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Item Taenia saginata cysticercosis in cattle in Tanzania(Sokoine University of Agriculture, 1993) Maeda Godfrey ElikaliaA review on the epidemiology and diagnosis of human taeniasis and bovine cysticercosis with special reference to East Africa is given. All humans are susceptible to the adult tapeworm, T. saginata. Cattle, is the predominant intermediate host, young animals being more susceptible than older ones. Susceptibility of a few wild ruminants makes them potential reservoirs of infection. Characteristics of T. saginata, including a long life span, a high reproductive potential, spontaneous shedding of proglottids from human carriers, occult nature of the cysticerci in cattle, etc, contribute to maintenance of infections. Dispersion and survival of T. saginata eggs are determined by sanitation, personal hygiene, coprophagous animals and weather conditions. Prevalence rates of cysticercosis in cattle reported from various countries depend on local meat hygiene legislations, efficiency of meat inspection, and record keeping in the abattoirs. In this thesis, factors promoting the spread of T. saginata infections in Tanzania were investigated, by interview information from 105 cattle owners, belonging to Masai, Arusha, Iraqw, Gogo and Rangi ethnic groups, from 52 villages in Tanzania. Taeniasis and cysticercosis were reported as common problems among members of the ethnic groups interviewed. Lack of awareness of the source of human tapeworms, scarcity of medicines against taeniasis, habits of consuming raw beef, consumption of uninspected meat, defaecation in the bush, were some of the outstanding factors elucidated. The details of this investigation are discussed and it is concluded that some of the observations need confirmation through further studies in the villages of the ethnic groups concerned. Prevalence rates of taeniasis recorded at Mbulu Hospital were 10% in 1990 and 21% in 1991. In a separate investigation, cysticercosis was detected in 52 (10.5%) out of 496 cattle X xi slaughtered in municipal abattoirs of Arusha, Dodoma, Iringa, Morogoro and Mpwapwa districts in Tanzania during the period between June 1991 and November 1991. The prevalence rates in the individual abattoirs were 16.7% (Arusha), 8% (Dodoma), 9.6% (Iringa), 6.5% (Morogoro) and 7.6% (Mpwapwa). The overall annual prevalence rates of cysticercosis in the Dodoma and Iringa regions were lower than the rates officially recorded in these abattoirs. The results are discussed in relation to sources of infection, endemicity of cysticercosis, efficiency of meat inspection, and records keeping in the abattoirs in the country. In a detailed examination, anatomical distribution of cysticerci of T. saginara (864) was analyzed in zebu cattle at Morogoro abattoir in Tanzania. The cysticerci were found located in the examined tissues preferentially in the following order: heart, M. triceps brachii, tongue, M. psoas and masseter muscles. The liver had relatively high numbers of cysts and is thus considered an important predilection site. Examination of the predilection sites for detecting carcasses positively infected with cysticerci of T. saginara revealed the following efficiency: 80% (heart), 80% (M. triceps brachii), 60% (masseter muscles), 60% (tongue) and 53% (liver). However examination of the heart, M. triceps brachii and the liver together detected all infected carcasses. Most (60%) of the infected carcasses had viable cysticerci. In conclusion, T. saginara taeniasis and cysticercosis are major, but underestimated problems in Tanzania. The diagnostic methods of cysticercosis in cattle and taeniasis in humans all have inherent limitations. But nevertheless, their sensitivity may be significantly be improved and the usual control may be more efficient. The various chapters of the thesis include discussions on ways to improve public control measures and educate consumers and herdsmen.Item Urinary tract infection: The role of canine transmissible venereal tumour(Wiley Online Library, 1996) Batamuzi, E. K.; Kristensen, F.The role of transmissible ‘venereal tumours in the pathogenesis of urinary tract infection in dogs was investigated in 86 dogs. Fifty-five had transmissible venereal tumours, and the remaining 31 animals were used as controls. A thorough clinical examination of the