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Foot-and-mouth disease (FMD) caused by FMD virus, is a highly infectious viral disease affecting livestock. Diagnosis plays a vital role in disease control and management during disease outbreaks. Available serotyping approaches are costly, time-consuming, requires complex equipment and trained personnel, warranting development of a diagnostic method that addresses this gap. In this retrospective study, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of serotype O FMDV was developed and evaluated using forty-three FMDV isolates from cattle. The specificity of the assay was tested against other FMDV isolates (serotypes A, SAT 1 and SAT 3). Only FMDV serotype O samples could be amplified within 26 min with an anneal derivative temperature of 87.02 degree celsius. Additionally, the assay amplified the viral protein 1 (VP1) gene with a detection limit of 0.0378 ng/μl. This molecular diagnostic approach has potential future application in improving FMDV surveillance as it provides baseline information for controlling FMD outbreaks in Tanzania and other Eastern and Southern African countries.

Plague has been a threat to human health in Tanzania since 1886. This zoonotic disease has established several endemic foci in the country, posing a risk of outbreaks. This study was conducted to investigate the presence ofBlood from 645 captured small mammals was screened for antibodies against the fraction 1 (F1) antigen ofSpecific antibodies againstThese results provide evidence of the circulation of

Infectious bursal disease (IBD) is a viral infection that affects young chicks. The IBD outbreaks in vaccinated chickens are reported in Tanzania frequently. The current study was conducted to find out the possible causes of vaccination failure focusing on knowledge and practices of vaccine sellers and users, the immunogenic potential of currently used vaccine and the phylogenetic relationship between the vaccine and the field strains. A cross-sectional study was performed to gather information on knowledge and practices from 384 poultry farmers and 20 veterinary outlets in Dar es Salaam. Results revealed inadequate knowledge of farmers in vaccine handling and administration and also breaches in the cold chain maintenance by vaccine sellers was apparent. A total of 60 chicks were experimentally vaccinated with Virgo 7 strain vaccine and titers of induced antibodies assessed. The vaccine induced adequate antibodies against IBDV, confirming its immunogenic efficacy. Isolated nucleic acids from the vaccine and field strains were sequenced and result shows that IBDV field isolates, are genetically different from the vaccine strains used in the country. The field isolates belong to the vvIBDV African types, while the vaccines belong to the vvIBDV European/Asian or classical virulent types. Putting together results from this study reveals multiple possible reasons which may contribute to vaccine failures. These include poor vaccine handling by farmers and vaccine sellers and the genetic disparity between the field and vaccine strains. It is therefore recommended that veterinary regulatory authorities should ensure good vaccine handling practices and considering local virus isolates during vaccine development. Key words: Infectious bursal disease (IBD), Infectious Bursal Disease Virus (vvIBDV), phylogenetic analysis, strains, vaccine, poultry farmers and antibodies.

Brucellosis is a zoonotic disease of importance to both public health and the livestock industry. The disease is likely to be endemic in Tanzania and little is reported on molecular characterization of Brucella species in pastoral settings. This study aimed at characterizing Brucella species (targeting genus Brucella) infecting humans, cattle and goat in Kagera region (Ngara and Karagwe districts) using real‐time PCR, PCR amplification of 16S rRNA genes and Sanger sequencing. Brucella spp. were detected in 47 samples (19 sera and 28 milk) out of 125 samples (77 sera, 35 milk and 13 aborted materials) using real‐time PCR. All aborted materials (13 samples) were negative to real‐time PCR. Out of the 47 real‐time PCR positive samples (28 milk and 19 sera), 20 samples (10 milk and 10 sera) showed an expected 16S rRNA gene PCR product. Sequence analysis and blasting confirmed the presence of Brucella spp. in pastoral areas of Kagera region. The Brucella spp. from Kagera were phylogenetically grouped in two clades and three branches all closer to B. melitensis, B. abortus and B. suis from USA, Sudan and Iran. However, they were distinct from other species isolated also in USA, New Zealand, Germany and Egypt. This was expected based on the distance between the geographical regions from which the data (nucleotides sequences from 16S gene sequencing) for the phylogeny reconstruction were obtained. This is the first study to report Brucella species identified using 16S rRNA gene sequencing in East and Central Africa. A livestock vaccination program re‐inforced with a high index of Brucella diagnosis is needed to eradicate brucellosis in animals and minimize suffering from Brucella infections in humans in Tanzania., Authors are reporting the presence of Brucella spp. in sera (from human) also in raw milk (from cattle and goat) which could raise the suspicions of the contribution of brucellosis to abortions observed in humans and animals interacting in pastoral areas of Kagera ecosystem, in Tanzania.

The control of brucellosis across sub-Saharan Africa is hampered by the lack of standardized testing and the use of tests with poor performance. This study evaluated the performance and costs of serological assays for human brucellosis in a pastoralist community in northern Tanzania. Serum collected from 218 febrile hospital patients was used to evaluate the performance of seven index tests, selected based on international recommendation or current use. We evaluated the Rose Bengal test (RBT) using two protocols, four commercial agglutination tests and a competitive enzyme-linked immunosorbent assay (cELISA). The sensitivity, specificity, positive predictive value, negative predictive value, Youden’s index, diagnostic accuracy, and per-sample cost of each index test were estimated. The diagnostic accuracy estimates ranged from 95.9 to 97.7% for the RBT, 55.0 to 72.0% for the commercial plate tests, and 89.4% for the cELISA. The per-sample cost range was $0.69–$0.79 for the RBT, $1.03–$1.14 for the commercial plate tests, and $2.51 for the cELISA. The widely used commercial plate tests performed poorly and cost more than the RBT. These findings provide evidence for the public health value of discontinuing the use of commercial agglutination tests for human brucellosis in Tanzania.

