Browsing by Author "Misinzo, G."
Now showing 1 - 20 of 20
- Results Per Page
- Sort Options
Item Antigenic differences among porcine circovirus type 2 strains, as demonstrated by the use of monoclonal antibodies(Journal of General Virology, 2008) Lefebvre, D. J.; Costers, S.; Doorsselaere, J. V.; Misinzo, G.; Delputte, P. L.; Nauwynck, H. J.This study examined whether antigenic differences among porcine circovirus type 2 (PCV-2) strains could be detected using monoclonal antibodies (mAbs). A subtractive immunization protocol was used for the genotype 2 post-weaning multisystemic wasting syndrome (PMWS)-associated PCV-2 strain Stoon-1010. Sixteen stable hybridomas that produced mAbs with an immunoperoxidase monolayer assay (IPMA) titre of 1000 or more to Stoon-1010 were obtained. Staining of recombinant PCV-2 virus-like particles demonstrated that all mAbs were directed against the PCV-2 capsid protein. Cross-reactivity of mAbs was tested by IPMA and neutralization assay for genotype 1 strains 48285, 1206, VC2002 and 1147, and genotype 2 strains 1121 and 1103. Eleven mAbs (9C3, 16G12, 21C12, 38C1, 43E10, 55B1, 63H3, 70A7, 94H8, 103H7 and 114C8) recognized all strains in the IPMA and demonstrated neutralization of Stoon-1010, 48285, 1206 and 1103, but not VC2002, 1147 and 1121. mAbs 31D5, 48B5, 59C6 and 108E8 did not react with genotype 1 strains or had a reduced affinity compared with genotype 2 strains in the IPMA and neutralization assay. mAb 13H4 reacted in the IPMA with PMWS-associated strains Stoon-1010, 48285, 1206 and VC2002, and the porcine dermatitis and nephropathy syndrome-associated strain 1147, but not with reproductive failure-associated strains 1121 and 1103. mAb 13H4 did not neutralize any of the tested strains. It was concluded that, despite the high amino acid identity of the capsid protein (¢91 %), antigenic differences at the capsid protein level are present among PCV-2 strains with a different genetic and clinical background.Item Binding and entry characteristics of porcine circovirus 2 in cells of the porcine monocytic line 3D4/31(2005) Misinzo, G.; Meerts, P.; Bublot, M.; Mast, J.; Weingartl, H. M.; Nauwynck, H. J.Porcine circovirus 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome and reproductive problems in pigs. Cells of the monocyte/macrophage lineage are important target cells in PCV2-infected pigs, but the method of binding and entry of PCV2 into these cells is unknown. Therefore, binding and entry of PCV2 to the porcine monocytic cell line 3D4/31 were studied by visualization of binding and internalization of PCV2 virus-like particles (VLPs) by confocal microscopy and chemical inhibition of endocytic pathways (clathrin- and caveolae-mediated endocytosis and macropinocytosis), followed by evaluation of the level of PCV2 infection. It was shown that PCV2 VLPs bound to all cells, with maximal binding starting from 30 min post-incubation. Bound PCV2 VLPs were internalized in 47±5?0% of cells. Internalization was continuous, with 70?5±9?7% of bound PCV2 VLPs internalized at 360 min post-incubation. Internalizing PCV2 VLPs co-localized with clathrin. PCV2 infection was decreased significantly by chemical inhibitors that specifically blocked (i) actin-dependent processes, including cytochalasin D (75?5±7?0% reduction) and latrunculin B (71?0±3?0% reduction), and (ii) clathrin-mediated endocytosis, including potassium depletion combined with hypotonic shock (50?2±6?3% reduction), hypertonic medium (56?4±5?7% reduction), cytosol acidification (59?1±7?1% reduction) and amantadine (52?6±6?7% reduction). Inhibiting macropinocytosis with amiloride and caveolae-dependent endocytosis with nystatin did not decrease PCV2 infection significantly. PCV2 infection was reduced by the lysosomotropic weak bases ammonium chloride (47?0±7?9% reduction) and chloroquine diphosphate (49?0±5?6% reduction). Together, these data demonstrate that PCV2 enters 3D4/31 cells predominantly via clathrin-mediated endocytosis and requires an acidic environment for infection.Item Cell tropism and entry of porcine circovirus 2(Elsevier, 