external genitalia was carried out in each case. In the dogs with transmissible venereal tumours, the sites of attachment were recorded. Urine samples were taken by cystocentesis and the external genitalia swabbed; the samples were cultured for bacteria using standard methods. Tumours were found on the prepuce and other parts of the penis in male dogs; in bitches they were found in the vagina, vestibule or the vestibulovaginal junction. Dogs with transmissible venereal tumours were found to be at a high risk of having bacteriuria (odds ratio [OR] = 7.04). Obliteration of the urethral orifice by the tumour, possibly leading to urine retention, was thought to be the main reason for the high incidence of urinary tract infection among dogs with transmissible venereal tumours. Long-standing cases of transmissible venereal neoplasia had a higher chance of becoming bacteriuric compared with recent cases (OR=29.60). This study indicates that transmissible venereal tumour may he a predisposing factor for the development of urinary tract infection.Item Identification and typing of Pasteurella multocida: a review(Taylor and Francis, 2000-03) Blackall, P. J; Miflin, J. KFowl cholera is an important disease of both domesticated and wild birds ( Rimler & Glisson, 1997 ). As noted by Rimler & Glisson ( 1997 ), the disease is often classified into two forms, acute and chronic, despite the fact that field outbreaks often present clinical signs and lesions that are inter- mediate between these two forms. In the classic acute form, death may be the only indication of disease noted ( Rimler & Glisson, 1997). Astute observation of birds in the hours before death can reveal signs such as fever, anorexia, ruffled feath- ers, mouth discharge, diarrhoea and increased respiratory rate. In the chronic form, signs are typically limited to localized infections – swellingtaxa have been described. Table 1 shows the formally recognized species as well as the various un-named taxa that have been recorded as being present in birds. Hence, the identification of a Pasteurella-like isolate from an avian host is a challenging task – the complexity of which is not often appreciated by those who think of the genus Pasteurella as consisting of two to three species.Item Productivity of local chickens under village management Conditions(Kluwer Academic, 2002) Mwalusanya N. A; Katule A. M; Mutayoba S. K; Mtambo, M. M. A; Olsen J. E; Minga U. MThe productivity of local chickens under village management conditions was studied in six villages situated in three climatic zones within Morogoro District in Tanzania. Two villages were picked in each climatic zone (warm and wet, warm and dry, cool and wet) for the study. The data were obtained by actual measurement, qualitative observations and interview of members of the households directly responsible for the care of chickens. In addition, data sheets were given to selected farmers to record the performance of their chickens. The mean £ock size for the three zones was 16.2, with a range of 2 to 58. The overall mean clutch size, egg weight and hatchability were 11.8, 44.1 g and 83.6%, respectively. The overall mean chick survival rate to 10 weeks of age was 59.7%. The mean live weights for cocks and hens were 1948 g and 1348 g, respectively. The mean growth rates to the age of 10 weeks were 4.6 g/day and 5.4 g/day, while those from 10 to 14 weeks of age were 8.4 g/day and 10.2 g/day for female and male birds, respectively. The age at ¢rst lay ranged between 6 and 8 months, and the average hen had three laying cycles per year. Most of the chickens were left to scavenge during the day and were provided with simple housing at night (95.2% of the owners). Only small amounts of supplementary feeds were occasionally given and minimal health care was provided. It was concluded that the low productivity of chickens was partly due to the prevailing poor management practices, in particular the lack of proper health care, poor nutrition and housing.