Small ruminants play an important role in the livelihoods of resource-constrained communities. This study was initiated because of a massive outbreak of a respiratory disease in sheep and goats in Loliondo area in Ngorongoro district of Arusha region in Tanzania in 2016. During flock examination, a total of 240 serum samples and 61 nasal swabs were collected. Antibodies to small ruminant morbillivirus, causative agent of peste des petits ruminants (PPR), were detected from sera using a competitive enzyme-linked immunosorbent assay. A multiplex reverse transcription real-time polymerase chain reaction assay was used to detect four pathogens: small ruminant morbillivirus, Mycoplasma capricolum subspecies capripneumoniae, Pasteurella multocida, and Capripoxvirus from the nasal swabs. Overall seroprevalence of PPR was 74.6%, with all four pathogens detected from nasal swabs. Co-infections of small ruminant morbillivirus and Mycoplasma capricolum subspecies capripneumoniae, small ruminant morbillivirus and Capripoxvirus, small ruminant morbillivirus and Pasteurella multocida, and Mycoplasma capricolum subspecies capripneumoniae and Capripoxvirus were also detected. Presence of PPR and the other diseases in this study provided insight into the severity of the outbreak in sheep and goats in Ngorongoro district. Thus, laboratory confirmation is critical for prompt and appropriate interventions to be made for control of diseases in sheep and goats with similar clinical signs. The findings also call for research into development of combined vaccines targeting common diseases of small ruminants in Tanzania.

Racheal A, Kasanga CJ, Byarugaba DK. 2018. Identification and characterization of Flavobacteriaceae from farmed Oreochromis niloticusand Clarius garieoinus. Bonorowo Wetlands 2: 42-50. Bacteria under family Flavobacteriaceae (in this study were also referred to as Flavobacteria) are important pathogens of fish, people, many other animals and plants. In this study, Flavobacteria from Nile tilapia (Oreochromis niloticus) and African catfish (Clarius gariepinus) were identified and characterized from the selected farms in Uganda. Gill and skin swabs were obtained from a total of 119 fish from 19 farms and were dissected aseptically to sample internal organs. The samples were inoculated onto Sheih media and incubated at 25°C for 48 hours. The suspected isolates were identified by colon characteristics, conventional biochemical tests and API 20 NE kits. The isolates were grouped into eight based on colon characteristic similarity. One isolate was selected per group for 16S rRNA gene sequencing and identified using the EZbiocloud.net ID software. Phylogenetic analysis of selected isolates was performed using the 16S rRNA gene sequences in BioEdit and MEGA 7.0.2 software. Basing on extrapolation of sequence analysis of the selected isolates, out of the 86 isolates, Myroides marinus was the most predominant species taking up 4 of the 8 groups (60 isolates) in 13 farms. The rest of the groups comprised of; Acinetobacter pitti, one group (6 isolates) in 6 farms, Chryseobacterium gambrini 2 groups (3 isolates) in 3 farms and one isolate was unidentified, in 3 farms. However, a total of 16 isolates did not grow on sub culturing. Phylogenetic analysis indicated that M. marinus isolates grouped with other M. marinus isolates from gene bank with significant intra-species diversity which was also observed with C. gambrini isolates. All the sampled farms had at least one isolate of a Flavobacterium from Tilapia and/or Catfish. Pathogenicity studies should be conducted on the isolates to establish their importance as fish pathogens and transmission dynamics so that an appropriate control measure can be recommended.

Summary Highly contagious transboundary animal diseases such as foot‐and‐mouth disease (FMD) are major threats to the productivity of farm animals. To limit the impact of outbreaks and to take efficient steps towards a timely control and eradication of the disease, rapid and reliable diagnostic systems are of utmost importance. Confirmatory diagnostic assays are typically performed by experienced operators in specialized laboratories, and access to this capability is often limited in the developing countries with the highest disease burden. Advances in molecular technologies allow implementation of modern and reliable techniques for quick and simple pathogen detection either in basic laboratories or even at the pen‐side. Here, we report on a study to evaluate a fully automated cartridge‐based real‐time RT‐PCR diagnostic system (Enigma MiniLab®) for the detection of FMD virus (FMDV). The modular system integrates both nucleic acid extraction and downstream real‐time RT‐PCR (rRT‐PCR). The analytical sensitivity of this assay was determined using serially diluted culture grown FMDV, and the performance of the assay was evaluated using a selected range of FMDV positive and negative clinical samples of bovine, porcine and ovine origin. The robustness of the assay was evaluated in an international inter‐laboratory proficiency test and by deployment into an African laboratory. It was demonstrated that the system is easy to use and can detect FMDV with high sensitivity and specificity, roughly on par with standard laboratory methods. This cartridge‐based automated real‐time RT‐PCR system for the detection of FMDV represents a reliable and easy to use diagnostic tool for the early and rapid disease detection of acutely infected animals even in remote areas. This type of system could be easily deployed for routine surveillance within endemic regions such as Africa or could alternatively be used in the developed world.