2011-11-11) Nauwynck, H.J.; Sanchez, R.; Meerts, P.; Lefebvre, D.J.; Saha, D.; Huang, L.; Misinzo, G.Porcine circovirus 2 (PCV2) may induce reproductive failure (return to oestrus, embryonic death, mum- mification, weak- and stillborn piglets) and postweaning multisystemic wasting syndrome (PMWS). Furthermore, it may modulate the immunity in such a way that it aggravates the outcome of many bacterial and viral infections. In the present paper, the cellular tropism and entry of PCV2 are described and linked with the pathological and clinical consequences.Item Clinico-pathological findings of the 2011 outbreak of peste des petits ruminants (PPR) in Tandahimba district, southern Tanzania(Research Opinions in Animal and Veterinary Sciences, 2012) Matondo, R. B.; Muse, E. A.; Karimuribo, E. D.; Misinzo, G.; Albano, M. O.; Gitao, G. C.Although PPR outbreaks were reported in Northern Tanzania since 2008, there has been no description of the clinical or pathological manifestation of the disease, an important criterion in guiding veterinarians and farmers on proper recognition and diagnosis of the disease. A study was therefore conducted to investigate and describe clinical signs and pathological lesions associated with 2011 Peste des petits ruminants (PPR) outbreak in goats and sheep in Tandahimba district located in Southern Tanzania. The investigation involved taking history and conducting clinical examination of PPR suspected cases (25 goats and 3 sheep) in the study district which had neither a history of vaccination against PPR nor previous illness due to PPR. This work was complemented by collection of pathological samples and specimens for laboratory examination. A detailed post-mortem was performed on three sacrificed animals followed by collection of specimens including lungs, liver, spleen and lymph nodes for histopathological examination. Clinical samples from 30 animals which included swabs from ocular, nasal and mouth lesions were also collected for confirmation of PPR through detection of PPR ribonucleic acid using reverse transcription polymerase chain reaction (RT-PCR). Clinical examinations of the cases showed signs suggestive of PPR including severe depression, high fever (41oC), anorexia, muco-pulurent nasal discharge, erosive and necrotic stomatitis, mild diarrhoea and skin nodules. Post mortem examination showed evidence of pneumonia including lung congestion and consolidation, increased thickness of inter-alveolar walls, moderate infiltration of inflammatory cells in bronchiolar subepithelial and perivascular layers. Overall 56.7% of the samples (n=30) tested were positive for PPR by RTPCR. This study has confirmed and described the presence of PPR in southern Tanzania. A more detailed study including other districts is recommended to provide more information regarding the magnitude and factors associated with PPR in Southern Tanzania.Item Correlation between the presence of neutralizing antibodies against porcine circovirus 2 (PCV2) and protection against replication of the virus and development of PCV2-associated disease(2006-01-30) Meerts, P.; Misinzo, G.; Lefebvre, D.; Nielsen, J.; Bøtner, A.; Kristensen, C. S.; Nauwynck, H. J.Background: In a previous study, it was demonstrated that high replication of Porcine circovirus 2 (PCV2) in a gnotobiotic pig was correlated with the absence of PCV2-neutralizing antibodies. The aim of the present study was to investigate if this correlation could also be found in SPF pigs in which PMWS was experimentally reproduced and in naturally PMWS-affected pigs. Results: When looking at the total anti-PCV2 antibody titres, PMWS-affected and healthy animals seroconverted at the same time point, and titres in PMWS-affected animals were only slightly lower compared to those in healthy animals. In healthy animals, the evolution of PCV2-neutralizing antibodies coincided with that of total antibodies. In PMWS-affected animals, neutralizing antibodies could either not be found (sera from field studies) or were detected in low titres between 7 and 14 DPI only (sera from experimentally inoculated