all Egyptian governorates.

Evaluation of new generation macrolides for the treatment and metaphylaxis of contagious bovine pleuropneumonia (CBPP) in cattle experimentally infected with Mycoplasma mycoides subspecies mycoides.

BACKGROUND: Contagious bovine pleuropneumonia (CBPP) caused by Mycoplasma mycoides subspecies mycoides (Mmm) is an important disease of cattle that causes serious economic losses. With the known effectiveness of new generation macrolides, tulathromycin and gamithromycin were assessed in comparison with oxytetracycline as a positive control and saline as a negative control for effectiveness in inhibiting lung lesion development, promoting resolution, preventing spread and bacteriological clearance in susceptible local cattle breeds in two separate studies in Kenya and Zambia. Animals were monitored for clinical signs, sero-conversion as well as detailed post-mortem examination for CBPP lesions. RESULTS: Using the Hudson and Turner score for lesion type and size, tulathromycin protected 90%, gamithromycin 80%, and oxytetracycline 88% of treated animals in Kenya. In Zambia, all animals (100%) treated with macrolides were free of lung lesions, while oxytetracycline protected 77.5%. Using the mean adapted Hudson and Turner score, which includes clinical signs, post-mortem findings and serology, tulathromycin protected 82%, gamithromycin 56% and oxytetracycline 80% of the animals in Kenya whereas in Zambia, tulathromycin protected 98%, gamithromycin 94% and oxytetracycline 80%. The saline-treated groups had 93 and 92% lesions in Kenya and Zambia respectively, with Mmm recovered from 5/14 in Kenya and 10/13 animals in Zambia. Whereas the groups treated with macrolides were free from lesions in Zambia, in Kenya 5/15 tulathromycin-treated animals and 6/15 gamithromycin-treated animals showed lesions. Oxytetracycline-treated animals showed similarities with 3/14 and 4/15 showing lesions in Zambia and Kenya respectively and Mmm recovery from one animal in Kenya and six in Zambia. In both studies, lesion scores of saline-treated groups were significantly higher than those of the antibiotic treated groups (p < 0.001). In sentinel animals, CBPP lesions were detected and Mmm recovered from one and two animals mixed with the saline-treated groups in Kenya and Zambia respectively. CONCLUSIONS: This study demonstrated that tulathromycin, a mycoplasmacidal, can achieve metaphylactic protection of up to 80%, while non-recovery of Mmm from sentinels suggests macrolides effectiveness in preventing spread of Mmm. It is recommended that further studies are conducted to evaluate strategies comparing vaccination alone or combining vaccination and antibiotics to control or eradicate CBPP.

Improving Quality Control of Contagious Caprine Pleuropneumonia Vaccine with Tandem Mass Spectrometry.

Vaccines to protect livestock against contagious caprine pleuropneumonia (CCPP) consist of inactivated, adjuvanted antigens. Quality control of these vaccines is challenging as total protein quantification provides no indication of protein identity or purity, and culture is not an option. Here, a tandem mass spectrometry approach is used to identify the mycoplasma antigen contained in reference samples and in commercial CCPP vaccines. By the same approach, the relative amounts of mycoplasma antigen and residual proteins originating from the production medium are determined. Mass spectrometry allows easy and rapid identification of the peptides present in the vaccine samples. Alongside the most probable mycoplasma species effectively present in the vaccines, a very high proportion of peptides from medium constituents are detected in the commercial vaccines tested.

Respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection.