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 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 Porcine circovirus 2 uses heparan sulfate and chondroitin sulfate B glycosaminoglycans as receptors for its attachment to host cells(Journal of virology, 2006) Misinzo, Gerald; Delputte, Peter L; Meerts, Peter; Lefebvre, David J; Nauwynck, Hans JMonocyte/macrophage lineage cells are target cells in vivo for porcine circovirus 2 (PCV2) replication. The porcine monocytic cell line 3D4/31 supports PCV2 replication in vitro, and attachment and internalization kinetics of PCV2 have been established in these cells. However, PCV2 receptors remain unknown. Glycosami- noglycans (GAG) are used by several viruses as receptors. The present study examined the role of GAG in attachment and infection of PCV2. Heparin, heparan sulfate (HS), chondroitin sulfate B (CS-B), but not CS-A, and keratan sulfate reduced PCV2 infection when these GAG were incubated with PCV2 prior to and during inoculation of 3D4/31 cells. Enzymatic removal of HS and CS-B prior to PCV2 inoculation of 3D4/31 cells significantly reduced PCV2 infection. Similarly, when PCV2 virus-like particles (VLP) were allowed to bind onto 3D4/31 cells in the presence of heparin and CS-B, attachment was strongly reduced. Titration of field isolates and low- and high-passage laboratory strains of PCV2 in the presence of heparin significantly reduced PCV2 titers, showing that the capacity of PCV2 to bind GAG was not acquired during in vitro cultivation but is an intrinsic feature of wild-type virus. When Chinese hamster ovary (CHO) cells were inoculated with PCV2, relative percentages of PCV2-infected cells were 27% ⴞ 8% for HS-deficient and 12% ⴞ 10% for GAG-deficient cells compared to wild-type cells (100%). Furthermore, it was shown using heparin-Sepharose chromatography that both PCV2 and PCV2 VLP directly interacted with heparin. Together, these results show that HS and CS-B are attachment receptors for PCV2Item 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 New Leptospira serovar Sokoine of serogroup Icterohaemorrhagiae from cattle in Tanzania(Microbiology Society, 2006-04) Mgode, Georgies; Machang'u, Robert S; Goris, Marga; Engelberts, Mirjam F MThe prevalence of leptospirosis is generally high in domestic animals and rodents in Tanzania. Identification of Leptospira isolates from cattle was carried out to establish prevalent Leptospira serovars. Serological typing was done based on monoclonal antibodies and the standard cross-agglutination absorption test. Molecular typing involved pathogenic- and saprophytic-specific PCRs and a PCR specifically amplifying DNA from the species Leptospira kirschneri. DNA fingerprinting with primers derived from sequences of insertion elements IS1500 and IS1533 was carried out. Both serological and molecular characterization indicated that one of the Leptospira isolates, coded RM1, represents a new serovar of the species L. kirschneri of serogroup Icterohaemorrhagiae. The serovar name Sokoine is proposed for this new Leptospira isolate.Item Demographic and spatio-temporal variation in human plague at a persistent focus in Tanzania(Elsevier, 2006-10) Makundi, R.H; Davis, S; Machan"gu, R.S; Leirs, HHuman plague in the Western Usambara Mountains in Tanzania has been a public health problem since the first outbreak in 1980. The wildlife reservoir is unknown and eradication measures that have proved effective elsewhere in Tanzania appear to fail in this region. We use census data from 2002 and hospital records kept since 1986 to describe the temporal, spatial and demographic variation in human plague. A seasonal peak in cases occurs from December to February with the numbers of cases during this peak varying between 0 and 1150. Variation in incidence, calculated for each village as the mean number of cases per thousand inhabitants per year, indicates that human plague is concentrated around a group of three neighbouring, relatively isolated, high-altitude villages; Nywelo, Madala and Gologolo. However, there was no evidence that these villages were acting as a source of infection for the remainder of the focus. The likelihood of becoming infected with plague is highest between the ages of 5 and 19 and lowest for adult men. This was most clear in the ward encompassing the three high-incidence villages where the risk of plague among children aged 10–14 was 2.2 times higher than for adults aged 30–34, and among adults aged 30–34, the risk was 2.4 times higher for women than men.Item Multilocus sequence typing method for identification and genotypic classification of pathogenic Leptospira species(2006-11-23) Ahmed, N.; Devi, S. M.; Valverde, M.; Vijayachari, P.; Machang'u, R. S.; Ellis, W. A.; Hartskeerl, R. A.Leptospira