SPF pigs). Differences were also found in the evolution of specific antibody isotypes titres against PCV2. In healthy pigs, IgM antibodies persisted until the end of the study, whereas in PMWS-affected pigs they quickly decreased or remained present at low titres. The mean titres of other antibody isotypes (IgG1, IgG2 and IgA), were slightly lower in PMWS-affected pigs compared to their healthy group mates at the end of each study. Conclusion: This study describes important differences in the development of the humoral immune response between pigs that get subclinically infected with PCV2 and pigs that experience a high level of PCV2-replication which in 3 of 4 experiments led to the development of PMWS. These observations may contribute to a better understanding of the pathogenesis of a PCV2- infection.Item Current epidemiological assessment of Bancroftian Filariasis in Tanga region, Northeastern Tanzania(Hindawi Publishing Corporation, 2016) Mshana, H. J.; Baraka, V.; Misinzo, G.; Makunde, W. H.Tanzania started a countrywide lymphatic filariasis elimination programme in 2000 adopting the mass drug administration (MDA) strategy.The drug used for the programme was the combination of ivermectin and albendazole. However, there is limited information on the current epidemiological trend of the infections, where MDA implementation is ongoing. The present study aimed at assessing the current status of Bancroftian filariasis infection rate and morbidity where MDA has been distributed and administered for over eight rounds. Methodology. The study was a cross-sectional descriptive study involving 272 individuals (>18 years) from endemic communities in Tanga region where MDA has been implemented. Clinical, sociodemographic, and circulating filarial antigen (CFA) test was undertaken usingimmune chromatographic card test according to themanufacturer’s instructions. Results. A total of 472 individuals were screened: 307/472 (65.1%) were males while 165/472 (34.9%) were females. The overall prevalence of CFA was 5.51%, that of hydrocoele was 73.2%, and that of lymphoedema was 16.0%. The prevalence of hydrocoele combined with lymphoedema was 5.5%. Conclusion. Our findings demonstrate a considerable reduction in filarial infection. However, there is clear evidence of ongoing transmission despite the 8 rounds of MDA. It is unlikely that the annual MDA would interrupt filarial transmission; therefore, additional strategies are needed to accelerate lymphatic filariasis control and elimination.Item Distribution and diversity of mosquitoes and the role of Aedes in the transmission of arboviruses in selected districts of Tanzania(2017-12-25) Patrick, N. P.; Kinimi, E.; Shayo, M/ J.; Ang eenyi, S. O.; Weyer, P.; van Vuren, P. J.; Paweska, J. T.; Mboera, L. E. G.; Rweyemamu, M. M.; Misinzo, G.; Kasanga, C. J.Arboviruses belong to various families of viruses that are transmitted by arthropods, mainly mosquitoes and often cause diseases in humans. The objective of this study was to determine mosquito diversity and transmission of arboviruses by Aedes in selected ecosystems in Tanzania. Adult mosquitoes were collected from rural and urban settings using carbon dioxide-baited CDC light traps, Biogent sentinel traps, and the Mosquito Magnet traps. Reverse Transcription-Polymerase Chain Reaction assay was performed on pooled adult Aedes mosquitoes to detect the presence of Chikungunya, Dengue, Rift Valley fever (RVF) and Yellow fever (YF) viruses. A total of 42, 77 mosquitoes belonging to five genera (Aedes, Anopheles, Culex, Mansonia and Mimomyia) and 18 species were collected. Culex accounted for the largest (62.7%; n= 2,682) proportion of the mosquitoes while Anopheles for the lowest proportion (5.7%; n=245). Of the total mosquitoes collected, Culex quinquefasciatus accounted for more than a half (53.4%; n=2692), followed by Aedes aegypti 12.1% (n=520). Of the 34 adult Ae. aegypti pools tested, arboviruses were detected in 33(97%) pools. Dengue virus was detected in 47.6% (10/ 21) pools which tested positive for Flaviruses. Chikungunya virus was detected in 30% (3/ 