Summary Effective control and monitoring of foot‐and‐mouth disease (FMD) relies upon rapid and accurate disease confirmation. Currently, clinical samples are usually tested in reference laboratories using standardized assays recommended by The World Organisation for Animal Health (OIE). However, the requirements for prompt and serotype‐specific diagnosis during FMD outbreaks, and the need to establish robust laboratory testing capacity in FMD‐endemic countries have motivated the development of simple diagnostic platforms to support local decision‐making. Using a portable thermocycler, the T‐COR™ 8, this study describes the laboratory and field evaluation of a commercially available, lyophilized pan‐serotype‐specific real‐time RT‐PCR (rRT‐PCR) assay and a newly available FMD virus (FMDV) typing assay (East Africa‐specific for serotypes: O, A, Southern African Territories [SAT] 1 and 2). Analytical sensitivity, diagnostic sensitivity and specificity of the pan‐serotype‐specific lyophilized assay were comparable to that of an OIE‐recommended laboratory‐based rRT‐PCR (determined using a panel of 57 FMDV‐positive samples and six non‐FMDV vesicular disease samples for differential diagnosis). The FMDV‐typing assay was able to correctly identify the serotype of 33/36 FMDV‐positive samples (no cross‐reactivity between serotypes was evident). Furthermore, the assays were able to accurately detect and type FMDV RNA in multiple sample types, including epithelial tissue suspensions, serum, oesophageal–pharyngeal (OP) fluid and oral swabs, both with and without the use of nucleic acid extraction. When deployed in laboratory and field settings in Tanzania, Kenya and Ethiopia, both assays reliably detected and serotyped FMDV RNA in samples (n = 144) collected from pre‐clinical, clinical and clinically recovered cattle. These data support the use of field‐ready rRT‐PCR platforms in endemic settings for simple, highly sensitive and rapid detection and/or characterization of FMDV.

Rapid, reliable and accurate diagnostic methods provide essential support to programmes that monitor and control foot-and-mouth disease (FMD). While pan-specific molecular tests for FMD virus (FMDV) detection are well established and widely used in endemic and FMD-free countries, current serotyping methods mainly rely either on antigen detection ELISAs or nucleotide sequencing approaches. This report describes the development of a panel of serotype-specific real-time RT-PCR assays (rRT-PCR) tailored to detect FMDV lineages currently circulating in East Africa. These assays target sequences within the VP1-coding region that share high intra-lineage identity, but do not cross-react with FMD viruses from other serotypes that circulate in the region. These serotype-specific assays operate with the same thermal profile as the pan-diagnostic tests making it possible to run them in parallel to produce CT values comparable to the pan-diagnostic test detecting the 3D-coding region. These assays were evaluated alongside the established pan-specific molecular test using field samples and virus isolates collected from Tanzania, Kenya and Ethiopia that had been previously characterised by nucleotide sequencing. Samples (n = 71) representing serotype A (topotype AFRICA, lineage G-I), serotype O (topotypes EA-2 and EA-4), serotype SAT 1 (topotype I (NWZ)) and serotype SAT2 (topotype IV) were correctly identified with these rRT-PCR assays. Furthermore, FMDV RNA from samples that did not contain infectious virus could still be serotyped using these assays. These serotype-specific real-time RT-PCR assays can detect and characterise FMDVs currently circulating in East Africa and hence improve disease control in this region.

BACKGROUND: It has been demonstrated that infection with human parvovirus B19 (B19V) is associated with rash-fever illnesses. The present study aimed to investigate B19V as an aetiological agent of rash-fever syndromes in Congolese children confirmed as measles and rubella IgM-negative. An ELISA IgM test and PCR were performed to screen for B19V. METHODS: A total of 177 archived serum samples were randomly selected from the measles biobank of the National Institute for Biomedical Research (INRB). Samples were investigated for anti-B19V IgM and B19V DNA. These samples originated from children

SummaryAccurate, timely diagnosis is essential for the control, monitoring and eradicationof foot-and-mouth disease (FMD). Clinical samples from suspect cases are nor-mally tested at reference laboratories. However, transport of samples to these cen-tralized facilities can be a lengthy process that can impose delays on criticaldecision making. These concerns have motivated work to evaluate simple-to-usetechnologies, including molecular-based diagnostic platforms, that can bedeployed closer to suspect cases of FMD. In this context, FMD virus (FMDV)-specific reverse transcription loop-mediated isothermal amplification (RT-LAMP)and real-time RT-PCR (rRT-PCR) assays, compatible with simple sample prepa-ration methods and in situ visualization, have been developed which share equiva-lent analytical sensitivity with laboratory-based rRT-PCR. However, the lack ofrobust ‘ready-to-use kits’ that utilize stabilized reagents limits the deployment ofthese tests into field settings. To address this gap, this study describes the perfor-mance of lyophilized rRT-PCR and RT-LAMP assays to detect FMDV. Both ofthese assays are compatible with the use of fluorescence to monitor amplificationin real-time, and for the RT-LAMP assays end point detection could also beachieved using molecular lateral flow devices. Lyophilization of reagents did notadversely affect the performance of the assays. Importantly, when these assayswere deployed into challenging laboratory and field settings within East Africathey proved to be reliable in their ability to detect FMDV in a range of clinicalsamples from acutely infected as well as convalescent cattle. These data supportthe use of highly sensitive molecular assays into field settings for simple and rapiddetection of FMDV.IntroductionFoot-and-mouth disease (FMD) is a highly infectiousvesicular disease affecting both domesticated and wildcloven-hooved animals. Caused by FMD virus (FMDV),FMD affects over 100 countries worldwide, with diseasedistribution roughly reflecting economic development(Jamal and Belsham, 2013). Although the case-fatality rateof FMD is generally below 5%, the disease can beeconomically devastating: the annual global impact of FMD