are the parasitic bacterial organisms associated with a broad range of mammalian hosts and are responsible for severe cases of human Leptospirosis. The epidemiology of leptospirosis is complex and dynamic. Multiple serovars have been identified, each adapted to one or more animal hosts. Adaptation is a dynamic process that changes the spatial and temporal distribution of serovars and clinical manifestations in different hosts. Serotyping based on repertoire of surface antigens is an ambiguous and artificial system of classification of leptospiral agents. Molecular typing methods for the identification of pathogenic leptospires up to individual genome species level have been highly sought after since the decipherment of whole genome sequences. Only a few resources exist for microbial genotypic data based on individual techniques such as Multiple Locus Sequence Typing (MLST), but unfortunately no such databases are existent for leptospires. We for the first time report development of a robust MLST method for genotyping of Leptospira. Genotyping based on DNA sequence identity of 4 housekeeping genes and 2 candidate genes was analyzed in a set of 120 strains including 41 reference strains representing different geographical areas and from different sources. Of the six selected genes, adk, icdA and secY were significantly more variable whereas the LipL32 and LipL41 coding genes and the rrs2 gene were moderately variable. The phylogenetic tree clustered the isolates according to the genome-based species. The main advantages of MLST over other typing methods for leptospires include reproducibility, robustness, consistency and portability. The genetic relatedness of the leptospires can be better studied by the MLST approach and can be used for molecular epidemiological and evolutionary studies and population genetics.Item Survival of avirulent thermostable Newcastle disease virus (strain I-2) in raw, baked, oiled, and cooked white rice at ambient temperatures(2007) Wambura, Philemon Nyangi; Meers, Joanne; Spradbrow, PeterRaw white rice has not been considered a good carrier for oral vaccination, probably because of its antiviral activity. Methods are required to overcome antiviral activity in raw white rice. This study was carried out to determine the effects of various treatments of raw white rice on the survival of strain I-2 of Newcastle disease virus. These included cooking and baking the rice or mixing the rice with vegetable oil prior to coating with vaccine virus. The vaccine-coated rice was then stored for 30 min and 24 h, followed by quantitative recovery of the virus. Thirty min after mixing, uncooked, cooked, and baked rice, and rice mixed with vegetable oil showed titers of 10(6.2), 10(7.2), 10(6.6), and 10(7.0) EID(50)/0.1 ml, respectively. After storage for 24 h at 22-25oC, the titers dropped to 10(5.0), 10(6.5), 10(5.0), and 10(6.0) EID(50)/0.1 ml for uncooked, cooked, baked, and oiled rice, respectively.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 Occurrence of Trypanosoma in Nile tilapia in Lake Victoria, Kenya(2008) Kamundia, P.W.; Mbuthia, P.G.; Waruiru, R. M.; Njagi, L. W.; Nyaga, P. N.; Mdegela, R.H.; Byarugaba, D. K.; Otieno, R. O.Oreochromis niloticus (Nile tilapia) and Lates niloticus (Nile perch) are the most abundant species and economically important fish in Lake Victoria. The former are omnivores and the latter are carnivorous. The carrier status of haemoparasites in fish was investigated in 22 randomly selected fish specimens, 12 Oreochromis niloticus (Nile tilapia) and 10 Lates niloticus (Nile perch). Live fish were bought from Homabay on the shores of Lake Victoria in Kenya. All the fish were bled by cardiac puncture using a 22 gauge needle. Thin blood smears were made, stained with Giemsa and observed under a light microscope. About 42% of Nile tilapia and 0% Nile perch were found to be infected with Trypanosoma spp. Oreochromis niloticus seems to be more susceptible to Trypanosoma spp. infection than the L. niloticus.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 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 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 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 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 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.