Contagious bovine pleuropneumonia (CBPP) is a severe disease caused by Mycoplasma mycoides subsp. mycoides (Mmm). Knowledge on CBPP pathogenesis is fragmented and hampered by the limited availability of laboratory animal and in vitro models of investigation. The purpose of the present study is to assess respiratory explants as useful tools to study the early stages of CBPP. Explants were obtained from trachea, bronchi and lungs of slaughtered cattle, tested negative for Mycoplasma spp. and for the major bacterial and viral respiratory pathogens. The interaction of Mmm with explant cells was studied by immunohistochemistry (IHC), double-labelling indirect immunofluorescence (DLIIF) and laser scanning confocal microscopy (LSCM). Mmm capability to survive and proliferate within the explants was evaluated by standard microbiological procedures. Finally, the putative cellular internalization of Mmm was further investigated by the gentamicin invasion assay. IHC and DLIIF indicated that Mmm can colonize explants, showing a marked tropism for lower airways. Specifically, Mmm was detected on/inside the bronchiolar and alveolar epithelial cells, the alveolar macrophages and the endothelial cells. The interaction between Mmm and explant cells was abolished by the pre-incubation of the pathogen with bovine anti-Mmm immune sera. Mmm was able to survive and proliferate in all tracheal, bronchial and lung explants, during the entire time course of the experiments. LSCM and gentamicin invasion assay both confirmed that Mmm can enter non-phagocytic host cells. Taken together, our data supports bovine respiratory explants as a promising tool to investigate CBPP, alternative to cattle experimental infection.

Search for OIE-listed ruminant mycoplasma diseases in Afghanistan.

BACKGROUND: Little is known about the occurrence of important diseases of ruminants in Afghanistan because of the conflict affecting the country over the last 40 years. To address this discrepancy, ruminant herds in Afghanistan were screened for OIE-listed mycoplasma diseases, contagious bovine (CBPP) and caprine pleuropneumonias (CCPP). RESULTS: Of the 825 samples from 24 provinces tested for serological evidence of CBPP caused by Mycoplasma mycoides subsp.mycoides, 20 (3.4%) had ELISA values greater than the positive threshold of 50% though all were less than 55%. Repeat testing of these suspect sera gave values below 50. A smaller number of sera (330) from cattle in nine provinces were also tested by the rapid latex agglutination test (LAT) for CBPP, 10 of which were considered suspect. However, no positive bands were seen when immunoblotting was carried out on all sera that gave suspect results. Serological evidence of Mycoplasma bovis was detected in half of 28 herds in eight provinces. The cause of CCPP, M. capricolum subsp. capripneumoniae was not detected in any of the 107 nasal swabs and lung tissue collected from goats in seven provinces though sample handling and storage were not optimal. However, strong serological evidence was detected in goat herds in several villages near Kabul some of which were over 50% seropositive by LAT and ELISAs for CCPP; immunoblotting confirmed positive results on a selection of these sera. CONCLUSIONS: The data presented here provide a first assessment of the occurrence of the two OIE listed mycoplasma diseases in Afghanistan. From the results of the testing bovine sera from the majority of provinces there is no evidence of the presence of CBPP in Afghanistan. However the samples tested represented only 0.03% of the cattle population so a larger survey is required to confirm these findings. Serological, but not bacterial, evidence was produced during this investigation to show that CCPP is highly likely to be present in parts of Afghanistan.

P19 contributes to Mycoplasma mycoides subsp. mycoides adhesion to EBL cells.

Mycoplasma mycoides subsp. mycoides (Mmm) is the causative agent of contagious bovine pleuropneumonia (CBPP). The virulent Mmm Ben-1 strain was isolated from the lung of a CBPP-infected cow in China in the 1950s. To attenuate the virulence of the Ben-1 strain and preserve its protective ability, the isolate was re-isolated after inoculation into the testicles of rabbits and into the rabbit thorax. As a result, after the subsequent isolates were continuously passaged 468 times in rabbits, its pathogenicity to cattle decreased. However, the molecular mechanisms leading to attenuation of the Mmm Ben-1 remain unknown. We compared the entire genomes of the Ben-1 strain and the 468 th generation strain passaged in rabbits (Ben-468) and discovered that a putative protein gene named p19 was absent from the Ben-468 strain. The p19 gene was cloned and expressed in Escherichia coli to obtain recombinant P19 (rP19). Western blot analysis demonstrated that the P19 protein is detected in the cell-membrane fraction, the cell-soluble cytosolic fraction and whole-cell lysate of the Mmm Ben-1 strain. The rP19 can interact with international standard serum against CBPP. Immunostaining visualised via confocal laser scanning microscopy indicated that P19 is able to adhere to embryonic bovine lung (EBL) cells, and this finding was also confirmed by a sandwich ELISA. We also found that anti-rP19 serum could inhibit the adhesion of the Mmm Ben-1 total proteins to EBL cells.