A study was conducted to determine the serotypes of foot-and-mouth disease viruses (FMDV) circulating in African buffaloes (Syncerus caffer) from selected areas in Zambia. Sera and probang samples were collected between 2011 and 2012 and analysed for presence of antibodies against FMDV while probang samples were used to isolate the FMDV by observing cytopathic effect (CPE). Samples with CPE were further analysed using antigen ELISA. High FMD seroprevalence was observed and antibodies to all the three Southern African Territories (SAT) serotypes were detected in four study areas represented as follows: SAT2 was 72.7 percent; SAT1 was 62.6 percent; and SAT3 was 26.2 percent. Mixed infections accounted for 68.6 percent of those that were tested positive. For probang samples, CPE were observed in three of the samples, while the antigen ELISA results showed positivity and for SAT1 (n=1) and SAT2 (n=2). It is concluded that FMDV is highly prevalent in Zambian buffaloes which could play an important role in the epidemiology of the disease. Therefore livestock reared at interface with the game parks should be included in all routine FMDV vaccination programmes.

In this study, the seroprevalence of sparganosis and its relationship with sociodemographic factors in northern Tanzania have been assessed. A total of 216 serum samples from two rural districts, Monduli and Babati, were tested for sparganosis using an enzyme-linked immunosorbent assay. The seroprevalence of anti-sparganum IgG antibodies was 62.5% (95% confidence interval [CI] = 56.1–68.9) in all age groups. There were significant associations between district (relative risk [RR] = 1.95, 95% CI = 1.42–2.69), education (RR = 1.40, 95% CI = 1.15–1.70), and pet ownership with seropositivity (RR = 1.48, 95% CI = 1.02–2.16) based on univariate analysis. However, only the district was significantly associated with seropositivity (odds ratio = 4.20, 95% CI = 1.89–9.32) in binary logistic regression analysis. Providing health education to people residing in sparganosis-endemic areas is likely to improve the efficacy of preventative measures and reduce human disease burden.

The epidemiology of foot-and-mouth disease (FMD) in Africa is unique in the sense that six of the seven serotypes of FMD viruses (Southern African Territories [SAT] 1, SAT2, SAT3, A, O, and C), with the exception of Asia-1, have occurred in the last decade. Due to underreporting of FMD, the current strains circulating throughout sub-Saharan Africa are in many cases unknown. For SAT1, SAT2, and serotype A viruses, the genetic diversity is reflected in antigenic variation, and indications are that vaccine strains may be needed for each topotype. This has serious implications for control using vaccines and for choice of strains to include in regional antigen banks. The epidemiology is further complicated by the fact that SAT1, SAT2, and SAT3 viruses are maintained and spread by wildlife, persistently infecting African buffalo in particular. Although the precise mechanism of transmission of FMD from buffalo to cattle is not well understood, it is facilitated by direct contact between these two species. Once cattle are infected they may maintain SAT infections without the further involvement of buffalo. No single strategy for control of FMD in Africa is applicable. Decision on the most effective regional control strategy should focus on an ecosystem approach, identification of primary endemic areas, animal husbandry practices, climate, and animal movement. Within each ecosystem, human behavior could be integrated in disease control planning. Different regions in sub-Saharan Africa are at different developmental stages and are thus facing unique challenges and priorities in terms of veterinary disease control. Many science-based options targeting improved vaccinology, diagnostics, and other control measures have been described. This review therefore aims to emphasize, on one hand, the progress that has been achieved in the development of new technologies, including research towards improved tailored vaccines, appropriate vaccine strain selection, vaccine potency, and diagnostics, and how it relates to the conditions in Africa. On the other hand, we focus on the unique epidemiological, ecological, livestock farming and marketing, socioeconomic, and governance issues that constrain effective FMD control. Any such new technologies should have the availability of safe livestock products for trade as the ultimate goal.

Peste des petits ruminants (PPR) is an acute viral disease of small ruminants. The disease was first reported in Tanzania in 2008 when it was confined to the Northern Zone districts bordering Kenya. The present study was carried out to confirm the presence of PPR virus (PPRV) in Tanzania and to establish their phylogenetic relationships. Samples (oculonasal swabs, tissues and whole blood) were obtained from live goats with clinical presentation suggestive of PPR and goats that died naturally in Ngorongoro (Northern Tanzania) and Mvomero (Eastern Tanzania) districts. The clinical signs observed in goats suspected with PPR included fever, dullness, diarrhea, lacrimation, matting of eye lids, purulent oculonasal discharges, cutaneous nodules, erosions on the soft palate and gums and labored breathing. Post mortem findings included pneumonia, congestion of the intestines, and hemorrhages in lymph nodes associated with the respiratory and gastrointestinal systems. PPRV was detected in 21 out of 71 tested animals using primers targeting the nucleoprotein (N) gene. Phylogenetic analysis, based on the N gene, indicated that PPRV obtained from Northern and Eastern Tanzania clustered with PPRV strains of Lineage III, together with PPRV from Sudan and Ethiopia. The findings of this study indicate that there are active PPRV infections in Northern and Eastern Tanzania, suggesting risks for potential spread of PPR in the rest of Tanzania.