10) pools which were positive for Alphavirus genera. Of 2 pools tested positive for Bunyavirus genus, Rift Valley fever virus was detected in 1 pool (50%). The presence of various mosquito vectors and detection of arboviruses in aedes mosquitoes leave the population of Tanzania at great risk of transmission of different pathogens and highlight a need for vector control measures in the country.Item Epidemiological investigation into the introduction and factors for spread of Peste des Petits Ruminants, southern Tanzania(2012) Muse, E. A.; Karimuribo, E. D.; Gitao, G. C.; Misinzo, G.; Mellau, L. S. B.; Msoffe, P. L. M.; Swai, E. S.; Albano, M. O.A study was carried out to confirm and identify sources and elucidate factors associated with the introduction of Peste des Petits Ruminants (PPR) in southern Tanzania. This study was conducted in Tandahimba and Newala districts of Mtwara region following suspected outbreak of PPR in the area. Qualitative data were collected using semi-structured questionnaires and in-depth interviews of key informants who included goat and sheep owners with suspected cases of PPR and animal health service providers as well as local administrative authority. Additionally, 216 serum samples and 28 swabs were collected for serological and virological laboratory disease confirmation. The results show that PPR was first introduced in Likuna village of Newala district in February 2009 through newly purchased goats from the Pugu livestock market located about 700 km in the outskirts of Dar es Salaam city. Factors which contributed to spread of PPR included communal grazing and the cheap prices of sick animals bought by livestock keepers for slaughtering in other villages. Laboratory findings confirmed presence of PPR in the area by RT-PCR and serological analysis revealed that seroprevalence was 31%. These findings have confirmed, for the first time, introduction of PPR in southern Tanzania. The presence of PPR poses high risk of southward spread of the disease to other southern African countries in the SADC region thus calling for concerted and collaborative efforts in prevention and control of the disease to avoid losses. Further elaborate studies on the spread, prevalence and risk factors associated with the disease should urgently be investigated.Item Evidence of chikungunya virus infection among febrile patients seeking healthcare in selected districts of Tanzania(Taylor & Francis, 2018) Kinimi, E.; Shayo, M. J.; Patrick, B. N.; Angwenyi, S. O.; Kasanga, C. J.; Weyer, J.; Vuren, P. J.; Paweska, J. T.; Mboera, L. E.G.; Misinzo, G.Introduction: Chikungunya virus (CHIKV) infection is an emerging mosquito-borne disease that has been associated with frequent epidemics in the world. However, there is a dearth of information on its magnitude and associated risk factors in Tanzania. Objective: A study was conducted to determine seroprevalence of CHIKV among febrile patients seeking medical care at health facilities in Karagwe, Sengerema, Kilombero and Kyela districts. Methods: Structured questionnaires were administered and 728 serum samples were col- lected between May and June, 2015 and tested for the presence of CHIKV-IgM and IgG- specific antibodies using Enzyme-linked immunosorbent assay. Results and discussion: The common clinical characteristics exhibited by outpatients were fever, headache and joint pains (100%, 70%, and 68.3% respectively). Out of 728 outpatients screened for CHIKV, 105 (14%) tested CHIKV IgG positive whilst 11 (1.5%) tested CHIKV IgM positive. Chikungunya seropositivity was significantly higher than previously reported in Tanzania. The most affected age group was 20–29 years. Our results indicate that CHIKV infection is prevalent and contributes to the burden of febrile illnesses in Tanzania. The seroprevalence varies between districts, reflecting variation in mosquito vector transmission dynamics in different parts of the country. Received 23 September 2018 Accepted 19 November 2018 KEYWORDS Chikungunya; seroprevalence; febrile illness; mosquito-borne; Tanzania Abbreviations: CHIKV: Chikungunya