Morphological characterization and immunohistochemical detection of the proinflammatory cytokines IL-1ß, IL-17A, and TNF-α in lung lesions associated with contagious bovine pleuropneumonia.

Contagious bovine pleuropneumonia (CBPP), a severe respiratory disease, is characterized by massive inflammation of the lung especially during the acute clinical stage of infection. Tissue samples from cattle, experimentally infected with Mycoplasma mycoides subsp. mycoides Afadé, were subjected to histopathological and immunohistochemical examination in order to provide insight into innate immune pathways that shape inflammatory host responses. Lung lesions were characterized by vasculitis, necrosis, and increased presence of macrophages and neutrophils, relative to uninfected animals. The presence of three cytokines associated with innate inflammatory immune responses, namely, IL-1ß, IL-17A, and TNF-α, were qualitatively investigated in situ. Higher cytokine levels were detected in lung tissue samples from CBPP-affected cattle compared to samples derived from an uninfected control group. We therefore conclude that the cytokines TNF-α and IL-1ß, which are prevalent in the acute phase of infections, play a role in the inflammatory response seen in the lung tissue in CBPP. IL-17A gets released by activated macrophages and attracts granulocytes that modulate the acute phase of the CBPP lesions.

Free exopolysaccharide from Mycoplasma mycoides subsp. mycoides possesses anti-inflammatory properties.

In this study we explored the immunomodulatory properties of highly purified free galactan, the soluble exopolysaccharide secreted by Mycoplasma mycoides subsp. mycoides (Mmm). Galactan was shown to bind to TLR2 but not TLR4 using HEK293 reporter cells and to induce the production of the anti-inflammatory cytokine IL-10 in bovine macrophages, whereas low IL-12p40 and no TNF-α, both pro-inflammatory cytokines, were induced in these cells. In addition, pre-treatment of macrophages with galactan substantially reduced lipopolysaccharide (LPS)-induced production of pro-inflammatory cytokines TNF- and IL-12p40 while increasing LPS-induced secretion of immunosuppressive IL-10. Also, galactan did not activate naïve lymphocytes and induced only low production of the Th1 cytokine IFN-γ in Mmm-experienced lymphocytes. Finally, galactan triggered weak recall proliferation of CD4+ T lymphocytes from contagious bovine pleuropneumonia-infected animals despite having a positive effect on the expression of co-stimulatory molecules on macrophages. All together, these results suggest that galactan possesses anti-inflammatory properties and potentially provides Mmm with a mechanism to evade host innate and adaptive cell-mediated immune responses.

A large-scale genomic approach affords unprecedented resolution for the molecular epidemiology and evolutionary history of contagious caprine pleuropneumonia.

Contagious caprine pleuropneumonia (CCPP), caused by Mycoplasma capricolum subsp. capripneumoniae (Mccp), is a devastating disease of domestic goats and of some wild ungulate species. The disease is currently spreading in Africa and Asia and poses a serious threat to disease-free areas. A comprehensive view of the evolutionary history and dynamics of Mccp is essential to understand the epidemiology of CCPP. Yet, analysing the diversity of genetically monomorphic pathogens, such as Mccp, is complicated due to their low variability. In this study, the molecular epidemiology and evolution of CCPP was investigated using a large-scale genomic approach based on next-generation sequencing technologies, applied to a sample of strains representing the global distribution of this disease. A highly discriminatory multigene typing system was developed, allowing the differentiation of 24 haplotypes among 25 Mccp strains distributed in six genotyping groups, which showed some correlation with geographic origin. A Bayesian approach was used to infer the first robust phylogeny of the species and to date the principal events of its evolutionary history. The emergence of Mccp was estimated only at about 270 years ago, which explains the low genetic diversity of this species despite its high mutation rate, evaluated at 1.3 × 10(-6) substitutions per site per year. Finally, plausible scenarios were proposed to elucidate the evolution and dynamics of CCPP in Asia and Africa, though limited by the paucity of Mccp strains, particularly in Asia. This study shows how combining large-scale genomic data with spatial and temporal data makes it possible to obtain a comprehensive view of the epidemiology of CCPP, a precondition for the development of improved disease surveillance and control measures.