Aim: To investigate molecular and evolutionary characteristics of genes of fowlpox virus (FWPV) isolates from chickens in Tanzania. Study Design: Experimental. Place and Duration of Study: Faculty of Veterinary Medicine, Sokoine University of Agriculture, Morogoro, Tanzania; between November 2011 and October 2013. Methodology: Samples of cutaneous nodular lesions were collected from featherless parts of chickens (n = 154) suspected to have fowl pox in 14 regions of Tanzania followed by virus isolation, DNA extraction, polymerase chain reaction (PCR) amplification of the P4b gene, gel electrophoresis of PCR products, purification of PCR products, sequencing Original Research Article British Microbiology Research Journal, 4(7): 759-771, 2014 760 of purified PCR products and finally analysis of sequence data using standard procedures. Results: The disease was confirmed in 12 regions, out of 154 investigated samples 66 (42.86%) were found to contain FWPV, indicating that the 66 chickens from which the samples were collected had fowl pox as a result FWPV infection. Sequence analysis revealed that the Tanzanian FWPV isolates were 99.65 – 100% identical to each other and 99 – 100% identical to several published sequences of FWPV isolates from various countries in different continents of the world, including Europe and Asia. Phylogenetic analysis revealed that all Tanzanian isolates belong to clade A, subclade A1. Conclusion: Based on the findings of this study it is concluded that currently fowl pox is prevalent in several regions of Tanzania, caused by FWPVs which are genetically and phylogenetically closely related. However, these findings do not rule out the possibility of existence of genetic divergence among FWPVs currently prevalent in Tanzania. In order to rule out or detect genetic divergence (if any) among FWPVs currently prevalent in the country, other studies aimed at investigating molecular and evolutionary characteristics of genes in other genomic regions are highly recommended.

Foot and mouth disease is one of the world’s most important livestock diseases for trade. FMD infections are complex in nature and there are many epidemiological factors needing clarification. Key questions relate to the control challenges and economic impact of the disease for resource-poor FMD endemic countries like Zambia. A review of the control challenges and economic impact of FMD outbreaks in Zambia was made. Information was collected from peer-reviewed journals articles, conference proceedings, unpublished scientific reports, and personal communication with scientists and personal field experiences. The challenges of controlling FMD using mainly vaccination and movement control are discussed. Impacts include losses in income of over US$ 1.6 billion from exports of beef and sable antelopes and an annual cost of over US$ 2.7 million on preventive measures. Further impacts included unquantified losses in production and low investment in agriculture resulting in slow economic growth. FMD persistence may be a result of inadequate epidemiological understanding of the disease and ineffectiveness of the control measures that are being applied. The identified gaps may be considered in the annual appraisal of the FMD national control strategy in order to advance on the progressive control pathway.

Sero-surveillance was conducted to determine seroprevalence of peste des petits ruminant’s virus (PPRV) in sheep and goats population of Tanzania using samples collected in 2013 and 2015. A total of 3,838 samples were collected from villages in 14 of the 25 mainland regions. Samples were tested by competitive ELISA for detection of antibodies against PPRV. Overall, 998 of the samples were found to be positive for antibodies against PPR, giving a seroprevalence of 27.1%. In this study, there was no statistical significant difference of getting PPR between sheep and goats (odds ratio of 1.06, 95% CI 0.89-1.25). The overall seroprevalence indicates that PPR is prevalent in small ruminants in the study areas. The study also confirms the presence of antibodies against PPR in sheep and goats in regions of Tanzania that previously had little to no data on the disease, an indication that PPR is spreading within Tanzania with the possibility of spreading to neighboring countries.

Anthrax has become endemic throughout the upper Zambezi floodplain located in the Western Province of Zambia over the recent years. To date, no comprehensive study has been carried out to determine whether recurrence of anthrax outbreaks may be linked to differences in precipitation and human activities. Retrospective data for the period 1999 to 2007 showed that a total of 1,216 bovine cases of anthrax were reported. During the same period, 1,790 human anthrax cases and a corresponding case fatality rate of 4.63% (83/1,790) was documented in the upper Zambezi floodplain. Occurrence of human cases was highly correlated with cattle outbreaks (r = 0.94, p < 0.001). Differences in precipitation were significantly associated with the occurrence of anthrax outbreaks (χ2 = 4.75, p < 0.03), indicating that the likelihood of outbreaks occurring was higher during the dry months when human occupancy of the floodplain was greater compared to the flooding months when people and livestock moved out of this region. Human dependency on the floodplain was shown to significantly influence the epidemiology of anthrax in the upper Zambezi floodplain of western Zambia. Methods for mitigating anthrax outbreaks by disrupting the cycle of transmission are herein highlighted. ⓒ 2012 The Korean Society of Veterinary Science. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