virus; EDTA: Ethylenediaminetetraacetic acid; ELISA: Enzyme-linked immunosorbent assay; IgG: Immunoglobulin G; IgM: Immunoglobulin M; NIMR: National Institute for Medical Research; RU: Relative Units; SACIDS: Southern African Centre for Infectious Disease Surveillance; USA: United States of AmericaItem Genetic characterization of African swine fever viruses from a 2008 outbreak in Tanzania(2011-02) Misinzo, G.; Magambo, J.; Masambu, J.; Yongolo, M. G.; Doorsselaere, J. V.; Nauwynck, H. J.Outbreaks of African swine fever (ASF) have been reported in the past from several countries in sub-Saharan Africa. The aim of this study was to genotype ASF viruses (ASFVs) from the 2008 outbreak in Morogoro and Dar es Salaam regions of Tanzania. Tissue samples from domestic pigs that died as a result of severe haemorrhagic disease were collected and analysed with PCR and genome sequencing methods using ASFV-specific primer sets. Nucleotide sequence data were obtained for the B646L (p72), E183L (p54) and the variable region of the B602L gene sequences. Phylogenetic analyses based on DNA sequences showed that the 2008 Tanzanian isolates belonged to p72 genotype XV and clustered together with those derived from the 2001 outbreak in Tanzania. Analysis of the tetrameric amino acid repeat regions within the variable region of the B602L gene showed that the repeat signature of the 2008 Tanzanian ASFV was unique and contained three novel tetramers (U = NIDT/NTDT and X = NTDI). Epidemiological investigation suggested that transportation of live pigs continues to play an active role in the epidemiology of ASF in Tanzania. It is recommended that future control of ASF spread in Tanzania should focus on the early detection and confirmation of the disease, prompt institution of quarantine measures, culling and proper disposal of infected and in-contact animals and decontamination of affected premises.Item Increased porcine circovirus type 2 replication in porcine leukocytes in vitro and in vivo by concanavalin A stimulation(Veterinary Microbiolog, 2008-05-05) Lefebvre, D. J.; Meerts, P.; Costers, S.; Misinzo, G.; Barbe, F.; Reeth, K. V.; Nauwynck, H. J.Previously, it was shown that modulation of the immune system enhances porcine circovirus type 2 (PCV2) replication in pigs. In the present study, the effect of the mitogen concanavalin A (ConA) on PCV2 replication was investigated. Since ConA induces T-lymphocyte activation and initiates the production of interferon-gamma (IFN-g), a cytokine that enhances PCV2 replication in porcine epithelial and monocytic cell lines in vitro, it was examined if the effects observed with ConA were mediated by IFN-g. In an in vitro study, ConA but not IFN-g enhanced PCV2 replication in peripheral blood mononuclear cells (PBMC). Up to 2.08% and 0.96% of PBMC were antigen positive for PCV2 strains 1121 and Stoon-1010, respectively, and a low virus production was observed. PCV2-infected PBMC were identified as CD4+ (40%), CD8+ (54%) and IgM+ (11%). In a subsequent in vivo study, caesarean-derived colostrum-deprived piglets were injected with ConA or IFN-g 12 h before inoculation and every 3 days for 9 days after inoculation with strain 1121. PCV2 was isolated from inguinal lymph node biopsies from 10 days post-inoculation (dpi) in ConA-treated pigs and from 15 dpi in non-treated and IFN-g-treated pigs. ConA increased PCV2 replication levels, but disease was not observed. Half of the ConA-treated and IFN-g-treated pigs showed a delayed humoral immune response, but this delay did not result in increased PCV2 replication in these pigs. These experiments demonstrated that ConA enhances PCV2 replication in PBMC in vitro and in lymphoid tissues in vivo. # 2008 Elsevier B.V. All rights reserved.Item Increased yield of porcine circovirus-2 by a combined treatment of PK-15 cells with interferon-gamma and inhibitors of endosomallysosomal system acidification(2007-12-17) Misinzo, G.; Delputte, P. L.; Lefebvre, D. J.; Nauwynck, H. J.Treatment of porcine kidney (PK-15) cells with either interferon-gamma (IFN-g) or endosomallysosomal system