An international collaborative study to determine the prevalence of contagious caprine pleuropneumonia by monoclonal antibody-based cELISA.

BACKGROUND: Few serological tests are available for detecting antibodies against Mycoplasma capricolum subsp. capripneumoniae, the causal agent of contagious caprine pleuropneumonia (CCPP). The complement fixation test, the test prescribed for international trade purposes, uses a crude antigen that cross-reacts with all the other mycoplasma species of the "mycoides cluster" frequently infecting goat herds. The lack of a more specific test has been a real obstacle to the evaluation of the prevalence and economic impact of CCPP worldwide. A new competitive ELISA kit for CCPP, based on a previous blocking ELISA, was formatted at CIRAD and used to evaluate the prevalence of CCPP in some regions of Kenya, Ethiopia, Mauritius, Tajikistan and Pakistan in an international collaborative study. RESULTS: The strict specificity of the test was confirmed in CCPP-free goat herds exposed to other mycoplasma species of the "mycoides cluster". Prevalence studies were performed across the enzootic range of the disease in Africa and Asia. Seroprevalence was estimated at 14.6% in the Afar region of Ethiopia, whereas all the herds presented for CCPP vaccination in Kenya tested positive (individual seroprevalence varied from 6 to 90% within each herd). In Mauritius, where CCPP emerged in 2009, nine of 62 herds tested positive. In Central Asia, where the disease was confirmed only recently, no positive animals were detected in the Wakhan District of Afghanistan or across the border in neighboring areas of Tajikistan, whereas seroprevalence varied between 2.7% and 44.2% in the other districts investigated and in northern Pakistan. The test was also used to monitor seroconversion in vaccinated animals. CONCLUSIONS: This newly formatted CCPP cELISA kit has retained the high specificity of the original kit. It can therefore be used to evaluate the prevalence of CCPP in countries or regions without vaccination programs. It could also be used to monitor the efficacy of vaccination campaigns as high-quality vaccines induce high rates of seroconversion.

Interactions between Mycoplasma mycoides subsp. mycoides and bovine macrophages under physiological conditions.

We investigated the interactions of unopsonized and opsonized Mycoplasma mycoides subsp. mycoides (Mmm) with bovine macrophages in vitro. Mmm survived and proliferated extracellularly on bovine macrophage cell layers in the absence of Mmm-specific antisera. Bovine complement used at non-bactericidal concentrations did neither have opsonizing effect nor promoted intracellular survival, whereas Mmm-specific antisera substantially increased phagocytosis and Mmm killing. A phagocytosis-independent uptake of Mmm by macrophages occurred at a high multiplicity of infection, also found to induce the production of TNF, and both responses were unaffected by non-bactericidal doses of bovine complement. Bovine complement used at higher doses killed Mmm in cell-free cultures and completely abrogated TNF responses by macrophages. These results provide a framework to identify Mmm antigens involved in interactions with macrophages and targeted by potentially protective antibodies and point towards a pivotal role of complement in the control of inflammatory responses in contagious bovine pleuropneumonia.

Development of an improved vaccine for contagious bovine pleuropneumonia: an African perspective on challenges and proposed actions.