A practical sampling method for bursal tissue using ordinary paper for molecular diagnosis of infectious bursal disease (IBD) was established. IBD virus-infected bursa was directly smeared on chromatography paper, filter paper, or stationery copy paper and was then fixed with absolute ethanol, Tris-HCl-saturated phenol, or phenol:chloroform:isoamyl alcohol (25:24:1). Flinders Technology Associates (FTA) card, which is designed for the collection of biological samples for molecular detection, was also used. After storage at 37 C for up to 30 days, total RNA directly extracted from the tissue fixed on the papers and FTA card were subjected to reverse transcriptase-polymerase chain reaction (RT-PCR) for detection of IBD virus (IBDV) RNA. In addition, the ability of each chemical used in the fixation and the FTA card to inactivate IBDV was evaluated. Regardless of the paper quality, storage period, and fixation method, IBDV RNA was consistently detected in all of the samples. IBDV in the bursal tissue was inactivated with phenol but not with ethanol or the unknown chemicals in FTA card. These results show that ordinary papers sustain the viral RNA, as does FTA card, but phenol fixation is superior to FTA card in inactivating IBDV. The new sampling method using ordinary paper with phenol fixation is safe, inexpensive, simple, and easy, and is thus suitable for conducting a global survey of IBD even where laboratory resources are limited. This practical method should contribute to the control of IBD worldwide.

To the Editor: African swine fever (ASF) is a highly contagious and deadly hemorrhagic disease of domestic pigs caused by African swine fever virus (ASFV), a double-strand DNA virus of the family Asfarviridae and genus Asfivirus (1). Twenty-two ASFV genotypes (I–XXII) have been identified on the basis of nucleotide sequencing of the variable 3′-end of the B646L gene encoding the major capsid protein p72 (2,3). Historically, all ASFV p72 genotypes have been circulating in eastern and southern Africa, and genotype I has been circulating in Europe, South America, the Caribbean, and western Africa (2,3). Spread of ASFV beyond traditional geographic boundaries occurred with incursion of p72 genotype II into the Republic of Georgia and its subsequent spread into Armenia, Azerbaijan, and Russia (4,5) and incursion of genotype IX into western Africa (6). ASFV circulating in Tanzania has p72 genotypes X, XV, and XVI (7–10). We describe incursion and persistent circulation in Tanzania of a highly virulent p72 genotype II ASFV that is identical to the Georgia 2007/1 isolate in the 3′-end of the B646L gene. An outbreak of ASF in domestic pigs occurred in November 2010 in the Kyela District of the Mbeya region in southwestern Tanzania, which coincided with another outbreak in a neighboring district of Karonga in northern Malawi (Figure, panel A). ASF continued to spread from Mbeya and ultimately reached the neighboring region of Iringa (Ludewa District) in February 2011 through feeding of pigs with swill from Mbeya. By March 2011, ASF had spread to Chunya, Ileje, Mbarali, Rungwe, and Tukuyu districts within Mbeya. The disease spread within the region because of the lack of zoosanitary measures and illegal movement of animals despite the quarantine in place. An outbreak on 1 farm in the Temeke District of the Dar es Salaam region in eastern Tanzania occurred in March 2011 after a farmer obtained pig stock from Mbeya. No further spread of the disease in Dar es Salaam was observed after early diagnosis, removal of affected pigs, and zoosanitary measures. Figure A) Locations in which African swine fever outbreaks occurred during 2010–2012. B–G) Postmortem lesions observed in slaughtered pigs at the Ifakara slaughterhouse of Kilombero District. Postmortem lesions include cutaneous hemorrhage on ... In October 2011, the disease spread to the Sumbawanga District of the Rukwa region through feeding of swill and illegal movement of animals. ASF was reported in February 2012 in Ifakara in the Kilombero District in the Morogoro region, and in July 2012 in the Kilosa District within this region. The disease spread into Kilombero District after 1 farmer purchased pigs for stock from the Iringa region. As of July 2012, ASF was reported again in the Mbeya and Iringa regions, from which it had been eliminated. This unique ASF outbreak in Tanzania persistently circulated for more than a year; previous outbreaks have been sporadic and resolved after shorter durations (8–10). Mortality rates of 100% caused by ASF were recorded in domestic pigs of all ages in all outbreak areas. Affected pigs showed pyrexia and anorexia, dragged their hind legs, and then showed recumbence. In addition, affected animals had severe cutaneous hemorrhages, especially on medial and lateral sides of the pinna, forelimbs above the carpal joint, facial region, scrotum, and mammary glands (Figure, panels B and C). Postmortem lesions included darkening and enlargement of the spleen, severe hemorrhages of mesenteric and gastrohepatic lymph nodes, and hemorrhagic enteritis (Figure, panels D–G). DNA was extracted from spleens of animals that either died of the disease or were killed at slaughterhouses during 2010–2012. The variable 3′-end of the B646L (p72) gene was amplified by using p72U/p72D primers (2) and subjected to automated dideoxynucleotide cycle sequencing by using Big Dye Terminator Cycle Sequencing Kit Version 3.1 (Applied Biosystems, Foster City, CA, USA) in a 24-capillary DNA sequencer (Genetic Analyzer 3500 xL; Applied Biosystems). All obtained ASFV p72 nucleotide sequences (GenBank accession nos. {"type":"entrez-nucleotide","attrs":{"text":"JX391987","term_id":"402759910","term_text":"JX391987"}}JX391987 [TAN/10/Kyela], {"type":"entrez-nucleotide","attrs":{"text":"JX391988","term_id":"402759912","term_text":"JX391988"}}JX391988 [TAN/10/Tukuyu], {"type":"entrez-nucleotide","attrs":{"text":"JX391989","term_id":"402759914","term_text":"JX391989"}}JX391989 [TAN/11/Chunya], {"type":"entrez-nucleotide","attrs":{"text":"JX391990","term_id":"402759916","term_text":"JX391990"}}JX391990 [TAN/11/Ludewa], {"type":"entrez-nucleotide","attrs":{"text":"JX391991","term_id":"402759918","term_text":"JX391991"}}JX391991 [TAN/11/Temeke], {"type":"entrez-nucleotide","attrs":{"text":"JX391992","term_id":"402759920","term_text":"JX391992"}}JX391992 [TAN/12/Ifakara]) were 100% identical. BLAST (http://blast.ncbi.nlm.nih.gov/Blast.cgi) analysis of 2010–2012 ASFV p72 nucleotide sequences from Tanzania in GenBank showed 100% nucleotide identity with the Georgia 2007/1 ASFV isolate (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"FR682468","term_id":"303398661","term_text":"FR682468"}}FR682468) at nt positions 103,594–104,070. The Georgia 2007/1 ASFV isolate was detected in Georgia in 2007 and has caused ASF outbreaks in Armenia, Azerbaijan, and Russia (5). This ASFV isolate belongs to p72 genotype II (5) and clusters with ASFV isolates from Mozambique, Zambia, Madagascar, Mauritius, and Georgia (2,3,7). Although the 2010–2012 outbreak in Tanzania coincided with the outbreak in Malawi in 2010, no ASFV belonging to p72 genotype II has been described in Malawi. ASFV isolates from the 2010 outbreak in Malawi should be sequenced to establish their relatedness to ASFV isolates from the 2010–2012 outbreak in Tanzania and determine an epidemiologic link between these outbreaks. Incursion and persistent circulation of a highly virulent p72 genotype II ASFV identical to the Georgia 2007/1 isolate has implications for transboundary spread of ASF.