acidification inhibitors increases replication of porcine circovirus type 2 (PCV2). In the present study, the effect of a combination of these treatments on the number of infected cells and virus yield was tested. The number of PCV2 (Stoon-1010)-infected PK-15 cells increased in cells treated with ammonium chloride (445 39% increase), IFN-g (446 8%), ammonium chlorideþ IFN-g (1721 283%), chloroquine diphosphate (1037 121%), chloroquine diphosphateþIFN-g (2199 255%), monensin (950 178%) and monensinþIFN-g (1948 60%). Combined IFNg and endosomal-lysosomal system acidification inhibitors increased PCV2 yield by up to 50 times compared to untreated PK-15. This augmented virus replication in PK-15 cells may be helpful in the production of PCV2 vaccines.Item Partial genetic characterization of peste des petits ruminants virus from goats in northern and eastern Tanzania(Transboundary and Emerging Diseases, 2014) Kgotlele, T.; Macha, E. S.; Kasanga, C. J.; Kusiluka, L. J. M.; Karimuribo, E. D.; Doorsselaere, J. V.; Wensman, J. J.; Munir, M.; Misinzo, G.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.Item Peste des Petits Ruminants (PPR) outbreak in southern, Tanzania(RUFORUM, 2012) Muse, E. A.; Matondo, R. B.; Karimuribo, E. D.; Misinzo, G.; Mellau, L. S. B.; Msoffe, P. L. M.; Albano, M. O.; Gitao, G. C.Peste des petits ruminants (PPR) was first confirmed in Tanzania in 2008, however description of clinical or pathological signs was not carried out although this is important to assist quick identification and reporting of PPR cases by both livestock keepers and field-based animal health workers. A study was therefore conducted to investigate and describe clinical signs and pathological lesions associated with suspected PPR cases in southern Tanzania. It involved history taking and clinical examination of suspected cases of 25 goats and 3 sheep. Post- mortem examination of some cases was performed followed by collection of specimens for histopathological examination. Swabs were also collected for confirmation of PPR by detecting ribonucleic acid using reverse transcription polymerase chain reaction (RT-PCR). Serum samples were analysed using competitive enzyme linked immunosorbent assay (cELISA). Severe depression, high fever, anorexia, muco-pulurent nasal discharge, erosive and necrotic stomatitis, mild diarrhoea and skin nodules were major signs suggestive of PPR. Post mortem examination showed evidence of pneumonia including lung congestion and consolidation. RT-PCR confirmed presence of the PPR virus in samples and serum antibodies showed seroprevalence of 31%.Item Porcine reproductive and respiratory syndrome virus entry into the porcine macrophage(Journal of General Virology, 2010) Breedam, W. V.; Delputte, P. L.; Van Gorp, H.; Misinzo, G.; Vanderheijden, N.; Duan, X.; Nauwynck, H. J.Porcine reproductive and respiratory syndrome virus (PRRSV) emerged in the late 1980s and rapidly became one of the most significant viral pathogens in the swine industry. In vivo, the virus shows a very narrow cell tropism and targets specific subsets of porcine macrophages. The entry of PRRSV into its host cell is the first crucial step in infection and has been the focus of many fundamental studies. This review provides a comprehensive overview of the current knowledge on PRRSV entry into the porcine macrophage, covering virus binding, internalization and genome release, and integrates these findings into a general model of the entry process.Item Preliminary investigation on presence of peste des petits ruminants in Dakawa, Mvomero district, Morogoro region, Tanzania(2014) Kgotlele, T.; Kasanga, C. J.; Kusiluka, L. J. M.; Misinzo, G.