Contagious bovine pleuropneumonia (CBPP) caused by Mycoplasma mycoides subsp. mycoides (Mmm) is an economically very important cattle disease in sub-Saharan Africa. CBPP impacts animal health and poverty of livestock-dependent people through decreased animal productivity, reduced food supply, and the cost of control measures. CBPP is a barrier to trade in many African countries and this reduces the value of livestock and the income of many value chain stakeholders. The presence of CBPP also poses a constant threat to CBPP-free countries and creates costs in terms of the measures necessary to ensure the exclusion of disease. This opinion focuses on the biomedical research needed to foster the development of better control measures for CBPP. We suggest that different vaccine development approaches are followed in parallel. Basic immunology studies and systematic OMICs studies will be necessary in order to identify the protective arms of immunity and to shed more light on the pathogenicity mechanisms in CBPP. Moreover a robust challenge model and a close collaboration with African research units will be crucial to foster and implement a new vaccine for the progressive control of this cattle plague.

Isothermal loop-mediated amplification (LAMP) for diagnosis of contagious bovine pleuro-pneumonia.

BACKGROUND: Contagious Bovine Pleuropneumonia (CBPP) is the most important chronic pulmonary disease of cattle on the African continent causing severe economic losses. The disease, caused by infection with Mycoplasma mycoides subsp. mycoides is transmitted by animal contact and develops slowly into a chronic form preventing an early clinical diagnosis. Because available vaccines confer a low protection rate and short-lived immunity, the rapid diagnosis of infected animals combined with traditional curbing measures is seen as the best way to control the disease. While traditional labour-intensive bacteriological methods for the detection of M. mycoides subsp. mycoides have been replaced by molecular genetic techniques in the last two decades, these latter approaches require well-equipped laboratories and specialized personnel for the diagnosis. This is a handicap in areas where CBPP is endemic and early diagnosis is essential. RESULTS: We present a rapid, sensitive and specific diagnostic tool for M. mycoides subsp. mycoides detection based on isothermal loop-mediated amplification (LAMP) that is applicable to field conditions. The primer set developed is highly specific and sensitive enough to diagnose clinical cases without prior cultivation of the organism. The LAMP assay detects M. mycoides subsp. mycoides DNA directly from crude samples of pulmonary/pleural fluids and serum/plasma within an hour using a simple dilution protocol. A photometric detection of LAMP products allows the real-time visualisation of the amplification curve and the application of a melting curve/re-association analysis presents a means of quality assurance based on the predetermined strand-inherent temperature profile supporting the diagnosis. CONCLUSION: The CBPP LAMP developed in a robust kit format can be run on a battery-driven mobile device to rapidly detect M. mycoides subsp. mycoides infections from clinical or post mortem samples. The stringent innate quality control allows a conclusive on-site diagnosis of CBPP such as during farm or slaughter house inspections.

Antibody response to Mycoplasma capricolum subsp. capripneumoniae bacterium in small holder dairy goats in Tanzania.

Contagious caprine pleuropneumonia (CCPP), an Office International des Epizooties listed disease, can cause significant levels of morbidity and mortality in goats in Africa, the Middle East, and Western Asia. A cross-sectional study was conducted to determine the seroprevalence of CCPP, in two contrasting administrative districts of northern Tanzania, namely, Babati and Arumeru. A total of 337 serum samples were collected from January to July 2010, from apparently clinically healthy unvaccinated dairy goats breeds of different ages and sexes. Samples were subjected to monoclonal antibody-based competitive enzyme-linked immunosorbent assay for the specific measurement of antibodies to Mycoplasma capricolum subsp. capripneumoniae bacterium. The overall animal flock and village-level seroprevalence of CCPP was found to be 3.3 % [95 % confidence interval (CI) = 1.6-5.8], 9.6 % (95 % CI = 4.7-16.9), and 31.5 % (95 % CI = 15.6-56.5), respectively. There was no evidence (p = 0.659) of differences in seroprevalence between samples from the two administrative locations. Despite the fact that there was no significant difference (p > 0.05) in seropositivity between sex and age and between breeds sampled, there was a significant difference (p < 0.05) in seropositivity between the different physiological status groups examined with the non-lactating (dry) category of goats showing a higher seroprevalence. The findings of this survey revealed evidence of dairy goat exposure to M. capricolum subsp. capripneumoniae, although at a low prevalence. It is therefore advisable to include CCPP serology in the seromonitoring program so as to give a better indication of flock immunity. This should lead to the establishment of appropriate CCPP control measures in smallholder dairy goat flocks, which are increasingly being recognized for their value as a vital source of livelihood for resource poor livestock keepers in Tanzania.