Background The neglected tropical diseases, echinococcosis, schistosomiasis and toxoplasmosis are all globally widespread zoonotic diseases with potentially harmful consequences. There is very limited data available on the prevalence of these infections, except for schistosmiasis, in underdeveloped countries. This study aimed to determine the seroprevalence of Echinococcus multilocularis, Schistosoma mansoni, and Toxoplasma gondii antibodies in populations from the Monduli and Babati districts in Tanzania. Methods A total of 345 blood samples were collected from 160 and 185 randomly selected households from Babati and Monduli districts, Tanzania between February and May of 2012 and analyzed them using the enzyme linked immunosorbent assay. The antibodies were determined using the NovaLisa® Toxoplasma gondii IgG, NovaLisa® Schistosoma Mansoni IgG, NovaLisa® Echinococcus IgG and NovaLisa® Toxoplasma gondii IgM kits (Novatec, Germany). Results The seropositivity estimated for E. multilocularis, S. mansoni, and T. gondii IgG was 11.3% (95% confidence interval (CI): 7.96 - 14.6), 51.3% (95% CI: 46.0 - 56.5), and 57.68% (95% CI: 52.5 - 62.9), respectively. The seropositivity for T. gondii IgM was 11.3% (95% CI: 7.96 - 14.6). Living in the Monduli district was found to be the main risk factor for IgG seropositivity for both schistosomiasis (OR =1.94; 95% CI: 1.23 - 3.08; p =0.005) and toxoplasmosis (OR =2.09; 95% CI: 1.31-3.33; p =0.002). Conclusions These results suggest that restricting disease transmission, implementing control measures, and introducing training projects to increase public awareness are imperative, particularly for the Monduli district.

Speed is paramount in the diagnosis of highly infectious diseases, such as foot-and-mouth disease (FMD), as well as for emerging diseases; however, simplicity is required if a test is to be deployed in the field. Recent developments in molecular biology have enabled the specific detection of FMD virus (FMDV) by reverse-transcription loop-mediated isothermal amplification (RT-LAMP), real-time reverse-transcription polymerase chain reaction (RT-qPCR) and sequencing. RT-LAMP enables amplification of the FMDV RNA-dependent RNA polymerase 3D(pol) gene at 63 °C (in the presence of a primer mixture and both reverse transcriptase and Bst DNA polymerase) for 1 h, whilst RT-qPCR amplifies the same gene in approximately 2 h 30 min. In this study, we compared the sensitivity and effectiveness of RT-LAMP against RT-qPCR for the detection of the FMDV 3D(pol) gene in 179 oesophageal-pharyngeal scraping samples (collected by probang) obtained from clinically healthy cattle and buffalo in Malawi, Mozambique and Tanzania in 2010. The FMDV detection rate was higher with RT-LAMP (30.2%; n = 54) than with RT-qPCR (17.3%; n = 31). All samples positive by RT-qPCR (Cq ≤ 32.0) were also positive for the RT-LAMP assay; and both assays proved to be highly specific for the FMDV target sequence. In addition, the VP1 sequences of 10 viruses isolated from positive samples corresponded to the respective FMDV serotypes and genotypes. Our findings indicate that the performance of RT-LAMP is superior to RT-qPCR. Accordingly, we consider this test to have great potential with regard to the specific detection and surveillance of infectious diseases of humans and animals in resource-compromised developing countries.