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, foulsmelling 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.Item Replication kinetics of different porcine circovirus 2 strains in PK-15 cells, fetal cardiomyocytes and macrophages(2005) Meerts, P.; Misinzo, G.; McNeilly, F.; Nauwynck, H. J.In this in vitro study, the replication kinetics of porcine circovirus type 2 (PCV2) in porcine alveolar macrophages (PAM) and fetal cardiomyocytes (FCM), two target cells in vivo, was compared with that in PK-15 cells. Cultures were inoculated with either the postweaning multisystemic wasting syndrome (PMWS)-associated strain Stoon-1010 or the abortion-associated strain 1121. Viral proteins were visualized and virus production was determined. In PK-15 cells, the capsid protein was expressed between 6 and 12 hours post inoculation (hpi), it relocated to the nucleus between 12 and 24 hpi. At that time, Rep protein was also detected in the nucleus. This sequence of events also occurred in FCM and PAM but nuclear localized antigens appeared later (48 hpi) and in a lower percentage of cells. In PAM, clear differences in susceptibility were seen between pigs. In PAM from two out of five tested pigs, nuclear localized antigens were not detected, whereas in PAM from three other pigs they were seen in up to 20% of the antigen-positive cells. Virus production was observed in PK-15 but not in PAM or FCM cultures. In a second study, the replication kinetics of seven different PCV2 strains were compared in PK- 15 cells. It was shown that the two abortion-associated strains had a different replication kinetics in comparison with PMWS or porcine dermatitis and nephropathy syndrome associated strains. With the abortion-associated strains, a higher number of infected cells was observed at 24 hpi and the percentage of infected cells with nuclear localised antigens was lower compared to that of other strains.Item Serological evidence of chikungunya and malaria co-infection among febrile patients seeking health care in Karagwe district, Tanzania(2018-10) Kinimi, E.; Patric, P. N.; Misinzo, G.Background: Chikungunya is an emerging mosquito-borne viral illness of major public health concern and is becoming a common infection in many geographical areas of Tanzania. This study was carried out to determine the incidence of malaria and chikungunya infections among febrile patients seeking medical care in Karagwe district, Tanzania. Methods: Febrile patients were enrolled into the study at Nyakahanga district designated hospital and Kayanga heath centre in May and June 2015. Questionnaires were administered to collect clinical and socio-demographic characteristics of patients. All participants were tested for malaria using malarial rapid diagnostic test and those tested positive by mRDT were confirmed by microscopy. Both outpatients tested malaria positive and negative were further screened for immunoglobulin M (IgM) and G (IgG) antibodies for chikungunya using enzyme-linked immunosorbent assay. Results: A total of 400 febrile patients were enrolled in the study. Out of 400 febrile outpatients tested for malaria, 116 (28.75%) tested positive with mRDT. Microscopy confirmed presence malaria parasites in 112 (96.55%) of the malaria RDT-positive. The overall seroprevalence of chikungunya infection was 24.25% (97/400). Out of those chikungunya seropositive subjects, 89 (91.75%) had no malaria. Co-infection rate of chikungunya and malaria was found to be 7.14% (8/112). Conclusions: Our findings confirmed the existence of chikungunya and malaria co-infection among febrile patients seeking health care in Karagwe district. Chikungunya should be considered in the differential diagnosis of malaria for appropriate case management and in order to monitor the public health burden and to inform possible preventative and control measures.Item Seroprevalence of leptospira infection from agro pastoralist communities in Katavi ecosystem, Tanzania(2014-04-14) Muller, S. K.; Asenga, L. F.; Matemba, L.; Misinzo, G.; Kazwala, R.Background: Leptospirosis is a neglected zoonotic disease of worldwide public health importance which affects both humans, domestic and wildlife. Our previous study in Katavi ecosystem showed that prevalence of leptospirosis in livestock was 28%. This predisposes the agro-pastoralist communities at high risk of the diseases. Microscopic agglutination test (MAT) is the gold standard technique for diagnosis of Leptospirosis. This cross sectional study intended to provide serological