Private sector involvement in the control of contagious bovine pleuropneumonia (CBPP) in the Kazungula district of Zambia benefitted the community and the control strategy.

Contagious bovine pleuropneumonia (CBPP) is a disease of economic importance that is widely distributed in sub-Saharan African and contributes significantly to cattle morbidity and mortality. Lack of resources to implement eradication measures has led to the disease becoming endemic in most areas in sub-Saharan Africa where governments have little resources and the majority of the people are poor. Usually, control and eradication of such diseases as CBPP is treated as a public good by governments and to achieve this, governments are usually assisted by nongovernment organisations, bilateral government programmes and international donors. The private sector, which usually is companies that run businesses to make profit, although not very well established in sub-Saharan Africa could play a big role in the eradication of CBPP in the region. This could play a dual role of promoting investment and also eradicate livestock diseases which have proved a menace in the livestock sector. This paper highlights the role played by the private sector in the control of CBPP in Zambia.

Complete genome sequences of Mycoplasma leachii strain PG50T and the pathogenic Mycoplasma mycoides subsp. mycoides small colony biotype strain Gladysdale.

Mycoplasma mycoides subsp. mycoides small colony biotype (SC) is the high-consequence animal pathogen causing contagious bovine pleuropneumonia. We report the complete genome sequences of the pathogenic strain M. mycoides subsp. mycoides SC Gladysdale and a close phylogenetic relative, Mycoplasma leachii PG50(T), another bovine pathogen of the M. mycoides phylogenetic clade.

Plasma levels of TNF-α, IFN-γ, IL-4 and IL-10 during a course of experimental contagious bovine pleuropneumonia.

BACKGROUND: Contagious Bovine Pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides, is widespread in sub-Saharan Africa. The current live vaccine T1/44 has limited efficacy and occasionally leads to severe side effects in the animals. A better understanding of the immune responses triggered by Mycoplasma mycoides subsp. mycoides and their role in disease progression will help to facilitate the design of a rational vaccine. Currently, knowledge of cytokines involved in immunity and immunopathology in CBPP is rather limited. The aim of this study was to characterize the in vivo plasma concentrations of the cytokines TNF-α, IFN-γ, IL-4, IL-10 and the overall role of CD4+ T cells in the development of cytokine levels during a primary infection. Plasma cytokine concentrations in two groups of cattle (CD4+ T cell-depleted and non-depleted cattle) experimentally infected with Mycoplasma mycoides subsp. mycoides were measured and their relationship to the clinical outcomes was investigated. RESULTS: Plasma cytokine concentrations varied between animals in each group. Depletion of CD4+ T cells did not induce significant changes in plasma levels of TNF-α, IL-4, and IL-10, suggesting a minor role of CD4+ T cells in regulation or production of the three cytokines during the time window of depletion (1-2 weeks post depletion). Unexpectedly, the IFN-γ concentrations were slightly, but statistically significantly higher in the depleted group (p < 0.05) between week three and four post infection. Three CD4+ T cell-depleted animals that experienced severe disease, had high levels of TNF-α and IFN-γ. Only one severely diseased non-depleted animal showed a high serum concentration of IL-4 post infection. CONCLUSIONS: Comparison of most severely diseased animals, which had to be euthanized prior to the expected date, versus less severe diseased animals, irrespective of the depletion status, suggested that high TNF-α levels are correlated with more severe pathology in concomitance with high IFN-γ levels.