Peste des petits ruminants (PPR) is an acute viral disease of small ruminants characterised by the sudden onset of depression, fever, oculonasal discharges, sores in the mouth, foul-smelling diarrhoea and death. For many years, in Africa, the disease was mainly confined to West and Central Africa but it has now spread southwards to previously PPR-free countries including Tanzania, Democratic Republic of Congo and Angola. The disease was first reported in Tanzania in 2008 when it was confined to the Northern Zone districts bordering Kenya. Presence of the disease has also been confirmed in southern Tanzania especially Mtwara region. Recently, a suspected outbreak of PPR in Dakawa area, Mvomero district, Morogoro region was reported. Clinical samples (lungs, intestines, lymph nodes, whole blood and sera) from suspected goats (n = 8) and sheep (n = 1) were submitted to Sokoine University of Agriculture for analysis. Molecular diagnosis by amplification of the nucleoprotein gene and the fusion gene of PPR virus (PPRV) using PPRV specific primers was done. Five goats and the sheep were positive for PPRV after performing RT-PCR. To our knowledge, this is the first report confirming the presence of PPR in the Mvomero district of the Morogoro region, Tanzania. Hence, more efforts should be put in place to prevent the spread of PPR in Tanzania.

Foot-and-mouth disease (FMD) is an acute, highly contagious viral infection of domestic and wild cloven-hoofed animals. It is known to be endemic in Zambia, with periodic outbreaks occurring in different geographical areas of the country. This study was conducted to investigate the presence of FMD virus (FMDV) in reported FMD-suspected cases in cattle from the Kazungula and Mbala districts of Zambia. Sixty epithelial tissues or oesophageal-pharyngeal (OP) scrapings (probang samples) were collected from Mbala (n = 51) and Kazungula (n = 9) and examined for FMDV. The FMDV viral RNA and serotypes were examined by realtime reverse transcription polymerase chain reaction (qRT-PCR) and antigen Enzyme- linked immunosorbent assay (ELISA), respectively. Twenty-two samples (36.7%) were positive for the FMDV genome by qRT-PCR with Cycle threshold (Ct) values ranging from 13 to 31. The FMDV-positive samples from epithelial tissues showed relatively higher Ct values compared to those obtained from OP scrapings, irrespective of geographical location. Forty percent (40%; n = 4) of epithelial tissues from Mbala were serotyped into SAT 2 serotype by antigen ELISA. Kazungula samples were serotyped into SAT 1. These findings indicated that Mbala and Kazungula districts had FMD outbreaks in 2012 that were ascribed to at least FMDV serotype SAT 2 and SAT 1 field strains. Furthermore, regular interaction between buffalos from the Mosi-o Tunya Park and domestic animals from surrounding areas could contribute to the occurrence of regular FMD outbreaks in Kazungula, whilst the uncontrolled animal movements across borders between Mbala and Nsumbawanga could be responsible for disease outbreaks in Mbala. In-depth molecular biological studies, including sequencing and phylogeny of the viruses, should be conducted to elucidate the complex epidemiology of FMD in Zambia, thereby providing valuable information needed for the rational control strategy of FMD in Zambia and neighbouring countries.

This study was conducted to determine the spatiotemporal distribution of foot-and-mouth disease (FMD) virus (FMDV) serotypes and evaluate the awareness of livestock keepers about FMD in Tanzania. An observational prospective study involving serological analysis, FMDV antigen detection and questionnaire survey was carried out in the lake zone of Tanzania. Seroprevalence of antibodies to the nonstructural protein 3ABC of FMDV and serotype-specific antigen detection were investigated by using SVANOVIR® FMDV 3ABC-Ab ELISA and indirect-sandwich ELISA (sELISA), respectively, whilst a structured questionnaire was used to evaluate the awareness of livestock keepers about FMD. During the period of 2010–2011, both serum and tissue (foot-and-mouth epithelia) samples were collected from cattle suspected of FMD in 13 districts of the four regions of the lake zone. A total of 107 (80.5%) out of 133 tested serum samples were seropositive to nonstructural protein 3ABC, with at least one sample being positive from all 10 districts screened. Fifteen (53.6%) out of 28 tissue epithelial samples collected from FMD cases in eight districts during the course of this study were positive to serotype O FMDV antigen. Of these eight districts, serotype O FMDV antigens were detected from seven districts and no other serotypes were recovered from animal samples screened. Questionnaire surveys in six districts indicated that livestock keepers in the lake zone were aware of the clinical manifestations (26/29 = 90.0%) and economic impact (23/29 = 79.0%) of FMD in the region. The questionnaire data showed that FMD outbreaks often occurred after rainy seasons (22/29 = 75.9%), with the highest peaks predominantly occurring just after the long rains in May and June, and at the end of the short rains in November and December of each year. The spatial distribution of the FMD cases suggested that serotype O virus exposure was the only widespread cause of the 2010–2011 outbreaks in the lake zone.