data for the circulating Leptospira species in Western part of Tanzania. Methods & Materials: 265 blood samples from healthy partici- pants living in Katavi ecosystem were collected in plain vacutainer tubes, centrifuged for sera collection and stored in liquid nitro- gen. Urine samples were also collected and cultured in Fletcher Leptospira media for isolation of live organism. To be certain of other causes of febrile illness in the region; Screening tests for malaria and brucellosis (mRDT and Rose Bengal) were done respec- tively. All samples were processed at Mpanda District Hospital and transported to Sokoine University of Agriculture (SUA) for further analysis. (MAT) was done using six known Leptospira interrogans serovars: Pomona, Icterohaemorrhagiae, Ballum, Tarassovi, Grippotyphosa and Hardjo. Starting with Serovar Grippothy- posa. Resulting agglutination titers were read using dark field microscopy. Results: Confirmed leptospirosis was outlined as a ≥ 4-fold increase in microscopic agglutination test (MAT) titer. Out of 265 participants, 3.8% were exposed to Leptospira Serovar Gryppoty- phosa, 5.6% (15) had significant positive titer for Leptospira Serovar Gryppotyphosa. Apart from Leptospirosis; 13.8% of participants were malaria positive and 1.4% were brucellosis positive. Among those negative for malaria and brucellosis; 13 (5.7%) had high pos- itive titer for Leptospirosis. 2 participants were co- infected with malaria and Leptospirosis. This is just preliminary results, results of other serovars will be completed in December 2013 Conclusion: This study detected the circulating Leptospira Serovars in agro-pastoralist communities living in Katavi ecosys- tem. Serovar Gryppotyphosa is among the circulating Leptospira serovars in Katavi region. This information is significant for better understanding of epidemiology of Leptospirosis in Katavi Region. Molecular techniques like PCR, whole order sequencing ought to be thought of in future studies.Item Studies of brucellosis in lactating cows in Babati district, Tanzania(Tanzania Veterinary Association, 2017-12-07) Kayombo, G.; Makingi, G.; Nonga, H. E.; Misinzo, G.; Kazwala, R. R.The present cross-sectional study was carried out to determine prevalence and risk factors for transmission of brucellosis in lactating cows in Babati district. Rose Bengal plate test (RBPT), buffered acidified plate test (BAPA), competitive enzyme-linked immunosorbent assay (c-ELISA) and polymerase chain reaction (PCR) tests were used in this study to determine the presence of antibodies against Brucella and Brucella genome. Milk and blood samples from 192 randomly selected lactating cows were collected. Furthermore, questionnaires were administered to 66 milk producers to determine the risk factors for the transmission of brucellosis in between animal populations. The RBPT and BAPA results showed 4.7% (nine cows) and 5.2% (10 cows) seroprevalence, respectively. When RBPT and BAPA positive samples were tested using c-ELISA for serologic confirmation, only eight cows (4.2%) turned out to be positive. The milk samples from eight cows that were positive for Brucella antibodies using c-ELISA were tested for the presence of Brucella DNA using PCR. Three out of the eight milk samples were positive for Brucella abortus indicating shedding of Brucella in milk. Analysis of risk factors for transmission of brucellosis by Fisher‘s exact test or Chi-square showed that livestock mixing with different herds (P=0.0097, OR=11.3333), farming system of cattle (P=0.0400, OR=3.9474), breed of cattle (P=0.0284, OR=1.9860), herd size of cattle (P=0.0030, OR=1.9537) and movement of animals through selling and purchasing (P=0.0500, OR=5.0588) were statistically associated with Brucella positivity. This study provides evidence of brucellosis in lactating cows of Babati district and shedding of Brucella in milk. Institution of appropriate control measures including public health education, surveillance of animals accompanied with removal of positive cases according to laws and immunisations of cattle are highly recommended.